HK1119722A

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Info

Publication number: HK1119722A
Application number: HK08112980.9A
Authority: HK
Inventors: 约瑟夫‧M.‧德西蒙; 贾森‧P.‧罗兰德; 金吉尔‧M.‧丹尼森
Original assignee: 北卡罗来纳大学查珀尔希尔分校
Priority date: 2004-02-13
Filing date: 2005-02-14
Publication date: 2009-03-13

Description

Functional materials and novel methods for manufacturing microfluidic devices

Cross Reference to Related Applications

This application is based on and claims priority from U.S. provisional patent application serial No.60/544,905 filed on 13/2/2004, which is incorporated herein by reference in its entirety.

Government rights and interests

The invention was made under U.S. government support from naval survey bureau No. n000140210185 and the STC project of the national science foundation under agreement No. che-9876674. The U.S. government has certain rights in this invention.

Technical Field

The presently disclosed subject matter relates to functional materials and their use for fabricating and utilizing micro-and nano-scale devices.

Abbreviations

AC-AC

Ar is argon

Degree centigrade

cm is equal to centimeter

8-CNVE-perfluoro (8-cyano-5-methyl-3, 6-dioxa-1-octene)

CSM ═ cure site monomer (cure site monomer)

CTFE ═ chlorotrifluoroethylene

g is g ═ g

h is hour

1-HPFP ═ 1, 2, 3, 3, 3-pentafluoropropene

2-HPFP ═ 1, 1, 3, 3, 3-pentafluoropropene

HFP ═ hexafluoropropene

HMDS (hexamethyldisilazane)

IL-imprint lithography

MCP (micro contact printing)

Me is methyl

MEA (membrane electrode assembly)

MEMS (micro-electromechanical system)

MeOH ═ methanol

MIMIC (micro-molded in capillary)

mL to mL

mm-mm

mmol ═ mmol

Mn is number average molecular weight

m.p. ═ melting point

mW-milliwatt

NCM (nano contact molding)

NIL (nano imprint lithography)

nm-nm

Pd ═ palladium

PAVE ═ perfluoro (alkyl vinyl) ether

PDMS ═ poly (dimethylsiloxane)

PEM (proton exchange Membrane)

PFPE ═ perfluoropolyether

PMVE-perfluoro (methyl vinyl) ether

PPVE ═ perfluoro (propyl vinyl) ether

PSEPVE ═ perfluoro-2- (2-fluorosulfonylethoxy) propyl vinyl ether

PTFE ═ polytetrafluoroethylene

SAMIM (solvent assisted micromolding)

SEM (scanning electron microscopy)

Si ═ silicon

TFE ═ tetrafluoroethylene

Mum ═ micron

UV (ultraviolet) ray

W ═ watt

ZDOL ═ poly (oxytetrafluoroethylene-co-oxydifluoromethylene) alpha, omega-diols

(poly(tetrafluoroethylene oxide-co-difluoromethylene oxide)α，ω-diol)

Background

Microfluidic devices that were open in the early nineties of the twentieth century were fabricated from hard materials such as silicon and glass by using photolithography and etching techniques. See, Ouellette, J., the Industrial Physicist 2003, 8/9 months, 14-17; scherer, A. et al, Science 2000, 290, 1536-. However, photolithography and etching techniques are costly and labor intensive, require clean room conditions, and have some drawbacks from a material standpoint. For this reason, soft materials have emerged as an alternative material for microfluidic device fabrication. The use of soft materials enables the manufacture and actuation of devices containing valves, pumps and mixers. See, e.g., Ouellette, J., The Industrial Physicist 2003, August/September, 14-17; scherer, A., et al., Science 2000, 290, 1536-; unger, m.a., et al, Science 2000, 288, 113-; McDonald, j.c., et al, acc.chem.res.2002, 35, 491-. For example, one such microfluidic device can control the direction of overflow without the use of mechanical valves. See Zhao, b. et al, Science 2001, 291, 1023-.

The increasing complexity of microfluidic devices has created a demand for such devices for a rapidly growing number of applications. To this end, the use of flexible materials has prompted the development of microfluidics into useful technologies that have found applications in genomic mapping, rapid separation, sensors, nanoreactions, ink-jet printing, drug delivery, Lab-on-a-Chip (Lab-on-a-Chip), in vitro diagnostics, injection nozzles, biological research, and drug screening. See, e.g., Ouellette, J., The Industrial Physicist 2003, August/September, 14-17; scherer, A., et al, Science 2000, 290, 1536-; unger, m.a., et al., Science 2000, 288, 113-; McDonald, j.c., et al, acc.chem.res.2002, 35, 491-; thorsen, T., et al, Science 2002, 298, 580-.

Poly (dimethylsiloxane) (PDMS) is the preferred flexible material for many microfluidic device applications. See Scherer.A., et al, Science 2000, 290, 1536-; unger, m.a., et al., Science 2000, 288, 113-; McDonald, j.c., et al, acc.chem.res, 2002, 35, 491-; thorsen, T, et al, Science 2002, 298, 580-584 and Liu, J Et al, anal chem.2003, 75, 4718-. PDMS materials offer many attractive properties in microfluidic applications. After crosslinking, PDMS becomes an elastomeric material with a low young's modulus, for example, about 750 kPa. See Unger, m.a., et al, Science2000, 288, 113-. This property allows PDMS to conform to the surface and form a reversible seal. In addition, PDMS has a low surface energy, e.g., about 20erg/cm²This facilitates release from the mold after patterning. See Scher.A. et al, Science2000, 290, 1536-1539; McDonald, J.C., et al, Acc.chem.Res.2002, 35, 491-.

Another important feature of PDMS is its outstanding gas permeability. This property allows bubbles within the channels of the microfluidic device to permeate out of the device. This property is also useful for maintaining cells and microorganisms within structural features of the microfluidic device. The non-toxic nature of polysiloxanes such as PDMS is also beneficial in this respect and has potential applications in the field of medical implants. McDonald.J.C. et al Acc.chem.Res.2002, 35, 491-.

Many current PDMS microfluidic devices are based on184(DoWCorning, Midland, Mich.) of Mich. 184 is thermal cure by platinum catalyzed hydrosilation.184 may take up to five hours to fully cure. However, it has recently been reported to have properties comparable to those used for soft lithography184 mechanical properties similar to those of a photocurable PDMS material. See Choi.K.M. et al, J.Am.chem.Soc.2003, 125, 4060-.

Despite the above advantages, PDMS has the disadvantage in microfluidic applications that it swells in most organic solvents. Therefore, PDMS-based microfluidic devices have limited compatibility with various organic solvents. See Lee, J.N. et al, anal. chem.2003, 75, 6544-. Organic solvents that swell PDMS are hexane, ether, toluene, dichloromethane, acetone and acetonitrile. See Lee, J.N. et al, anal. chem.2003, 75, 6544-. Swelling of a PDMS microfluidic device by an organic solvent may interfere with its microscale structural features, such as a channel or channels, and may restrict or completely shut off the flow of organic solvent through the channel. Thus, microfluidic applications for PDMS-type devices are limited to the use of fluids such as water that do not swell PDMS. As a result, those applications requiring the use of organic solvents may require the use of microfluidic systems fabricated from hard materials such as glass and silicon. See Lee, J.N. et al, anal. chem.2003, 75, 6544-. However, this solution is limited by the disadvantages of microfluidic devices made of hard materials.

Additionally, PDMS-type devices and materials are not inert enough to make them useful even in aqueous chemical processes. For example, PDMS reacts readily with weak and strong acids and bases. PDMS type devices are also known to contain extractables, especially extractable oligomers and cyclic siloxanes, especially after exposure to acids and bases. Since PDMS is easily swelled by organic substances, hydrophobic materials, even those slightly soluble in water, partition into the PDMS-type material used to construct PDMS-type microfluidic devices.

Thus, elastomeric materials that exhibit the attractive mechanical properties of PDMS, combined with swelling resistance in common organic solvents, would extend the application of microfluidic devices to a variety of new chemical applications not achievable by current PDMS-based devices. Thus, the approach described by the presently disclosed subject matter uses elastomeric materials, more specifically functionalized perfluoropolyether (PFPE) materials, that resist swelling in common organic solvents, thereby making microfluidic devices possible.

The functionalized PFPE materials are liquid at room temperature, exhibit low surface energy, low modulus, high gas permeability, and low toxicity, and are increasingly characterized by extreme chemical resistance. See Scheirs, j., Modem Fluoropolymers; john Wiley & Sons, Ltd.: new york, 1997; pp 435-. Furthermore, PFPE materials exhibit hydrophobicity and lyophobicity. Thus, PFPE materials are often used as lubricants on high performance machines operating under harsh conditions. The synthesis and solubility of PFPE materials in supercritical carbon dioxide has been reported. See Bunyard, w, et al, Macromolecules 1999, 32, 8224-. In addition to PFPE, fluoroelastomers may also include fluoroolefin-based materials, including, but not limited to, copolymers of tetrafluoroethylene, hexafluoropropylene, vinylidene fluoride, and alkyl vinyl ethers, often with the addition of additional cure site monomers for crosslinking.

PFPE microfluidic devices have been previously reported by Rolland, J.et al, JACS 2004, 126, 2322-2323. The device is made from a functionalized PFPE material (e.g., PFPE dimethacrylate (MW 4,000g/mol)) having a viscosity of about 800 cSt. The material is end-capped-functionalized with free-radically polymerizable methacrylate groups and is free-radically UV light cured with a photoinitiator. In the above by Rolland, j. et al, by creating a multilayer PFPE device using a specific partial UV curing technique, the adhesion is weak and generally not strong enough for many applications. Furthermore, the bonding techniques described by Roland, j.

The presently disclosed subject matter describes the use of fluoroelastomers, particularly functionalized perfluoropolyethers, as materials for the fabrication of solvent resistant micro-and nano-scale structures such as microfluidic devices. The use of fluoroelastomers and functionalized perfluoropolyethers, particularly as materials for the manufacture of microfluidic devices, solves the problem of swelling in organic solvents exhibited by microfluidic devices made from other polymeric materials such as PDMS. Thus, PFPE-based microfluidic devices can be used to control the flow of small volumes of fluids, such as organic solvents, and can be used to perform micro-scale and nano-scale chemical reactions that are not suitable for other polymeric microfluidic devices.

Disclosure of Invention

The presently disclosed subject matter Provides Functionalized Perfluoropolyether (PFPE) materials for use in the manufacture of microfluidic devices. In some embodiments, the presently disclosed subject matter provides methods of bonding two-dimensional and three-dimensional micro-and/or nano-scale structures, such as microfluidic networks, to substrates. Furthermore, in some embodiments, the presently disclosed subject matter provides methods of forming hybrid microfluidic devices (e.g., microfluidic devices comprising a perfluoropolyether layer bonded to a second polymer layer, wherein the second polymer layer comprises, for example, a poly (dimethylsiloxane) layer).

The presently disclosed subject matter also provides methods of fabricating micro-scale and/or nano-scale structures, such as microfluidic devices, by using sacrificial layers of degradable materials. More specifically, the presently disclosed subject matter provides methods for fabricating micro-scale and/or nano-scale structures by using degradable or selectively soluble polymers as scaffolds (scaffold) for the production of complex, two-dimensional (2-D) and three-dimensional (3-D) microfluidic networks.

In addition, the presently disclosed subject matter provides functionalized materials for attaching biological and other "switchable" molecules to the interior surfaces of microfluidic channels. For example, attaching biomolecules, such as biopolymers, to the interior surfaces of microfluidic channels can ensure combinatorial peptide synthesis and/or rapid screening of enzyme-protein interactions. Furthermore, lining the microfluidic channels with catalyst may allow for rapid catalyst screening. Also, the introduction of switchable organic molecules into microfluidic channels may enable the fabrication of microfluidic devices that include hydrophilic channels and hydrophobic channels.

In some embodiments, the presently disclosed subject matter provides methods of using functionalized perfluoropolyether networks as gas separation membranes.

It is therefore an object of the presently disclosed subject matter to provide functionalized perfluoropolyether materials for the manufacture and utilization of micro-and nano-scale devices, including microfluidic devices. This and other objects are achieved in whole or in part by the presently disclosed subject matter.

The above described objects, other aspects and objects of the presently disclosed subject matter will become apparent from the description and the accompanying drawings, when taken in conjunction with the accompanying drawings and the embodiments described in detail below.

Drawings

Fig. 1A-1C are a series of schematic end views depicting the formation of a patterned layer of polymeric material in accordance with the presently disclosed subject matter.

Fig. 2A-2D are a series of schematic end views depicting a microfluidic device forming a patterned layer comprising two polymeric materials according to the presently disclosed subject matter.

Figures 3A-3C are schematic illustrations of embodiments of the presently disclosed methods for bonding a functionalized microfluidic device to a treated substrate.

Fig. 4A-4C are schematic diagrams of embodiments of the presently disclosed methods for fabricating multilayer microfluidic devices.

Fig. 5A and 5B are schematic illustrations of an embodiment of the presently disclosed method for functionalizing the interior surfaces of a microfluidic channel and the surface of a microtiter tube.

Fig. 5A is a schematic illustration of an embodiment of the presently disclosed method for functionalizing an interior surface of a microfluidic channel.

FIG. 5B is a schematic illustration of an embodiment of the presently disclosed method for functionalizing a surface of a microtiter tube.

Fig. 6A-6D are schematic illustrations of embodiments of the presently disclosed methods of fabricating micro-scale structures using degradable and/or selectively soluble materials.

Figures 7A-7C are schematic illustrations of embodiments of the presently disclosed method of fabricating complex structures in micro-scale and/or nano-scale devices using degradable and/or selectively soluble materials.

Fig. 8 is a schematic plan view of a microfluidic device according to the presently disclosed subject matter.

FIG. 9 is a schematic diagram of an integrated microfluidic system for biopolymer synthesis.

Fig. 10 is a schematic diagram of a system for flowing a solution or performing a chemical reaction in a microfluidic device according to the presently disclosed subject matter. The microfluidic device 800 is depicted in the schematic plan view shown in fig. 8.

Detailed Description

The presently disclosed subject matter provides materials and methods for forming microfluidic devices and for imparting chemical functionality to microfluidic devices. In some embodiments, the presently disclosed methods include introducing chemical functional groups that promote and/or enhance adhesion between various layers of the microfluidic device. In some embodiments, the chemical functional group promotes and/or enhances adhesion between a layer of the microfluidic device and another surface. Thus, in some embodiments, the presently disclosed subject matter provides methods of bonding two-dimensional and three-dimensional microfluidic networks to substrates. In some embodiments, the presently disclosed methods can be used to bond perfluoropolyether (PFPE) materials to other materials such as poly (dimethylsiloxane) (PDMS) materials, polyurethane materials, polysiloxane-containing polyurethane materials, and PFPE-PDMS block copolymer materials. Thus, in some embodiments, the presently disclosed subject matter provides methods of forming hybrid microfluidic devices (e.g., microfluidic devices comprising a perfluoropolyether layer bonded to a polydimethylsiloxane layer, a polyurethane layer, a polysiloxane-containing polyurethane layer, and a PFPE-PDMS block copolymer layer).

In some embodiments, the method comprises introducing chemical functional groups to an inner surface of a microfluidic channel and/or a microtiter tube. In some embodiments, the introduction of chemical functional groups on the inner surface of the microfluidic channel and/or the microtiter tube provides for the attachment of biopolymers and other small organic "switchable" molecules that can affect the hydrophobicity or reactivity of the microfluidic channel and/or the microtiter tube.

In some embodiments, the presently disclosed subject matter provides methods of forming micro-scale and/or nano-scale structures in which a scaffold of degradable or selectively soluble polymers is used to form channels, for example, inside a microfluidic device. Thus, the molding methods disclosed herein can form complex three-dimensional networks of microfluidic channels in a one-step process.

The presently disclosed subject matter is now described more fully with reference to the accompanying drawings and examples, in which representative embodiments are shown. The presently disclosed subject matter, however, may be embodied in different forms and should not be construed as limited to the embodiments set forth herein. These embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of these embodiments to those skilled in the art.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the presently described subject matter belongs. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

Throughout the specification and claims, a given chemical formula or name shall include all optical and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

Definition of

The term "microfluidic device" as used herein generally refers to a device through which a material, particularly a fluid-containing material (e.g., a liquid), can be transported, in some embodiments on the micron scale, and in some embodiments on the nanometer scale. Thus, the microfluidic devices described by the presently disclosed subject matter can include micro-scale structural features, nano-scale structural features, and combinations thereof.

Accordingly, microfluidic devices typically include structured or functionalized features on the order of millimeters or less in size that are capable of manipulating fluids at flow rates on the order of microliters/min or less. Typically, such features include, but are not limited to, channels, fluid reservoirs, reaction chambers, mixing chambers, and separation zones. In some embodiments, the channel comprises at least one cross-sectional dimension in a range from about 0.1 μm to about 500 μm. The use of dimensions on this order of magnitude may allow the introduction of a greater number of channels in a smaller area and the use of a smaller volume of fluid.

The microfluidic device can exist alone or can be part of a microfluidic system, including but not limited to: introducing fluids, e.g., samples, reagents, buffers, etc., into and/or through the system; a detection device or system; a reagent, product or data storage system; and a control system for controlling fluid transport and/or direction within the apparatus, monitoring and controlling environmental conditions such as temperature, current, etc. to which the fluid in the apparatus is subjected.

The term "device" as used herein includes, but is not limited to, microfluidic devices, microtiter plates, tubes, hoses, and the like.

As used herein, the terms "channel," "microscale channel," and "microfluidic channel" are used interchangeably and refer to a pocket or cavity formed in a material by transferring a pattern from a patterned substrate into the material or by any suitable material removal technique, or to a pocket or cavity in combination with any suitable fluid-conducting structure, such as a tube, capillary, or similar structure, disposed in the pocket or cavity.

As used herein, the terms "flow channel" and "control channel" are used interchangeably and can refer to a channel in a microfluidic device through which a material, such as a fluid (e.g., a gas or a liquid), can flow. More specifically, the term "flow channel" refers to a channel through which the material, such as a solvent or chemical agent, can flow. Furthermore, the term "control channel" refers to a flow channel through which a material, such as a fluid (e.g., a gas or a liquid), can flow by operating a valve or a pump.

The term "valve" as used herein, unless otherwise specified, refers to a configuration in which two channels are separated by an elastomeric segment, such as a PFPE segment, that is capable of deflecting into or out of one of the channels, such as a flow channel, in response to a driving force exerted on the other channel, such as a control channel. The term "valve" also includes one-way valves, which comprise channels separated by beads.

The term "pattern" as used herein refers to a channel or microfluidic channel, or an integrated network of microfluidic channels, which in some embodiments are capable of crossing at a predetermined point. The pattern can also include one or more of microscale or nanoscale fluid reservoirs, microscale or nanoscale reaction chambers, microscale or nanoscale mixing chambers, and microscale or nanoscale separation regions.

The term "intersect" as used herein means meeting at a point, meeting at a point and crossing or spanning, or meeting at a point and merging overlapping. More specifically, the term "crossover" as used herein describes an embodiment in which two channels meet at a point, meet at a point and intersect or span each other, or meet at a point and overlap each other. Thus, in some embodiments, two channels can intersect, i.e., meet at one point or meet at one point and intersect each other, as well as be in fluid communication with each other. In some embodiments, the two channels can intersect, i.e., meet and overlap each other at a point, and do not achieve fluid communication with each other, as is the case when the flow channel and the control channel intersect.

The term "communicate" and grammatical variations thereof as used herein (e.g., a first component "communicates with" or "communicates with" a second component ") are intended to refer to a structural, functional, mechanical, electrical, optical, or fluidic relationship between two or more components or elements, or any combination thereof. Likewise, the fact that one component is said to be in communication with a second component is not intended to exclude the possibility that additional components may be present between and/or operatively associated with or interfitted with the first and second components.

The terms "in," "on," "into," "onto," and "across" devices generally have equivalent meanings when referring to the use of a microfluidic device for handling containment or movement of fluids.

The term "monolithic" as used herein refers to a structure that includes or functions as a single, uniform structure.

The term "non-biological organic material" as used herein refers to organic materials other than biological materials, i.e., those compounds having covalent carbon-carbon bonds. The term "biomaterial" as used herein includes nucleic acid polymers (e.g., DNA, RNA), amino acid polymers (e.g., enzymes, proteins, etc.) and small organic compounds (e.g., steroids, hormones), wherein the small organic compounds are biologically active, particularly for humans or animals of commercial interest such as pets and livestock, and wherein the small organic compounds are used primarily for therapeutic or diagnostic purposes. Although biomaterials are advantageous for pharmaceutical and biotechnological applications, a large number of applications involve chemical processes enhanced by those other than biomaterials, i.e. non-biological organic materials.

The term "partially cured" as used herein refers to a process wherein less than about 100% of the polymerizable groups are reacted. Thus, the term "partially cured material" refers to a material that undergoes a partial curing process.

The term "fully cured" as used herein refers to a process wherein about 100% of the polymerizable groups are reacted. Thus, the term "fully cured material" refers to a material that undergoes a full curing process.

The terms "a," "an," and "the" when used in this application, including the claims, mean "one or more" in accordance with established patent statutory convention. Thus, for example, reference to a "microfluidic channel" includes a plurality of such microfluidic channels, and so forth.

II. Material

The presently disclosed subject matter broadly describes and employs solvent resistant, low surface energy polymeric materials, particularly materials formed by casting a liquid PFPE precursor material onto a patterned substrate and then curing the liquid PFPE precursor material to produce a patterned layer of functionalized PFPE material, which can be used to form microfluidic devices.

Representative solvent resistant elastomeric-based materials include, but are not limited to, fluorinated elastomeric-type materials. The term "solvent resistant" as used herein refers to an elastomeric material that neither swells nor dissolves in common hydrocarbon organic solvents or acidic or basic aqueous solutions. Representative fluorinated elastomer-type materials include, but are not limited to, perfluoropolyether (PFPE) -based materials.

The functionalized liquid PFPE materials exhibit desirable properties to enable their use in microfluidic devices. For example, functionalized PFPE materials typically have low surface energies (e.g., about 12 mN/m); is non-toxic, UV and visible transparent, and is highly gas permeable; and cured into a tough, durable, highly fluorinated elastomeric or glassy material having excellent release (realease) properties and resistance to swelling. The properties of these materials can be adjusted within wide limits by suitable selection of additives, fillers, reactive comonomers and functionalizing agents. Such properties that it is desirable to alter include, but are not limited to, modulus, tear strength, surface energy, permeability, functionality, mode of cure, solubility and swellability, and the like. The non-swelling properties and easy release properties of the presently disclosed PFPE materials allow microfluidic devices to be fabricated.

A perfluoropolyether material prepared from a liquid PFPE precursor material having a viscosity of less than about 100 centistokes.

Those skilled in the art will recognize that perfluoropolyethers (PFPEs) have been used in many applications over the past 25 years. Commercial PFPE materials are prepared by polymerization of perfluorinated monomers. The first member of this type is prepared by cesium fluoride catalyzed polymerization of hexafluoropropylene oxide (HFPO), which is designated as (DuPont, Wilmington, Delaware, USA) is a series of branched polymers. Similar polymers are produced by UV catalyzed photo-oxidation of hexafluoropropylene: (Y) (Solvay Solexis, Brussels, Belgium). Furthermore, a linear polymer (CZ) (Solvay) was prepared by a similar method, but using tetrafluoroethylene. Finally, a fourth polymer () (Daikin Industries, Ltd., Osaka, Japan) is produced by polymerization of tetrafluorooxetanes followed by direct fluorination. The structures of these fluids are listed in table I. TABLE II Properties of some members of lubricant containing PFPE typeThe data can be obtained. Likewise, the physical properties of the functionalized PFPE are provided in table III. In addition to these commercially available PFPE fluids, a new series of structures were prepared by direct fluorination techniques. Representative structures of these new PFPE materials are given in table IV. Of the above PFPE fluids, onlyAndz has been widely used in these applications. See Jones, W.R., Jr., The Properties of fluorinated thermal used for Space Applications, NASA Technical Memorandum 106275(July 1993), The entire contents of which are incorporated herein by reference. Thus, applications of such PFPE materials are provided in the presently disclosed subject matter.

Table i name and chemical structure of commercially available PFPE fluids

PFPE physical Properties

TABLE III PFPE physical Properties of functionalized PFPE

Table iv. name and chemical structure of representative PFPE fluids

Name (R)	Structure of the product^a
Name (R)	Structure of the product^a	Perfluoropoly (oxymethylene) (PMO)	CF₃O(CF₂O)_xCF₃
Perfluoropoly (ethylene oxide) (PEO)	CF₃O(CF₂CF₂O)_xCF₃	Perfluoropoly (oxymethylene) (PMO)	CF₃O(CF₂O)_xCF₃
Perfluoropoly (ethylene oxide) (PEO)	CF₃O(CF₂CF₂O)_xCF₃	Perfluoropoly (dioxolane) (DIOX)	CF₃O(CF₂CF₂OCF₂O)_xCF₃
Perfluoropoly (trioxane ring) (TRIOX)	CF₃O[(CF₂CF₂O)₂CF₂O]_xCF₃	Perfluoropoly (dioxolane) (DIOX)	CF₃O(CF₂CF₂OCF₂O)_xCF₃

^aWhere x is any integer.

In some embodiments, the perfluoropolyether precursor includes a poly (oxytetrafluoroethylene-co-oxydifluoromethylene) alpha, omega-diol that, in some embodiments, is capable of photocuring to form one of a perfluoropolyether dimethacrylate and a perfluoropolyether stilbene compound. A representative scheme for the synthesis and photocuring of functionalized perfluoropolyethers is provided in scheme 1.

Scheme 1 Synthesis and photocuring of functionalized perfluoropolyethers

Perfluoropolyether materials prepared from liquid PFPE precursor materials having a viscosity greater than about 100 centistokes.

These methods are provided below to promote and/or increase adhesion between a layer of PFPE material and another material and/or substrate and to add chemical functional groups to a surface comprising the PFPE material having properties selected from the group consisting of: a viscosity of greater than about 100 centistokes (cSt); and a viscosity of less than about 100cSt, with the proviso that the liquid PFPE precursor material having a viscosity of less than 100cSt is not a free radical photo-curable PFPE material. As provided herein, the viscosity of a liquid PFPE precursor material refers to the viscosity of the material prior to functionalization (e.g., with methacrylate or styrene groups).

Thus, in some embodiments, the PFPE material is prepared from a liquid PFPE precursor material having a viscosity greater than about 100 centistokes (cSt). In some embodiments, the liquid PFPE precursor is terminated with a polymerizable group. In some embodiments, the polymerizable group is selected from acrylate, methacrylate, epoxy, amino, carboxyl, anhydride, maleimide, isocyanate, alkene, and styrenic groups.

In some embodiments, the perfluoropolyether material includes a backbone structure selected from the group consisting of:

wherein x is present and absent and includes a capping group when present, and n is an integer from 1 to 100.

In some embodiments, the PFPE liquid precursor is synthesized from hexafluoropropylene oxide, as shown in scheme 2.

Scheme 2 Synthesis of liquid PFPE precursor Material from Hexafluoropropane oxide

In some embodiments, the liquid PFPE precursor is synthesized from hexafluoropropylene oxide, as shown in scheme 3.

Scheme 3 Synthesis of liquid PFPE precursor Material from Hexafluoropropylene oxide

In some embodiments, the liquid PFPE precursor comprises a material that is chain extended such that two or more chains are linked together prior to addition of the polymerizable group. Thus, in some embodiments, a "linker group" links two chains together to form one molecule. In some embodiments, the linker group links three or more chains together as shown in scheme 4.

Scheme 4 linker groups linking together 3 PFPE chains

In some embodiments, X is selected from the group consisting of isocyanate, acid chloride, epoxy and halogen. In some embodiments, R is selected from the group consisting of acrylate, methacrylate, styrene, epoxy, carboxyl, anhydride, maleimide, isocyanate, olefin, and amine. In some embodiments, the circle represents any multifunctional molecule. In some embodiments, the multifunctional molecule comprises a cyclic molecule. PFPE refers to any of the PFPE materials provided above.

In some embodiments, the liquid PFPE precursor comprises a highly branched polymer provided in scheme 5, wherein PFPE refers to any of the PFPE materials provided above.

Scheme 5 highly branched PFPE liquid precursor Material

In some embodiments, the liquid PFPE material comprises an end-functionalized material selected from the group consisting of:

and

in some embodiments, the PFPE liquid precursor is terminated with an epoxy moiety, which can be photocured using a photoacid generator (photoacid generator). Photoacid generators suitable for use in the presently disclosed subject matter include, but are not limited to: bis (4-tert-butylphenyl) iodonium p-toluenesulfonate, bis (4-tert-butylphenyl) iodonium trifluoromethanesulfonate (triflate), (4-bromophenyl) diphenylsulfonium trifluoromethanesulfonate, (tert-butoxycarbonylmethoxynaphthyl) -diphenylsulfonium trifluoromethanesulfonate, (tert-butoxycarbonylmethoxyphenyl) diphenylsulfonium trifluoromethanesulfonate, (4-tert-butylphenyl) diphenylsulfonium trifluoromethanesulfonate, (4-chlorophenyl) diphenylsulfonium trifluoromethanesulfonate, diphenyliodonium-9, 10-dimethoxyanthracene-2-sulfonate, diphenyliodonium hexafluorophosphate, diphenyliodonium nitrate, diphenyliodonium perfluoro-1-butanesulfonate, diphenyliodonium p-toluenesulfonate, diphenyliodonium trifluoromethanesulfonate, (4-fluorophenyl) diphenylsulfonium trifluoromethanesulfonate, N-hydroxynaphthalimide (N-hydroxyynaphtalimide) trifluoromethanesulfonate, N-hydroxy-5-norbornene-2, 3-dicarboximide perfluoro-1-butanesulfonate, N-hydroxyphthalimide trifluoromethanesulfonate, [4- [ (2-hydroxytetradecyl) oxy ] phenyl ] phenyliodonium hexafluoroantimonate, (4-iodophenyl) diphenylsulfonium trifluoromethanesulfonate, (4-methoxyphenyl) diphenylsulfonium trifluoromethanesulfonate, 2- (4-methoxystyryl) -4, 6-bis (trichloromethyl) -1, 3, 5-triazine, (4-methylphenyl) diphenylsulfonium trifluoromethanesulfonate, (4-methylthiophenyl) methylphenylsulfonium trifluoromethanesulfonate, 2-naphthyldiphenylsulfonium trifluoromethanesulfonate, (4-phenoxyphenyl) diphenylsulfonium trifluoromethanesulfonate, (4-phenylthiophenyl) diphenylsulfonium trifluoromethanesulfonate, thiobis (triphenylsulfonium hexafluorophosphate), triarylsulfonium hexafluoroantimonate, triarylsulfonium hexafluorophosphate, triphenylsulfonium perfluoro-1-butanesulfonate, triphenylsulfonium trifluoromethanesulfonate, tris (4-tert-butylphenyl) sulfonium perfluoro-1-butanesulfonate, and tris (4-tert-butylphenyl) sulfonium trifluoromethanesulfonate.

In some embodiments, the liquid PFPE precursor cures into a highly UV and/or highly visible transparent elastomer. In some embodiments, the liquid PFPE precursor cures to an elastomer that is highly permeable to oxygen, carbon dioxide, and nitrogen, which permeability is a property that can promote the viability of biological fluids/cells located therein. In some embodiments, additives are added or various layers are formed to enhance the barrier properties of the device to molecules such as oxygen, carbon dioxide, nitrogen, dyes, agents, and the like.

In some embodiments, materials suitable for use with the presently disclosed subject matter include a polysiloxane material comprising fluoroalkyl functionalized Polydimethylsiloxane (PDMS) having the structure:

wherein:

r is selected from the group consisting of acrylate, methacrylate, and vinyl;

R_fcomprising fluoroalkyl chains; and

n is an integer from 1 to 100,000.

In some embodiments, materials suitable for use with the presently disclosed subject matter include styrenic materials comprising fluorinated styrene monomers selected from the group consisting of:

and

wherein R is_fIncluding fluoroalkyl chains.

In some embodiments, materials suitable for use with the presently disclosed subject matter include acrylic based materials including fluorinated acrylates or fluorinated methacrylates having the following structures:

Wherein:

r is selected from the group consisting of H, alkyl, substituted alkyl, aryl, and substituted aryl; and

R_fcomprising having a-CH between the perfluoroalkyl chain and the ester linkage₂-or-CH₂-CH₂Fluorine of spacerAn alkyl chain. In some embodiments, the perfluorinated alkyl group has a hydrogen substituent.

In some embodiments, materials suitable for use with the presently disclosed subject matter include triazine fluoropolymers containing fluorinated monomers.

In some embodiments, the fluorinated monomer or fluorinated oligomer capable of being polymerized or crosslinked by metathesis polymerization includes a functionalized olefin. In some embodiments, the functionalized olefin comprises a functionalized cyclic olefin.

II.C. fluoroolefin-based materials

Further, in some embodiments, the materials used herein are selected from highly fluorinated fluoroelastomers, such as those comprising at least 58 wt% fluorine as described in U.S. Pat. No.6,512,063 (issued to Tang), the entire contents of which are incorporated herein by reference. Such fluoroelastomers can be partially fluorinated or perfluorinated and contain from 25 to 70 weight percent (based on the weight of the fluoroelastomer) of a first monomer such as Vinylidene Fluoride (VF)₂) Or copolymerized units of Tetrafluoroethylene (TFE). The remaining units of the fluoroelastomer include one or more additional copolymerizable monomers, different from the first monomer, and selected from the group consisting of fluorine-containing olefins, fluorine-containing vinyl ethers, olefins, and combinations thereof.

These fluoroelastomers includePolymers of the DuPont Dow Elastomers, Wilmington, Delaware U.S.A.) and Kel-F types have been described for microfluidic applications in U.S. Pat. No.6,408,878 to Unger et al. However, these commercially available polymers have Mooney (Mooney) viscosities of from about 40 to 65(ML1+10, 121 ℃) such that they have a tacky, gel-like tack. When cured, they become hard, opaque solids. Are currently availableAnd Kel-F for microMeter-scale molding has limited utility. There is a need in the field of applications described herein for curable substances of similar composition but with lower viscosity and higher optical clarity. For lower viscosities at 20 ℃ (e.g., 2-32(ML1+10 at 121 ℃)) or more preferably as low as 80-2000cSt, the composition gives a pourable liquid, achieving more efficient curing.

More specifically, fluorine-containing olefins include, but are not limited to, vinylidene fluoride, Hexafluoropropylene (HFP), Tetrafluoroethylene (TFE), 1, 2, 3, 3, 3-pentafluoropropene (1-HPFP), Chlorotrifluoroethylene (CTFE), and vinyl fluoride.

Fluorine-containing vinyl ethers include, but are not limited to, perfluoro (alkyl vinyl) ethers (PAVE). More specifically, perfluoro (alkyl vinyl) ethers useful as monomers include perfluoro (alkyl vinyl) ethers of the general formula:

CF₂＝CFO(R_fO)_n(R_fO)_mR_f

Wherein each R is_fIndependently linear or branched C₁-C₆A perfluoroalkylene group, and m and n are each independently an integer from 0 to 10.

In some embodiments, the perfluoro (alkyl vinyl) ether comprises a monomer of the formula:

CF₂＝CFO(CF₂CFXO)_nR_f

wherein X is F or CF₃N is an integer of 0 to 5, and R_fIs linear or branched C₁-C₆A perfluoroalkylene group. In some embodiments, n is 0 or 1 and R_fIncluding 1 to 3 carbon atoms. Representative examples of such perfluoro (alkyl vinyl) ethers include perfluoro (methyl vinyl) ether (PMVE) and perfluoro (propyl vinyl) ether (PPVE).

In some embodiments, the perfluoro (alkyl vinyl) ether comprises a monomer of the general formula:

CF₂＝CFO[(CF₂)_mCF₂CFZO]_nR_f

wherein R is_fIs a perfluorinated alkyl group having 1 to 6 carbon atoms, m is 0 or an integer of 1, n is an integer from 0 to 5, and Z is F or CF₃. In some embodiments, R_fIs C₃F₇M is 0, and n is 1.

In some embodiments, the perfluoro (alkyl vinyl) ether monomer comprises a compound of the general formula:

CF₂＝CFO[(CF₂CF{CF₃}O)_n(CF₂CF₂CF₂O)_m(CF₂)_p]C_xF_2x+1

wherein m and n are each independently an integer from 0 to 10, p is an integer from 0 to 3, and x is an integer from 1 to 5. In some embodiments, n is 0 or 1, m is 0 or 1, and x is 1.

Other examples of useful perfluoro (alkyl vinyl ethers) include:

CF₂＝CFOCF₂CF(CF₃)O(CF₂O)_mC_nF_2n+1

wherein n is an integer from 1 to 5, and m is an integer from 1 to 3. In some embodiments, n is 1.

In embodiments where copolymerized units of perfluoro (alkyl vinyl) ether (PAVE) are present in the presently described fluoroelastomers, the PAVE content is generally from 25 to 75 weight percent, based on the total weight of the fluoroelastomer. If PAVE is perfluoro (methyl vinyl) ether (PMVE), the fluoroelastomer contains 30-55 weight percent copolymerized PMVE units.

Olefins that may be used in the presently described fluoroelastomers include, but are not limited to, ethylene (E) and propylene (P). In embodiments wherein copolymerized units of an olefin are present in the presently described fluoroelastomers, the olefin content is generally from 4 to 30 weight percent.

Furthermore, in some embodiments, the presently described fluoroelastomers can include units of one or more cure site monomers. Examples of suitable cure site monomers include: i) a bromine-containing olefin; ii) an iodine-containing olefin; iii) a bromine-containing vinyl ether; iv) iodine-containing vinyl ethers; v) a fluorine-containing olefin having a nitrile group; vi) a fluorine-containing vinyl ether having a nitrile group; vii)1, 1, 3, 3, 3-pentafluoropropene (2-HPFP); viii) perfluoro (2-phenoxypropyl vinyl) ether; and ix) a non-conjugated diene.

The brominated cure site monomer can contain other halogens, preferably fluorine. An example of a brominated olefin cure site monomer is CF₂＝CFOCF₂CF₂CF₂OCF₂CF₂Br; bromotrifluoroethylene; 4-bromo-3, 3, 4, 4-tetrafluorobutene-1 (BTFB); and other monomers such as vinyl bromide, 1-bromo-2, 2-difluoroethylene; perfluoroallyl bromide; 4-bromo-1, 1, 2-trifluorobutene-1; 4-bromo-1, 1, 3, 3, 4, 4-hexafluorobutene; 4-bromo-3-chloro-1, 1, 3, 4, 4-pentafluorobutene; 6-bromo-5, 5,6, 6-tetrafluorohexene; 4-bromoperfluorobutene-1 and 3, 3-difluoroallyl bromide. Brominated vinyl ether cure site monomers include 2-bromo-perfluoroethyl perfluorovinyl ether and CF₂Br-R_f-O-CF＝CF₂(wherein R is_fBeing a perfluoroalkylene group), e.g. CF₂BrCF₂O-CF＝CF₂And ROCF ═ CFBr or ROCBR ═ CF₂(wherein R is a lower alkyl or fluoroalkyl group) fluorovinylethers, e.g. CH₃OCF ═ CFBr or CF₃CH₂OCF＝CFBr。

Suitable iodinated cure site monomers include iodinated olefins of the general formula: CHR ═ CH-Z-CH₂CHR-I, wherein R is-H or-CH₃(ii) a Z is C₁-C₁₈(per) fluoroalkylene groups, linear or branched, optionally containing one or more ether oxygen atoms, or (per) fluoropolyoxyalkylene groups, as disclosed in U.S. Pat. No.5,674,959. Useful iodinated cure site units Other examples of bodies are unsaturated ethers of the general formula: i (CH)₂CF₂CF₂)_nOCF＝CF₂And ICH₂CF₂O[CF(CF₃)CF₂O]_nCF＝CF₂Etc., where n is an integer from 1 to 3, as disclosed in U.S. patent No.5,717,036. Additionally, ethylene iodide, 4-iodo-3, 3, 4, 4-tetrafluorobutene-1 (ITFB); 3-chloro-4-iodo-3, 4, 4-trifluorobutene; 2-iodo-1, 1, 2, 2-tetrafluoro-1- (vinyloxy) ethane; 2-iodo-1- (perfluorovinyloxy) -1, 1, 2, 2-tetrafluoroethylene; 1, 1, 2, 3, 3, 3-hexafluoro-2-iodo-1- (perfluorovinyloxy) propane; 2-iodoethyl vinyl ether; 3, 3, 4, 5, 5, 5-hexafluoro-4-iodopentene; suitable iodinated cure site monomers, and monoiodotrifluoroethylene, are disclosed in U.S. Pat. No.4,694,045. Allyl iodide and 2-iodo-perfluoroethyl perfluorovinyl ether are also useful cure site monomers.

Useful nitrile-containing cure site monomers include those of the general formula shown below:

CF₂＝CF-O(CF₂)_n-CN

wherein n is an integer from 2 to 12. In some embodiments, n is an integer from 2 to 6.

CF₂＝CF-O[CF₂-CF(CF)-O]_n-CF₂-CF(CF₃)-CN

Wherein n is an integer from 0 to 4. In some embodiments, n is an integer from 0 to 2.

CF₂＝CF-[OCF₂CF(CF₃)]_x-O-(CF₂)_n-CN

Wherein x is 1 or 2, and n is an integer from 1 to 4; and

CF₂＝CF-O-(CF₂)_n-O-CF(CF₃)-CN

wherein n is an integer from 2 to 4. In some embodiments, the cure site monomer is a perfluorinated polyether having nitrile groups and trifluorovinyl ether groups.

In some embodiments, the cure site monomer is:

CF₂＝CFOCF₂CF(CF₃)OCF₂CF₂CN

i.e., perfluoro (8-cyano-5-methyl-3, 6-dioxa-1-octene) or 8-CNVE.

Examples of non-conjugated diene cure site monomers include, but are not limited to, 1, 4-pentadiene; 1, 5-hexadiene; 1, 7-octadiene; 3, 3, 4, 4-tetrafluoro-1, 5-hexadiene; and other similar monomers such as those disclosed in canadian patent No.2,067,891 and european patent No. 0784064A1. A suitable triene is 8-methyl-4-ethylene-1, 7-octadiene.

In embodiments where the fluoroelastomer is cured by peroxide, the cure site monomer is preferably selected from 4-bromo-3, 3, 4, 4-tetrafluorobutene-1 (BTFB); 4-iodo-3, 3, 4, 4-tetrafluorobutene-1 (ITFB); allyl iodide; bromotrifluoroethylene and 8-CNVE. In embodiments where the fluoroelastomer is cured with a polyol, 2HPFP or perfluoro (2-phenoxypropyl vinyl) ether are preferred cure site monomers. In embodiments where the fluoroelastomer is cured with tetraamine, bis (aminophenol) or bis (thioaminophenol), 8-CNVE, is the preferred cure site monomer.

The cure site monomer units, when present in the presently disclosed fluoroelastomers, are typically present at a level of from 0.05 to 10 weight percent (based on the total weight of the fluoroelastomer), preferably from 0.05 to 5 weight percent and most preferably from 0.05 to 3 weight percent.

Fluoroelastomers useful in the presently disclosed subject matter include, but are not limited to, those having at least 58 weight percent fluorine and comprising copolymerized units of: i) vinylidene fluoride and hexafluoropropylene; ii) vinylidene fluoride, hexafluoropropylene and tetrafluoroethylene; iii) vinylidene fluoride, hexafluoropropylene, tetrafluoroethylene, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1; iv) vinylidene fluoride, hexafluoropropylene, tetrafluoroethylene, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1; v) vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1; vi) vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1; vii) vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 1, 1, 3, 3, 3-pentafluoropropene; viii) tetrafluoroethylene, perfluoro (methyl vinyl) ether and ethylene; ix) tetrafluoroethylene, perfluoro (methyl vinyl) ether, ethylene and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1; x) tetrafluoroethylene, perfluoro (methyl vinyl) ether, ethylene and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1; xi) tetrafluoroethylene, propylene and vinylidene fluoride; xii) tetrafluoroethylene and perfluoro (methyl vinyl) ether; xiii) tetrafluoroethylene, perfluoro (methyl vinyl) ether and perfluoro (8-cyano-5-methyl-3, 6-dioxa-1-octene); xiv) tetrafluoroethylene, perfluoro (methyl vinyl) ether, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1; xv) tetrafluoroethylene, perfluoro (methyl vinyl) ether and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1; and xvi) tetrafluoroethylene, perfluoro (methyl vinyl) ether and perfluoro (2-phenoxypropyl vinyl) ether.

In addition, due to the use of chain transfer agents or molecular weight regulators during the preparation of the fluoroelastomer, iodine-containing end groups, bromine-containing end groups or combinations thereof can optionally be present at one or both ends of the fluoroelastomer polymer chain ends. When used, the amount of chain transfer agent is calculated so that the level of iodine or bromine in the fluoroelastomer is in the range of 0.005-5 wt.%, preferably 0.05-3 wt.%.

Examples of the chain transfer agent include iodine-containing compounds such that iodine is introduced into the bond at one or both ends of the polymer molecule. Diiodomethane; 1, 4-diiodoperfluoro-n-butane; and 1, 6-diiodo-3, 3, 4, 4-tetrafluorohexane are representative of such agents. Other iodinated chain transfer agents include 1, 3-diiodoperfluoropropane; 1, 6-diiodoperfluorohexane; 1, 3-diiodo-2-chloroperfluoropropane; 1, 2-bis (iododifluoromethyl) perfluorocyclobutane; monoiodoperfluoroethane; monoiodoperfluorobutane; 2-iodo-1-hydroperfluoroethane, and the like. Also included are cyano-iodo chain transfer agents disclosed in European patent No. 0868447A1. Particularly preferred are di-iodinated chain transfer agents.

Examples of brominated chain transfer agents include 1-bromo-2-iodoperfluoroethane; 1-bromo-3-iodoperfluoropropane; 1-iodo-2-bromo-1, 1-difluoroethane and other analogs, as disclosed in US patent No.5,151,492.

Other chain transfer agents suitable for use include those disclosed in U.S. Pat. No.3,707,529. Examples of such agents include isopropanol, diethyl malonate, ethyl acetate, carbon tetrachloride, acetone and dodecyl mercaptan.

Method of forming a microfluidic device by a thermal radical cure process

In some embodiments, the presently disclosed subject matter provides a method of forming a microfluidic device by contacting a functionalized liquid perfluoropolyether (PFPE) precursor material with a patterned substrate, i.e., master, and then thermally curing using a free radical initiator. As provided in detail below, in some embodiments, the liquid PFPE precursor material is fully cured to form a fully cured PFPE network, which is then removed from the patterned substrate and contacted with a second substrate to form a reversible hermetic seal.

In some embodiments, the liquid PFPE precursor material is partially cured to form a partially cured PFPE network. In some embodiments, the partially cured network is in contact with a second partially cured layer of PFPE material, and then the curing reaction proceeds to completion, thereby forming a permanent bond between the two PFPE layers.

In addition, the partially cured PFPE network can be contacted with a layer or substrate comprising another polymeric material, such as poly (dimethylsiloxane) or another polymer, and then thermally cured to adhere the PFPE network to the other polymeric material. In addition, the partially cured PFPE network can be contacted with a solid substrate such as glass, quartz or silicon and bonded to the substrate by using a silane coupling agent.

Method of forming a patterned layer of elastomeric material

In some embodiments, the presently disclosed subject matter provides methods of forming patterned layers of elastomeric materials. The presently disclosed methods are suitable for use with the perfluoropolyether materials described in section ii.a. and ii.b. and the fluoroolefin-based materials described in section ii.c. The advantage of using a higher viscosity PFPE material is that a higher molecular weight between the crosslinking points is possible. Higher molecular weights between the crosslinking points can improve the elastic properties of the material, preventing the formation of cracks. Referring now to fig. 1A-1C, schematic diagrams of embodiments of the presently disclosed subject matter are shown. A substrate 100 having a patterned surface 102 comprising raised protrusions 104 is depicted. Thus, the patterned surface 102 of the substrate 100 includes at least one raised protrusion 104 that forms the shape of the pattern. In some embodiments, patterned surface 102 of substrate 100 includes a plurality of raised protrusions 104 that form a complex pattern.

As best seen in fig. 1B, a liquid precursor material 106 is disposed on the patterned surface 102 of the substrate 100. As shown in fig. 1B, the liquid precursor material 106 is treated with a treatment process Tr. After treatment of the liquid precursor material 106, a patterned layer 108 of elastomeric material is formed (as shown in fig. 1C).

As shown in fig. 1C, patterned layer 108 of elastomeric material includes pockets 110 formed in a bottom surface of patterned layer 108. The dimensions of the recesses 110 correspond to the dimensions of the raised protrusions 104 of the patterned surface 102 of the substrate 100. In some embodiments, pocket 110 includes at least one channel 112, which in some embodiments of the presently disclosed subject matter includes a microscale channel. Patterned layer 108 is removed from patterned surface 102 of substrate 100 to provide microfluidic device 114.

In some embodiments, the patterned substrate comprises an etched silicon wafer. In some embodiments, the patterned substrate comprises a patterned substrate of photoresist. For purposes of the presently disclosed subject matter, the patterned substrate can be fabricated by any of the processing methods known in the art, including, but not limited to, photolithography, electron beam lithography, and ion milling (ion milling).

In some embodiments, the patterned layer of perfluoropolyether has a thickness of between about 0.1 micron and about 100 microns. In some embodiments, the patterned layer of perfluoropolyether has a thickness of between about 0.1 mm and about 10 mm. In some embodiments, the patterned layer of perfluoropolyether has a thickness of between about 1 micron and about 50 microns. In some embodiments, the patterned layer of perfluoropolyether has a thickness of about 20 micrometers. In some embodiments, the patterned layer of perfluoropolyether has a thickness of about 5 mm.

In some embodiments, the patterned layer of perfluoropolyether includes a plurality of micron-scale channels. In some embodiments, the channel has a width from about 0.01 μm to about 1000 μm; a width from about 0.05 μm to about 1000 μm; and/or a width from about 1 μm to about 1000 μm. In some embodiments, the channel has a width from about 1 μm to about 500 μm; a width from about 1 μm to about 250 μm; and/or a width from about 10 μm to about 200 μm. Exemplary channel widths include, but are not limited to, 0.1 μm, 1 μm, 2 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120 μm, 130 μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm, 200 μm, 210 μm, 220 μm, 230 μm, 240 μm, and 250 μm.

In some embodiments, the channel has a depth ranging from about 1 μm to about 1000 μm; and/or a depth range from about 1 μm to 100 μm. In some embodiments, the channel has a depth ranging from about 0.01 μm to about 1000 μm; a depth range from about 0.05 μm to about 500 μm; a depth range from about 0.2 μm to about 250 μm; a depth range from about 1 μm to about 100 μm; a depth range from about 2 μm to about 20 μm; and/or a depth range from about 5 μm to about 10 μm. Exemplary channel depths include, but are not limited to, 0.01 μm, 0.02 μm, 0.05 μm, 0.1 μm, 0.2 μm, 0.5 μm, 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 7.5 μm, 10 μm, 12.5 μm, 15 μm, 17.5 μm, 20 μm, 22.5 μm, 25 μm, 30 μm, 40 μm, 50 μm, 75 μm, 100 μm, 150 μm, 200 μm, and 250 μm.

In some embodiments, the channel has a width to depth ratio of from about 0.1: 1 to about 100: 1. In some embodiments, the channel has a width to depth ratio of from about 1: 1 to about 50: 1. In some embodiments, the channel has a width to depth ratio of from about 2: 1 to about 20: 1. In some embodiments, the channel has a width to depth ratio of from about 3: 1 to about 15: 1. In some embodiments, the channel has a width to depth ratio of about 10: 1.

One of ordinary skill in the art will recognize that the dimensions of the channel of the presently disclosed subject matter are not limited to the ranges of the above-described exemplary properties and can be varied in width and depth to affect the amount of force required to flow material through the channel and/or to manipulate the valve to control the flow of material therein. In addition, as described in more detail below, the larger width channels are designed as fluid reservoirs, reaction chambers, mixing channels, separation zones, and the like.

Method of forming multilayer patterned material

In some embodiments, the presently disclosed subject matter describes methods of forming a multilayer patterned material, e.g., a multilayer patterned PFPE material. In some embodiments, multiple layers of patterned perfluoropolyether materials are used to fabricate a monolithic, monolithic PFPE-based microfluidic device.

Referring now to fig. 2A-2D, schematic diagrams of methods of making embodiments of the presently disclosed subject matter are shown. Patterned layers 200 and 202 are provided, each of which, in some embodiments, includes a perfluoropolyether material prepared from a liquid PFPE precursor material having a viscosity greater than about 100 cSt. In this example, each of the patterned layers 200 and 202 includes a plurality of channels 204. In this embodiment of the presently disclosed subject matter, the plurality of channels 204 comprises micron-scale channels. In the patterned layer 200, the channels are indicated by dashed lines, i.e., hatching, in FIGS. 2A-2C. The patterned layer 202 is superimposed on the patterned layer 200 in a predetermined orientation arrangement. In this example, the predetermined orientation is arranged such that the channels in patterned layers 200 and 202 are substantially perpendicular to each other. In some embodiments, as depicted in fig. 2A-2D, patterned layer 200 overlaps unpatterned layer 206. The unpatterned layer 206 may comprise perfluoropolyether.

With continued reference to fig. 2A-2D, patterned layers 200 and 202, and in some embodiments unpatterned layer 206, are processed by a processing method Tr. As described in more detail below, the layers 200, 202, and in some embodiments the unpatterned layer 206, are treated by a treatment process Tr to promote adhesion of the patterned layers 200 and 202 to each other, and in some embodiments, to promote adhesion of the patterned layer 200 to the unpatterned layer 206, as shown in fig. 2C and 2D. The resulting microfluidic device 208 includes an integrated network 210 of microscale channels 204, with the microscale channels 204 intersecting at predetermined intersection points 212, as best seen in the cross-section provided in fig. 2D. Also shown in fig. 2D is a membrane 214 that includes the top surface of the channels 204 of the patterned layer 200, separating the channels 204 of the patterned layer 202 from the channels 204 of the patterned layer 200.

With continued reference to fig. 2A-2C, in some embodiments, patterning layer 202 includes a plurality of apertures, and the apertures are designated as input aperture 216 and output aperture 218. In some embodiments, the apertures, e.g., the input aperture 216 and the output aperture 218, are in fluid communication with the channel 204. In some embodiments, the aperture comprises a side-actuated valve structure comprising a membrane of PFPE material that can be actuated to restrict flow in an adjoining channel (not shown).

In some embodiments, the first patterned layer of photocurable PFPE material is cast at a thickness that imparts a degree of mechanical stability to the PFPE structure. Thus, in some embodiments, the first patterned layer of photocurable PFPE material has a thickness of about 50 μm to several centimeters. In some embodiments, the first patterned layer of photocurable PFPE material has a thickness of between 50 μm and about 10 millimeters. In some embodiments, the first patterned layer of photocurable PFPE material is 5mm thick. In some embodiments, the first patterned layer of photocurable PFPE material is about 4mm thick. Further, in some embodiments, the thickness of the first patterned layer of PFPE material is about 0.1 μm to about 10 cm; from about 1 μm to about 5 cm; from about 10 μm to about 2 cm; and a range from about 100 μm to about 10 mm.

In some embodiments, the second patterned layer of photocurable PFPE material has a thickness of between about 1 micron and about 100 microns. In some embodiments, the second patterned layer of photocurable PFPE material has a thickness of between about 1 micron and about 50 microns. In some embodiments, the second patterned layer of photocurable material has a thickness of about 20 microns.

Although fig. 2A-2C disclose the formation of a microfluidic device in which two patterned layers of PFPE material are combined, it is possible in some embodiments of the presently disclosed subject matter to form a microfluidic device that includes one imaged layer and one unpatterned layer of PFPE material. Thus, the first image layer can include micron-scale channels or an integrated network of micron-scale channels and then the first image layer can be overlaid over the unpatterned layer and bonded to the unpatterned layer using a photocuring step, such as the application of ultraviolet light as disclosed herein, to form a monolithic structure including enclosed channels therein.

Thus, in some embodiments, the first and second patterned layers of photocurable perfluoropolyether material, or alternatively, the first patterned layer of photocurable perfluoropolyether material and the unpatterned layer of photocurable perfluoropolyether material, are bonded to each other, thereby forming an integral monolithic PFPE-based microfluidic device.

Method of forming a patterned PFPE layer by thermal radical cure

In some embodiments, a thermal free radical initiator (including, but not limited to, peroxides and/or azo compounds) is mixed with a liquid perfluoropolyether (PFPE) precursor functionalized with polymerizable groups (including, but not limited to, acrylate, methacrylate, and styrene units) to form a mixture. The mixture is then contacted with a patterned substrate, the "master", and then heated to cure the PFPE precursor into a network, as shown in FIGS. 1A-1C.

In some embodiments, the PFPE precursor is fully cured to form a fully cured PFPE precursor. In some embodiments, the free radical curing reaction proceeds only partially, forming a partially cured network.

Method for bonding a layer of PFPE material to a substrate by a thermal radical curing process

In some embodiments, the fully cured PFPE precursor is peeled (e.g., peeled) from the patterned substrate, i.e., master, and then contacted with a second substrate to form a reversible hermetic seal.

In some embodiments, the partially cured network is in contact with a second partially cured layer of PFPE material, and then the curing reaction proceeds to completion, thereby forming a permanent bond between the two PFPE layers.

In some embodiments, the partial free radical cure process is used to bond at least one layer of partially cured PFPE material to a substrate. In some embodiments, the partial free radical cure process is used to bond multiple layers of partially cured PFPE material to a substrate. In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the substrate is treated with a silane coupling agent.

An embodiment of the presently disclosed method for bonding a layer of PFPE material to a substrate is shown in fig. 3A-3C. Referring now to fig. 3A, a substrate 300 is provided, wherein, in some embodiments, the substrate 300 is selected from the group consisting of a glass material, a quartz material,silicon materials, fused silica materials and plastics. Substrate 300 is passed through treatment process T_r1To be processed. In some embodiments, process T_r1Including treating the substrate with a base/alcohol mixture, such as KOH/isopropanol, to impart hydroxyl functionality to the substrate 300.

Referring now to FIG. 3B, functionalized substrate 300 is coupled with a silane coupling agent, such as R-SiCl₃OR R-Si (OR)₁)₃Wherein R and R₁Indicating that the functional groups described above for forming the silylated substrate 300 are reacting. In some embodiments, the silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein the monohalosilane, dihalosilane, trihalosilane, monoalkoxysilane, dialkoxysilane, and trialkoxysilane are functionalized with a moiety selected from the group consisting of amines, methacrylates, acrylates, styrenes, epoxy, isocyanate, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

Referring now to FIG. 3C, silanized substrate 300 is contacted with a patterned layer of partially cured PFPE material 302 and treated by treatment T_r2Treated to form a permanent bond between the patterned layer of PFPE material 302 and the substrate 300.

In some embodiments, partial free radical cure is used to bond the PFPE layer to a second polymeric material such as poly (dimethylsiloxane) (PDMS) materials, polyurethane materials, polysiloxane-containing polyurethane materials, and PFPE-PDMS block copolymer materials. In some embodiments, the second polymeric material comprises a functionalized polymeric material. In some embodiments, the second polymeric material is terminated with a polymerizable group. In some embodiments, the polymerizable group is selected from acrylates, styrenes, and methacrylates. Further, in some embodiments, the second polymeric material is treated with a plasma and a silane coupling agent to introduce desired functional groups into the second polymeric material.

An embodiment of the presently disclosed method for bonding a patterned layer of PFPE material to another patterned layer of polymeric material is illustrated in fig. 4A-4C. Referring now to fig. 4A, a patterned layer of a first polymeric material 400 is provided. In some embodiments, the first polymeric material comprises a PFPE material. In some embodiments, the first polymeric material comprises a polymeric material selected from the group consisting of a poly (dimethylsiloxane) material, a polyurethane material, a polysiloxane-containing polyurethane material, and a PFPE-PDMS block copolymer material. The patterned layer of the first polymer material 400 is processed through the process Tr 1. In some embodiments, process T_r1Is comprised in O₃And exposing the patterned layer of the first polymeric material 400 to UV light in the presence of the R functional group to add the R functional group to the patterned layer of the polymeric material 400.

Referring now to FIG. 4B, the functionalized patterned layer of first polymeric material 400 is contacted with the top surface of the functionalized patterned layer of PFPE material 402, and then passed through treatment process T_r2Processed to form a two-layer hybrid assembly 404. Thus, the functionalized patterned layer of first polymeric material 400 is bonded to the functionalized patterned layer of PFPE material 402.

Referring now to fig. 4C, in some embodiments, the two-layer hybrid component 404 is contacted with a substrate 406 to form a multilayer hybrid structure 410. In some embodiments, the substrate 406 is coated with a coating of liquid PFPE precursor material 408. The multilayer hybrid structure 410 is passed through a process T_r3To bond the two-layer assembly 404 to the substrate 406.

Method of forming a microfluidic device by a two-component curing process

The presently disclosed subject matter provides methods of forming microfluidic devices by which a functionalized perfluoropolyether (PFPE) precursor is contacted with a patterned surface and then cured by reaction of two components such as epoxy/amine, hydroxyl/isocyanate, hydroxyl/acid chloride, and hydroxyl/chlorosilane to form a fully cured or partially cured PFPE network. In some embodiments, the partially cured PFPE network is contacted with another substrate and then curing is allowed to proceed to completion in order to bond the PFPE network to the substrate. This method can be used to bond multiple layers of PFPE material to a substrate.

Further, in some embodiments, the substrate comprises a second polymeric material, such as PDMS, or another polymer. In some embodiments, the second polymeric material comprises an elastomer other than PDMS, such as Kraton, buna rubber, natural rubber, fluoroelastomer, chloroprene, butyl rubber, nitrile rubber, polyurethane, or a thermoplastic elastomer. In some embodiments, the second polymeric material comprises a rigid thermoplastic material, including but not limited to: polystyrene, poly (methyl methacrylate), polyesters, such as poly (ethylene terephthalate), polycarbonates, polyimides, polyamides, polyvinyl chloride, polyolefins, poly (ketones), poly (ether ketones), and poly (ether sulfones).

In some embodiments, the PFPE layer is adhered to a solid substrate, such as a glass material, a quartz material, a silicon material, and a fused silica material, using a silane coupling agent.

Method for forming patterned PFPE layer by two-component curing method

In some embodiments, the PFPE network is formed by the reaction of a two-component functionalized liquid precursor system. Utilizing the general method of forming a patterned layer of polymeric material as shown in fig. 1A-1C, a liquid precursor material comprising a two-component system is contacted with a patterned substrate and a patterned layer of PFPE material is formed. In some embodiments, the two-part liquid precursor system is selected from the group consisting of epoxy/amine systems, hydroxyl/isocyanate systems, amine/isocyanate systems, hydroxyl/acid chloride systems, and hydroxyl/chlorosilane systems. The functionalized liquid precursor is mixed in a suitable ratio and then contacted with the patterned surface or master. The curing reaction is carried out by using heat, a catalyst, etc. until a network is formed.

In some embodiments, a fully cured PFPE precursor is formed. In some embodiments, the two-component reaction proceeds only partially, thereby forming a partially cured PFPE network.

Method for bonding PFPE layer to substrate by two-component curing process

IV.B.1. full cure with two-component curing Process

In some embodiments, the fully cured PFPE two-component precursor is removed (e.g., peeled off) from the master and then contacted with a substrate to form a reversible hermetic seal. In some embodiments, the partially cured network is contacted with another partially cured layer of the PFPE, and then the reaction proceeds to completion, thereby forming a permanent bond between the two layers.

IV.B.2. partial curing of two-component systems

As shown in fig. 3A-3C, in some embodiments, the two-part partial cure process is used to bond at least one layer of a partially cured PFPE material to a substrate. In some embodiments, the two-part partial cure process is used to bond multiple layers of partially cured PFPE material to a substrate. In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the substrate is treated with a silane coupling agent.

As in fig. 4A-4C, in some embodiments, a two-part partial cure is used to bond the PFPE layer to a second polymeric material, such as a poly (dimethylsiloxane) (PDMS) material. In some embodiments, the PDMS material comprises a functionalized PDMS material. In some embodiments, PDMS is treated with a plasma and a silane coupling agent to introduce desired functional groups into the PDMS material. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable group comprises an epoxy group. In some embodiments, the polymerizable group comprises an amine.

In some embodiments, the second polymeric material comprises an elastomer other than PDMS, such as Kraton, buna rubber, natural rubber, fluoroelastomer, chloroprene, butyl rubber, nitrile rubber, polyurethane, or a thermoplastic elastomer. In some embodiments, the second polymeric material comprises a rigid thermoplastic material, including but not limited to: polystyrene, poly (methyl methacrylate), polyesters, such as poly (ethylene terephthalate), polycarbonates, polyimides, polyamides, polyvinyl chloride, polyolefins, poly (ketones), poly (ether ketones), and poly (ether sulfones).

IV.B.3. overcuring of two-component systems

The presently disclosed subject matter provides methods of forming microfluidic devices by which a functionalized perfluoropolyether (PFPE) precursor is contacted with a patterned substrate and then cured by reaction of two components such as epoxy/amine, hydroxyl/isocyanate, hydroxyl/acid chloride, and hydroxyl/chlorosilane to form a layer of cured PFPE material. In this particular method, the layer of cured PFPE material is capable of bonding to a second substrate by fully curing the layer with an excess of one component and contacting the layer of cured PFPE material with a second substrate comprising an excess of a second component to react the excess of these groups to bond the two layers.

Thus, in some embodiments, two-component systems are mixed, such as epoxy/amine systems, hydroxyl/isocyanate systems, amine/isocyanate systems, hydroxyl/acid chloride systems, or hydroxyl/chlorosilane systems. In some embodiments, at least one component of the two-component system is in excess relative to the other component. The reaction is then carried to completion by heating, use of a catalyst, or the like, with the remaining cured network including many functional groups resulting from the presence of excess components.

In some embodiments, two layers of a fully cured PFPE material comprising complementary excess groups are contacted with each other, wherein the excess groups react, thereby forming a permanent bond between the two layers.

As shown in fig. 3A-3C, in some embodiments, a fully cured PFPE network comprising an excess of functional groups is contacted with a substrate. In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the substrate is treated with a silane coupling agent such that the functional groups on the coupling agent are complementary to the excess functional groups on the fully cured network. Thus, a permanent bond is formed on the substrate.

As shown in fig. 4A-4C, in some embodiments, a two-part overcure is used to bond the PFPE network to a second polymeric material, such as a poly (dimethylsiloxane) PDMS material. In some embodiments, the PDMS material comprises a functionalized PDMS material. In some embodiments, the PDMS material is treated with a plasma and a silane coupling agent to introduce the desired functional groups. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable material comprises an epoxy group. In some embodiments, the polymerizable material comprises an amine.

Method for functionalizing surfaces of micro-and/or nano-scale devices

In some embodiments, the presently disclosed subject matter provides materials and methods for functionalizing channels in microfluidic devices and/or microtitre tubes. In some embodiments, such functionalization includes, but is not limited to, synthesis of peptides and other natural polymers and/or binding to the inner surface of channels in microfluidic devices. Thus, the presently disclosed subject matter can be used in microfluidic devices such as those described by Rolland, J.et al, JACS 2004, 126, 2322-2323, the disclosure of which is incorporated herein by reference in its entirety.

In some embodiments, the method comprises binding the small molecule to an inner surface of a microfluidic channel or a surface of a microtiter tube. In such embodiments, the small molecule, once bound, can serve as a variety of functional groups. In some embodiments, the small molecule serves as a cleavable group that, upon activation, is capable of changing the polarity of the channel and thus increasing the wettability of the channel. In some embodiments, the small molecule serves as a binding site. In some embodiments, the small molecule serves as a binding site for one of a catalyst, a drug, a substrate for a drug, an analyte, and a sensor. In some embodiments, the small molecule serves as a reactive functional group. In some embodiments, the reactive functional group reacts to give a zwitterion. In some embodiments, the zwitterion provides a polar, ionic pathway.

An embodiment of the presently disclosed method for functionalizing the interior surface of a microfluidic channel and/or a microtiter tube is illustrated in fig. 5A and 5B. Referring now to fig. 5A, a microfluidic channel 500 is provided. In some embodiments, microfluidic channel 500 is formed from a functionalized PFPE material comprising R functional groups as described herein. In some embodiments, microchannel 500 comprises a PFPE network that undergoes a post-cure treatment process in which functional groups R are introduced into the interior surface 502 of microfluidic channel 500.

Referring now to fig. 5B, a micro titer tube 504 is provided. In some embodiments, the microtiter tube 504 is coated with a layer of functionalized PFPE material 506 that includes R functional groups to impart functional groups to the microtiter tube 504.

Method for connecting V.A. functional group to PFPE network

In some embodiments, a PFPE network comprising excess functional groups is used to functionalize the inner surface of a microfluidic channel or the surface of a microtiter tube. In some embodiments, the interior surface of the microfluidic channel or the surface of the microtiter tube is functionalized by attaching a functionalizing moiety selected from the group consisting of proteins, oligonucleotides, drugs, ligands, catalysts, dyes, sensors, analytes, and charged species capable of altering the wettability of the channel.

In some embodiments, a latent functional group is introduced into the fully cured PFPE network. In some embodiments, latent methacrylate groups are present on the surface of the PFPE network that has been free-radically cured photochemically or thermally. The multiple layers of fully cured PFPE are then contacted with the functionalized surface of the PFPE network, forming a seal and reacted by, for example, heating to allow the latent functional groups to react and form a permanent bond between the layers.

In some embodiments, the latent functional groups photochemically react with each other at wavelengths different from those used to cure the PFPE precursors. In some embodiments, this method is used to bond a fully cured layer to a substrate. In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the substrate is treated with a silane coupling agent that is complementary to the latent functional group.

In some embodiments, such latent functional groups serve to bond the fully cured PFPE network to a second polymeric material, such as a poly (dimethylsiloxane) PDMS material. In some embodiments, the PDMS material comprises a functionalized PDMS material. In some embodiments, PDMS is treated with a plasma and a silane coupling agent to introduce desired functional groups. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable group is selected from acrylates, styrenes, and methacrylates.

Method for introducing functional groups in the production of liquid PFPE precursors

The presently disclosed subject matter provides methods of forming a microfluidic device by which a photochemically cured PFPE layer is brought into contact with a second substrate, thereby forming a seal. The PFPE layer is then heated at elevated temperatures to bond the layer to the substrate via the latent functional groups. In some embodiments, the second substrate further comprises a cured PFPE layer. In some embodiments, the second substrate comprises a second polymeric material, such as a poly (dimethylsiloxane) (PDMS) material.

In some embodiments, the latent group comprises a methacrylate unit, which is not reactive during photocuring. Furthermore, in some embodiments, the latent group is introduced in the production of the liquid PFPE precursor. For example, in some embodiments, methacrylate units are added to PFPE diols through the use of glycidyl methacrylate, and the reaction of hydroxyl and epoxy groups produces secondary alcohols, which can be used as a means of introducing chemical functionality. In some embodiments, multiple layers of fully cured PFPE are bonded to each other through these latent functional groups. In some embodiments, the latent functional groups serve to bond the fully cured PFPE layer to the substrate. In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the substrate is treated with a silane coupling agent.

In addition, this method can be used to bond a fully cured PFPE layer to a second polymeric material, such as a poly (dimethylsiloxane) (PDMS) material. In some embodiments, the PDMS material comprises a functionalized PDMS material. In some embodiments, PDMS is treated with a plasma and a silane coupling agent to introduce desired functional groups. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable material is selected from the group consisting of acrylates, styrenes, and methacrylates.

In some embodiments, a PFPE network containing latent functional groups is used to functionalize the inner surface of a microfluidic channel or microtiter tube. Examples include the attachment of proteins, oligonucleotides, drugs, ligands, catalysts, dyes, sensors, analytes, and charged species capable of altering the wettability of a channel.

Method for linking multiple chains of PFPE material with functionalized linker groups

In some embodiments, the presently disclosed methods add functional groups to a microfluidic channel or microtitre tube by adding chemical "linker" moieties to the elastomer itself. In some embodiments, functional groups are added along the backbone of the precursor material. An example of this method is shown in flow 6.

Scheme 6. representative methods for adding functional groups along the backbone of the precursor Material

In some embodiments, the precursor material comprises a macromolecule comprising hydroxyl functional groups. In some embodiments, as depicted in scheme 6, the hydroxyl functional groups comprise diol functional groups. In some embodiments, two or more of the diol functional groups are linked by a trifunctional "linker" molecule. In some embodiments, the trifunctional linker molecule has two functional groups, R and R'. In some embodiments, the R' group is reactive with a hydroxyl group of the macromolecule. In scheme 6, the ring may represent a linker molecule; and the wavy line may represent a PFPE chain.

In some embodiments, the R group provides a desired functional group to the inner surface of a microfluidic channel or the surface of a microtiter tube. In some embodiments, the R' group is selected from the following non-limiting groups: acid chlorides, isocyanates, halogens and ester moieties. In some embodiments, the R group is selected from one of a protected amine and a protected alcohol, but is not limited thereto. In some embodiments, the macrodiol is functionalized with polymerizable methacrylate groups. In some embodiments, the functionalized macromolecule is cured and/or molded by a photochemical process described by Rolland, J et al, JACS 2004, 126, 2322-2323, the disclosure of which is incorporated herein by reference in its entirety.

Thus, the presently disclosed subject matter provides a method of adding latent functional groups to a photocurable PFPE material via functionalized linker groups. Thus, in some embodiments, multiple chains of PFPE material are linked together before the chains are terminated by a polymerizable group. In some embodiments, the polymerizable group is selected from the group consisting of methacrylates, acrylates, and styrenes. In some embodiments, the latent functional groups are chemically attached to the "linker" molecules such that they are present in the fully cured network.

In some embodiments, the latent functional groups introduced in this manner are used to bond multiple layers of PFPE, to bond a fully cured PFPE layer to a substrate, such as a glass or silicon material that has been treated with a silane coupling agent, or to bond a fully cured PFPE layer to a second polymeric material, such as a PDMS material. In some embodiments, the PDMS material is treated with a plasma and a silane coupling agent to introduce the desired functional groups. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable group is selected from acrylates, styrenes, and methacrylates.

In some embodiments, a PFPE network comprising functional groups attached to "linker" molecules may be used to functionalize the inner surface of a microfluidic channel and/or the surface of a microtiter tube. In some embodiments, the interior of the microfluidic channel is functionalized by attaching a functionalization moiety selected from the group consisting of a protein, an oligonucleotide, a drug, a ligand, a catalyst, a dye, a sensor, an analyte, and a charged species capable of altering the wettability of the channel.

Method for adding functionalized monomers to PFPE precursor materials

In some embodiments, the method includes adding a functionalized monomer to the uncured precursor material. In some embodiments, the functionalized monomer is selected from functionalized styrenes, methacrylates, and acrylates. In some embodiments, the precursor material comprises a fluoropolymer. In some embodiments, the functionalized monomer comprises a highly fluorinated monomer. In some embodiments, the highly fluorinated monomer comprises perfluoroethyl vinyl ether (EVE). In some embodiments, the precursor material comprises a poly (dimethylsiloxane) (PDMS) elastomer. In some embodiments, the precursor material comprises a polyurethane elastomer. In some embodiments, the method further comprises adding the functionalized monomer to the network through a curing step.

In some embodiments, the functional monomer is added directly to the liquid PFPE precursor to be incorporated into the network after crosslinking. For example, monomers may be introduced into the network capable of reaction-post crosslinking in order to bond multiple layers of PFPE to, bond a fully cured PFPE layer to a substrate such as a glass or silicon material that has been treated with a silane coupling agent, or bond a fully cured PFPE layer to a second polymeric material such as a PDMS material. In some embodiments, the PDMS material is treated with a plasma and a silane coupling agent to introduce the desired functional groups. In some embodiments, the PDMS material is end-capped with a polymerizable group. In some embodiments, the polymerizable material is selected from the group consisting of acrylates, styrenes, and methacrylates.

In some embodiments, a functionalized monomer is added directly to the liquid PFPE precursor and used to attach a functionalized moiety selected from the group consisting of proteins, oligonucleotides, drugs, ligands, catalysts, dyes, sensors, analytes, and charged species capable of altering the wettability of the channel.

Such monomers include, but are not limited to, t-butyl methacrylate, t-butyl acrylate, dimethylaminopropyl methacrylate, glycidyl methacrylate, hydroxyethyl methacrylate, aminopropyl methacrylate, allyl acrylate, cyanoacrylate, cyanomethacrylate, trimethoxysilane acrylate, trimethoxysilane methacrylate, isocyanatomethacrylate, lactone-containing acrylates and methacrylates, sugar-containing acrylates and methacrylates, polyethylene glycol methacrylate, norbornane-containing methacrylates and acrylates, polyhedral oligomeric silsesquioxane methacrylate, 2-trimethylsiloxyethyl methacrylate, 1H, 1H, 2H, 2H-fluorooctyl methacrylate, pentafluorostyrene, vinylpyridine, bromostyrene, chlorostyrene, styrenesulfonic acid, fluorostyrene, styrene acetate, acrylamide, and acrylonitrile.

In some embodiments, the monomer to which the above reagents have been attached is mixed directly with the liquid PFPE precursor to be introduced into the network after crosslinking. In some embodiments, the monomer includes a group selected from a polymerizable group, a desired agent, and a fluorinated moiety to provide miscibility with the PFPE liquid precursor. In some embodiments, the monomer does not include polymerizable groups, desired agents, and fluorinated moieties required for miscibility with the PFPE liquid precursor.

In some embodiments, monomers are added to adjust the mechanical properties of the fully cured elastomer. Such monomers include, but are not limited to: perfluoro (2, 2-dimethyl-1, 3-dioxole), hydrogen bonding monomers containing hydroxyl, urethane, urea or other such moieties, monomers containing bulky side groups such as t-butyl methacrylate.

In some embodiments, functionalized species such as the monomers described above are incorporated and mechanically entangled, i.e., non-covalently bonded into the network, after curing. For example, in some embodiments, functional groups are introduced into the PFPE chains that do not contain polymerizable monomers and such monomers are mixed with the curable PFPE species. In some embodiments, such entanglement materials may be used to bond multiple layers of the cured PFPE together if two materials are reactive, such as: epoxy/amine, hydroxyl/acid chloride, hydroxyl/isocyanate, amine/halide, hydroxyl/halide, amine/ester, and amine/carboxylic acid. Upon heating, the functional groups will react and bond the two layers together.

Additionally, such entangled substances may be used to bond a PFPE layer to a layer of another material, such as glass, silicon, quartz, PDMS, Kratons, buna rubber, natural rubber, fluoroelastomers, chloroprene, butyl rubber, nitrile rubber, polyurethane, or thermoplastic elastomers. In some embodiments, the second polymeric material comprises a rigid thermoplastic material, including but not limited to: polystyrene, poly (methyl methacrylate), polyesters, such as poly (ethylene terephthalate), polycarbonates, polyimides, polyamides, polyvinyl chloride, polyolefins, poly (ketones), poly (ether ketones), and poly (ether sulfones).

In some embodiments, such entangled materials may be used to functionalize the interior of a microfluidic channel for the purposes described above.

Other methods of introducing functional groups to the surface of PFPE

In some embodiments, the argon plasma isIs used to make by usingChen, Y. andMomose，Y.interfacial, anal.1999, 27, 1073-. More specifically, without being bound to any one particular theory, fully cured PFPE materials may incorporate functional groups along the fluorinated backbone after exposure to an argon plasma for a period of time.

Such functional groups may be used to bond multiple layers of PFPE, to bond a fully cured PFPE layer to a substrate, such as a glass or silicon material that has been treated with a silane coupling agent, or to bond a fully cured PFPE layer to a second polymeric material, such as a PDMS material. In some embodiments, the PDMS material comprises a functionalized material. In some embodiments, the PDMS material is treated with a plasma and a silane coupling agent to introduce the desired functional groups. Such functional groups may also be used to link proteins, oligonucleotides, drugs, catalysts, dyes, sensors, analytes and charged species capable of altering the wettability of a channel.

In some embodiments, the fully cured PFPE layer is conformed in contact with a solid substrate. In some embodiments, the solid substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. In some embodiments, the PFPE material is irradiated with UV light, such as 185-nm UV light, which is capable of abstracting fluorine atoms from the backbone and forming chemical bonds with the substrate, as described in Vurens, G. et al Langmuir 1992, 8, 1165-. Thus, in some embodiments, the PFPE layer is covalently bonded to the solid substrate by radical coupling following the abstraction of the fluorine atom.

Bonding of micro-or nano-scale devices to substrates by means of encapsulating polymers

In some embodiments, micro-scale devices, nano-scale devices, or a combination thereof are bonded to a substrate by placing a fully cured device in conforming contact with the substrate and pouring an "encapsulating polymer" over the entire device. In some embodiments, the encapsulating polymer is selected from the group consisting of liquid epoxy precursors and polyurethanes. The encapsulating polymer is then solidified by curing or other means. The encapsulation serves to mechanically bond the layers together and to bond the layers to the substrate.

In some embodiments, the microdevice, the nanodevice, or a combination thereof includes one of the perfluoropolyether materials described in section ii.a and section ii.b above and the fluoroolefin-based materials described in section ii.c above.

In some embodiments, the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused quartz material, and a plastic. Further, in some embodiments, the substrate comprises a second polymeric material, such as poly (dimethylsiloxane) (PDMS), or another polymer. In some embodiments, the second polymeric material comprises an elastomer other than PDMS, such as Kraton, buna rubber, natural rubber, fluoroelastomer, chloroprene, butyl rubber, nitrile rubber, polyurethane, or a thermoplastic elastomer. In some embodiments, the second polymeric material comprises a rigid thermoplastic material, including but not limited to: polystyrene, poly (methyl methacrylate), polyesters, such as poly (ethylene terephthalate), polycarbonates, polyimides, polyamides, polyvinyl chloride, polyolefins, poly (ketones), poly (ether ketones), and poly (ether sulfones). In some embodiments, the surface of the substrate is functionalized with a silane coupling agent such that it reacts with the encapsulating polymer to form an irreversible bond.

IX. method for forming micron-scale structures using sacrificial layers

The presently disclosed subject matter provides methods of forming microchannels or microscale structures for use as microfluidic devices by using sacrificial layers comprising degradable or selectively soluble materials. In some embodiments, the method includes contacting a liquid precursor material with a two-dimensional or three-dimensional sacrificial structure, treating (e.g., curing) the precursor material, and removing the sacrificial structure to form a microfluidic channel.

Thus, in some embodiments, the PFPE liquid precursor is disposed on a multi-dimensional scaffold (multi-dimensional scaffold), wherein the multi-dimensional scaffold is fabricated from a material that is capable of being degraded or washed away after curing of the PFPE network. These materials protect the channels from being filled when another elastomeric layer is cast over them. Examples of such degradable or selectively soluble materials include, but are not limited to, waxes, photoresists, polysulfones, polylactones, cellulosic fibers, salts, or any solid organic or inorganic compound. In some embodiments, the sacrificial layer is removed thermally, photochemically, or by washing with a solvent. Importantly, the compatibility of the materials and devices disclosed herein with organic solvents provides the ability to use sacrificial polymer structures in microfluidic devices.

Such PFPE materials for forming micron-scale structures by using sacrificial layers include those PFPEs and fluoroolefin-based materials described above in section II of the presently disclosed subject matter.

Fig. 6A-6D and 7A-7C illustrate embodiments of the presently disclosed method of forming micron-scale structures by using sacrificial layers of degradable and/or selectively soluble materials.

Referring now to fig. 6A, a patterned substrate 600 is provided. A liquid PFPE precursor material 602 is disposed on the patterned substrate 600. In some embodiments, liquid PFPE precursor material 602 is coated on using spin coatingOn the patterned substrate 600. Liquid PFPE precursor material 602 is passed through treatment process T_r1Processing is performed to form a layer of processed liquid PFPE precursor material 604.

Referring now to fig. 6B, the layer of treated liquid PFPE precursor material 604 is removed from the patterned substrate 600. In some embodiments, the layer of treated liquid PFPE precursor material 604 is in contact with the substrate 606. In some embodiments, the substrate 606 comprises a planar substrate or a substantially planar substrate. In some embodiments, the layer of treated liquid PFPE precursor material is passed through treatment process T_r2Processed to form a two-layer assembly 608.

Referring now to fig. 6C, a predetermined volume of degradable or selectively soluble material 610 is disposed on the two-layer assembly 608. In some embodiments, a predetermined volume of degradable or selectively soluble material 610 is coated on the two-layer assembly 608 using a spin coating process. Referring again to fig. 6C, the liquid precursor material 602 is disposed on a two-layer assembly 608 and processed to form a layer of PFPE material 612 that covers a predetermined volume of degradable or selectively soluble material 610.

Referring now to fig. 6D, a predetermined volume of degradable or selectively soluble material 610 is passed through a treatment process T_r3Treated to remove a predetermined volume of degradable or selectively soluble material 610, thereby forming a micro-scale structure 616. In some embodiments, the micro-scale structures 616 include microfluidic channels. In some embodiments, process T_r3Selected from the group consisting of a heat treatment process, a radiation process and a dissolution process.

In some embodiments, patterned substrate 600 comprises an etched silicon wafer. In some embodiments, the patterned substrate comprises a patterned substrate of photoresist. For purposes of the presently disclosed subject matter, the patterned substrate can be fabricated by any of the processing methods known in the art, including, but not limited to, photolithography, electron beam lithography, and ion milling.

In some embodiments, the degradable or selectively soluble material 610 is selected from the group consisting of polyolefin sulfones, cellulosic fibers, polylactones, and polyelectrolytes. In some embodiments, the degradable or selectively soluble material 610 is selected from materials that are capable of being degraded or dissolved away. In some embodiments, the degradable or selectively soluble material 610 is selected from the group consisting of salts, water soluble polymers and solvent soluble polymers.

In addition to simple channels, the presently disclosed subject matter also provides for the fabrication of multiple complex structures that can be "injection molded" or fabricated in advance and then embedded into the material and removed as described above.

Fig. 7A-C illustrate an embodiment of the presently disclosed method of forming microchannels or microscale structures by using sacrificial layers. Referring now to fig. 7A, a substrate 700 is provided. In some embodiments, substrate 700 is coated with liquid PFPE precursor material 702. Sacrificial structures 704 are located on substrate 700. In some embodiments, liquid PFPE precursor material 702 is passed through treatment process T_r1And (6) processing.

Referring now to fig. 7B, a second liquid PFPE precursor material 706 is disposed on sacrificial structure 704 to encapsulate sacrificial structure 704 in second liquid precursor material 706. The second liquid precursor material 706 is passed through a process T_r2And (6) processing. Referring now to FIG. 7C, sacrificial structure 704 is processed by process T_r3To degrade and/or remove the sacrificial structures, thereby forming the micro-scale structures 708. In some embodiments, the micro-scale structures 708 comprise microfluidic channels.

In some embodiments, the substrate 700 comprises a silicon wafer. In some embodiments, sacrificial structure 704 comprises a degradable or selectively soluble material. In some embodiments, sacrificial structure 704 is selected from the group consisting of a polyalkylene sulfone, a cellulose fiber, a polylactone, and a polyelectrolyte. In some embodiments, the sacrificial structures 704 are selected from materials that can degrade or dissolve away. In some embodiments, sacrificial structure 704 is selected from a salt, a water soluble polymer, and a solvent soluble polymer.

X. microfluidic cell operation

Microfluidic control devices are needed for the development of efficient lab-on-a-chip operations. Valve structures and actuation, fluid control, mixing, separation and detection at the micron level must be designed to transition from large to small. In order to construct such a device, integration of the various components on a common platform must be developed to enable complete control of the solvent and solute.

Microfluidic flow controllers have traditionally been of the external pump type, including hydrodynamic, reciprocating, sonic, and peristaltic types of pumps, and may be simple syringes (see U.S. patent No.6,444,106 to mcbrite et al, U.S. patent No.6,811,385 to Blakley, U.S. published patent application No.20040028566 to Ko et al). Recently, electroosmosis, a method that does not require moving parts, has been successful as a fluid flow driver (see U.S. patent No.6,406,605 to Moles, U.S. patent No.6,568,910 to Parse). Other fluid flow devices that do not require moving parts utilize gravity (see U.S. patent No.6,743,399, Weigl et al), centrifugal force (see U.S. patent No.6,632,388, Sanae et al), capillary action (see U.S. patent No.6,591,852, McNeely et al), or heat (see U.S. published patent application No.20040257668, Ito) to drive the liquid flow through the microchannel. Other inventions generate liquid flow by the application of external forces, such as paddles (see U.S. patent No.6,068,751, issued to Neukermans).

Valves are also used for fluid flow control. The valve can be actuated by applying an external force (e.g., paddle, cantilever, or plug) to the elastomeric channel (see U.S. patent No.6,068,751 to Neukermans). The elastic channels can also contain membranes that can be deflected by pneumatic and/or hydraulic pressure, e.g., hydraulic, electrostatic, or magnetic (see U.S. Pat. No.6,408,878 to Unger et al). Other two-way valves are actuated by light (see U.S. published patent application No.20030156991, Halas et al), piezoelectric crystals (see published PCT international application No. wo 2003/089,138, Davis et al), particle deflection (see U.S. patent No.6,802,489, Marr et al), or electrochemically formed bubbles within the channel (see published PCT international application No. wo 2003/046,256, Hua et al). One-way valves or "check valves" may also be formed in the microchannel by spheres, flaps or membranes (see U.S. patent No.6,817,373 to Cox et al; U.S. patent No.6,554,591 to Dai et al; published PCT international application No. wo 2002/053,290, Jeon et al). Rotary-type switching valves are used for complex reactions (see published PCT international application No. wo2002/055,188, Powell et al).

Micron-sized mixing and separation of components is required to facilitate the reaction and evaluate the product. In microfluidic devices, mixing is most often carried out by diffusion in long, tortuous, variable width or turbulence-inducing channels (see U.S. patent No.6,729,352 to O' Conner et al, U.S. published patent application No.20030096310, Hansen et al). Mixing can also be achieved by electro-osmotic methods (see US patent No.6,482,306, Yager et al) or ultrasonic methods (see US patent No.5,639,423, Northrup et al). Separation in micron-scale channels typically uses 3 methods: electrophoresis, packed columns or gels within channels, or functionalization of channel walls. Electrophoresis is typically performed with charged molecules such as nucleic acids, peptides, proteins, enzymes, and antibodies, and is the simplest technique (see U.S. Pat. No.5,958,202 to Regnier et al, U.S. Pat. No.6,274,089 to Chow et al). Channel-shaped columns can be packed with beads or gels that are porous or coated with a stationary phase to facilitate separation (see published PCT international application No. wo 2003/068,402(Koehler et al), U.S. published patent application No.20020164816(Quake et al), U.S. patent No.6,814,859(Koehler et al)). Possible filler materials include silicates, talc, fuller's earth, glass wool, charcoal, activated carbon, celi plastics, silica gel, alumina, paper, cellulose, starch, magnesium silicate, calcium sulfate, silicic acid, magnesium silicate support, magnesium oxide, polystyrene, p-aminobenzyl cellulose, polytetrafluoroethylene resins, polystyrene resins, SEPHADEX^TM(Amersham Biosciences Corp., Piscataway, N.J.), SEPHAROSE^TM(Amersham Biosciences, Corp., Piscataway, N.J.), controlled pore glass beads, agarose, other solid resins known to those skilled in the art, and combinations of two or more of any of the foregoing. Magnetizable materials, such as iron oxide, nickel oxide, barium ferrite, or ferrous oxide, can also be embedded, encapsulated, or otherwise incorporated into the solid phase filler material.

The walls of the microfluidic chamber can also be functionalized with various ligands that can interact or bind to the analyte or to a contaminant in the analyte solution. Such ligands include: hydrophilic or hydrophobic small molecules, steroids, hormones, fatty acids, polymers, RNA, DNA, PNA, amino acids, peptides, proteins (including antibody binding proteins such as G-protein), antibodies or antibody fragments (FAB, etc.), antigens, enzymes, carbohydrates (including glycoproteins or glycolipids), lectins, cell surface receptors (or portions thereof), substances with positive or negative charges, and the like (see U.S. published patent application No.20040053237(Liu et al), published international PCT application No. wo 2004/007,582(Augustine et al), U.S. published patent application No.20030190608 (Blackburn)).

Thus, in some embodiments, the presently disclosed subject matter describes methods of flowing a material in a PFPE-based microfluidic device and/or mixing two or more materials in a PFPE-based microfluidic device. In some embodiments, the presently disclosed subject matter describes methods of performing chemical reactions, including but not limited to the synthesis of biopolymers, such as DNA. In some embodiments, the presently disclosed subject matter describes methods of screening samples for a characteristic. In some embodiments, the presently disclosed subject matter describes a method of dispensing material. In some embodiments, the presently disclosed subject matter describes methods of separating materials.

Method for flowing a material in a PFPE-based microfluidic device and/or mixing two materials in a PFPE-based microfluidic device

Referring now to fig. 8, a schematic diagram of the microfluidic device of the presently disclosed subject matter is shown. The microfluidic device is denoted 800. Microfluidic device 800 includes a patterned layer 802, and a plurality of wells 810A, 810B, 810C, and 810D. These apertures can be further described as an inlet aperture 810A, an inlet aperture 810B, and inlet aperture 810C, and an outlet aperture 810D. Each of the apertures 810A, 810B, 810C, and 810D is covered by a seal 820A, 820B, 820C, and 820D, which are preferably reversible seals. Seals 820A, 820B, 820C, and 820D are provided to enable materials including, but not limited to, solvents, chemical reagents, components of biochemical systems, samples, inks, reaction products, and/or solvents, chemical reagents, components of biochemical systems, samples, inks, mixtures of reaction products, and combinations thereof, to be stored, transported, or maintained in microfluidic device 800 as desired in microfluidic device 800. The seals 820A, 820B, 820C, and 820D can be reversible, i.e., detachable, so that the microfluidic device 800 can be used in chemical reactions or for other uses, and then can be sealed as desired.

With continued reference to fig. 8, in some embodiments, the wells 810A, 810B, and 810C further comprise pressure-actuated valves (including intersecting, overlapping flow channels, not shown) that can be actuated to seal the microfluidic channels associated with the wells.

With continued reference to fig. 8, the patterned layer 802 of the microfluidic device 800 includes an integrated network 830 of micron-sized channels. Optionally, the patterning layer 802 includes a functionalized surface, such as the surface shown in fig. 5A. The integrated network 830 can include a series of fluidly connected micron-scale channels designated by the following reference characters: 831, 832, 833, 834, 835, 836, 837, 838, 839 and 840. Thus, the inlet aperture 810A is in fluid communication with the micron-scale passage 831, which extends away from the aperture 810A and is in fluid communication with the micron-scale passage 832 via a bend. In the integrated network 830 depicted in fig. 8, a series of 90 degree bends are shown for convenience. However, it should be noted that the passageways and bends provided in the channels of the integrated network 830 can comprise any desired configuration, angle, or other characteristic (such as, but not limited to, segments of coiled tubing). In fact, if desired, fluid reservoirs 850A and 850B can be provided along the micron-sized channels 831, 832, 833 and 834, respectively. As shown in fig. 8, the fluid reservoirs 850A and 850B include at least one dimension that is larger than the dimension of the channels immediately adjacent thereto.

Then, with continued reference to FIG. 8, the microscale channels 832 and 834 intersect at an intersection 860A and extend into a single microscale channel 835. The micron-sized channels 835 extend into the chamber 870, which in the embodiment shown in fig. 8 has a wider dimension than the micron-sized channels 835. In some embodiments, chamber 870 comprises a reaction chamber. In some embodiments, chamber 870 comprises a mixing zone. In some embodiments, chamber 870 includes a separation region. In some embodiments, the separation region comprises a given dimension, e.g., length, of the channel, wherein the material is separated by charge, or mass, or a combination thereof, or any other physical property, wherein separation can occur at the given dimension. In some embodiments, the separation region includes an active material 880. It will be understood by those skilled in the art that the term "active material" is used herein for convenience and does not imply that the material must be activated for its intended purpose. In some embodiments, the active material comprises a chromatographic material. In some embodiments, the active material comprises a target material.

With continued reference to fig. 8, it is noted that the chamber 870 need not necessarily have dimensions wider than adjacent microscale channels. Indeed, chamber 870 can simply comprise a given segment of micron-sized channels in which at least two materials are separated, mixed, and/or reacted. Extending from the chamber 870 substantially opposite the microscale channels 835 are microscale channels 836. The microscale channel 836 forms a T-shaped connection with microscale channel 837, which extends away from aperture 810C and is in fluid communication with aperture 810C. Thus, the junction 860B is formed where the micron-sized channels 836 and 837 meet. The microscale channels 838 extend from the intersection 860B and to the fluid reservoir 850C in a direction substantially opposite the microscale channels 837. The fluid reservoir 850C is wider in size than the micron-sized channels 838 over a predetermined length. However, as described above, a given segment of the microscale channel can serve as a fluid reservoir without changing the dimensions of this segment of the microscale channel. In addition, the micro-scale channels 838 can function as reaction chambers in that a reagent flowing from the micro-scale channels 837 to the junction 860B reacts with a reagent moving from the micro-scale channels 836 to the junction 860B and enters the micro-scale channels 838.

With continued reference to fig. 8, the microscale channel 839 extends from the fluid reservoir 850C and through the bends into the microscale channel 840, substantially in the opposite direction to the microfluidic channel 838. The micro-scale channels 840 are fluidly connected to the outlet holes 810D. The outlet aperture 810D can optionally be reversibly sealed via a seal 820D, as discussed above. Again, for embodiments in which a reaction product is formed in the microfluidic device 800 and the reaction product is desired to be transported to another location in the microfluidic device 800, reversible sealing of the exit orifice 810D is desirable.

The flow of material can be directed through an integrated network 830 of micron-sized channels, including channels, fluid reservoirs, and reaction chambers, using pressure-actuated valves and similar devices known in the art, such as those described in Unger et al, U.S. patent No.6,408,878, which is hereby incorporated by reference in its entirety. The presently disclosed subject matter thus provides methods of flowing materials through a PFPE-based microfluidic device. In some embodiments, the method comprises: providing a microfluidic device comprising (i) a perfluoropolyether (PFPE) material having properties selected from the group consisting of: a viscosity of greater than about 100 centistokes (cSt); a viscosity of less than about 100cSt, with the proviso that the liquid PFPE precursor material having a viscosity of less than 100cSt is not a free radical photo-curable PFPE material; (ii) a functionalized PFPE material; (iii) a fluoroolefin-based elastomer; and (iv) combinations thereof, and wherein the microfluidic device comprises one or more microscale channels; and flowing the material in the micron-scale channels.

Also provided are methods of mixing two or more materials. In some embodiments, the method comprises: providing a microscale device that includes (i) a perfluoropolyether (PFPE) material having properties selected from the group consisting of: a viscosity of greater than about 100 centistokes (cSt); a viscosity of less than about 100cSt, with the proviso that the liquid PFPE precursor material having a viscosity of less than 100cSt is not a free radical photo-curable PFPE material; (ii) a functionalized PFPE material; (iii) a fluoroolefin-based elastomer; and (iv) combinations thereof; and contacting the first material and the second material in the apparatus to mix the first and second materials. Optionally, the micro-scale device is selected from a microfluidic device and a microtiter plate.

In some embodiments, the method comprises placing a material in the microfluidic device. In some embodiments, as best shown in fig. 10 and discussed in more detail below, the method includes applying a driving force to move the material along the microscale channel.

In some embodiments, the layer of PFPE material covers a surface of at least one of the one or more microscale channels. Optionally, the layer of PFPE material comprises a functionalized surface. In some embodiments, the microfluidic device comprises one or more patterned layers of PFPE material, and the one or more patterned layers of PFPE material define one or more microscale channels. In which case the patterned layer of PFPE can comprise a functionalized surface. In some embodiments, the microfluidic device can further comprise a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of PFPE material. See fig. 2.

In some embodiments, the method comprises at least one valve. In some embodiments the valve is a pressure-actuated valve, wherein the pressure-actuated valve is defined by one of: (a) a micron-scale channel; and (b) at least one of the plurality of holes. In some embodiments, the pressure actuated valve is actuated by introducing pressurized fluid to one of: (a) a micron-scale channel; and (b) at least one of the plurality of holes.

In some embodiments, the pressurized fluid has a pressure between about 10psi and about 40 psi. In some embodiments, the pressure is about 25 psi. In some embodiments, the material comprises a fluid. In some embodiments, the material fluid comprises a solvent. In some embodiments, the solvent comprises an organic solvent. In some embodiments, the material flows in a predetermined direction along the microscale channel.

For the case of mixing two materials, which in some embodiments can include mixing two reactants to perform a chemical reaction, contacting the first material and the second material is performed in a mixing zone defined in one or more microscale channels. The mixing zone can include a geometry selected from the group consisting of a T-junction, a coil, a long channel, a micron-scale chamber, and a constriction. Optionally, the first material and the second material are disposed in separate channels of the microfluidic device. In addition, the contacting of the first material and the second material can be performed in a mixing zone defined by the intersection of the channels.

For the mixing method, the method can include flowing the first material and the second material along a predetermined direction in a microfluidic device, and can include flowing the mixed material along the predetermined direction in the microfluidic device. In some embodiments, the mixed material can be contacted with a third material to form a second mixed material. In some embodiments the mixed material includes a reaction product, and the reaction product is subsequently reacted with a third reagent. One skilled in the art, upon reading the presently disclosed subject matter, will recognize that the description of the mixing method provided immediately above is for purposes of illustration and is not meant to be limiting. Thus, the presently disclosed methods of mixing materials can be used to mix a plurality of materials and form a plurality of mixed materials and/or a plurality of reaction products. The mixed material, including but not limited to reaction products, can flow into the exit orifice of the microfluidic device. A driving force can be applied to move the material through the microfluidic device. See fig. 10. In some embodiments the mixed material is recycled.

In one embodiment, a microtiter plate is used, which can comprise one or more wells. In some embodiments, the layer of PFPE material covers a surface of at least one hole of the one or more holes. The layer of PFPE material can include a functionalized surface. See fig. 5B.

Method for synthesizing biopolymer in PFPE-based microfluid equipment

In some embodiments, the presently disclosed PFPE-based microfluidic devices can be used for biopolymer synthesis, e.g., for synthesizing oligonucleotides, proteins, peptides, DNA, and the like. In some embodiments, such biopolymer synthesis systems include integrated systems comprising: an array of reservoirs, fluidic logic for selecting flows from the distinct reservoirs, an array of channels, reservoirs, and reaction chambers in which synthesis is performed, and fluidic logic that determines which channel the selected reagent flows into.

Referring now to fig. 9, a plurality of reservoirs, for example, reservoirs 910A, 910B, 910C, and 910D, have bases A, C, T and G, respectively, located therein. Four flow channels 920A, 920B, 920C and 920D are connected to reservoirs 910A, 910B, 910C and 910D. Four control passages 922A, 922B, 922C, and 922D (shown in phantom) are positioned across, wherein control passage 922A permits flow only through flow passage 920A (i.e., sealing flow passages 920B, 920C, and 920D) when control passage 922A is pressurized. Similarly, when boosted, control passage 922B allows flow only through passage 920B. Likewise, selective pressurization of control channels 922A, 922B, 922C, and 922D sequentially selects a desired seat A, C, T and G for a desired reservoir 910A, 910B, 910C, or 910D. The fluid then passes through flow channel 920E into a multi-channel flow controller 930, (including, for example, any of the systems shown in fig. 8) which, in turn, directs the fluid to flow into one or more of a plurality of synthesis channels or reaction chambers 940A, 940B, 940C, 940D, or 940E, where solid phase synthesis is performed.

In some embodiments, instead of beginning with the desired bases a, C, T, and G, an agent selected from one of a nucleotide and a polynucleotide is placed in at least one of reservoirs 910A, 910B, 910C, and 910D. In some embodiments, the reaction product comprises a polynucleotide. In some embodiments, the polynucleotide is DNA.

Accordingly, upon reading this disclosure, one skilled in the art will recognize that the presently disclosed PFPE-based microfluidic devices may be used to synthesize biopolymers, as described in U.S. patent No.6,408,878 to Unger et al and U.S. patent 6,729,352 to O' Conner et al, and/or as described in a combinatorial synthesis system as described in U.S. patent No.6,508,988 to van Dam et al, each of which is incorporated herein by reference in its entirety.

A method of introducing a PFPE-based microfluidic device into an integrated fluid flow system.

In some embodiments, a method of performing a chemical reaction or flowing a material within a PFPE-based microfluidic device comprises introducing the microfluidic device into an integrated fluid flow system. Referring now to fig. 10, a system for performing a method of flowing a material in a microfluidic device and/or performing a chemical reaction in accordance with the presently disclosed subject matter is diagrammatically depicted. The system itself is generally designated 1000. The system 1000 can include a central processing unit 1002, one or more driving force actuators 1010A, 1010B, 1010C, and 1010D, a collector 1020, and a detector 1030. In some embodiments, detector 1030 is in fluid communication with a microfluidic device (shown in phantom). The system of fig. 8 the microfluidic device 1000 and these reference numbers of fig. 8 can be used for fig. 10. The Central Processing Unit (CPU)1002 can be, for example, a general purpose personal computer with an associated monitor, keyboard, or other desired user interface. The driving force actuators 1010A, 1010B, 1010C, and 1010D can be any suitable driving force actuator as understood by one of ordinary skill in the art after reading the presently disclosed subject matter. For example, the driving force actuators 1010A, 1010B, 1010C, and 1010D can be pumps, electrodes, syringes (injectors), syringes (syringes), or other such devices that can be used to force material through a microfluidic device. Representative driving forces themselves therefore include capillary action, pump-driven fluid flow, electrophoretic-type fluid flow, pH gradient-driven fluid flow, or other gradient-driven fluid flow.

The driving force actuator 1010D is shown in connection with the outlet aperture 810D in the schematic of fig. 10, and will be described below to illustrate that at least a portion of this driving force can be provided at the end of a desired flow of solution, reagent, and the like. A collector 1020 is also provided to indicate that reaction product 1048 (discussed below) can be collected at the end of the system flow. In some embodiments, the collector 1020 comprises a fluid reservoir. In some embodiments, collector 1020 comprises a substrate (substrate). In some embodiments, collector 1020 comprises a detector. In some embodiments, the collector 1020 includes a target of a treatment requiring treatment. For convenience, the system flow is generally represented in fig. 10 by directional arrows F1, F2, and F3.

With continued reference to fig. 10, in some embodiments chemical reactions are performed in an integrated flow system 1000. In some embodiments, a material 1040, e.g., a chemical reagent, is introduced into the microfluidic device 1000 through the aperture 810A, while a second material 1042, e.g., a second chemical reagent, is introduced into the microfluidic device 1000 via the inlet aperture 810B. Optionally, the microfluidic device 1000 comprises a functionalized surface (see fig. 5A). The driving force actuators 1010A and 1010B propel chemical reagents 1040 and 1042 into microfluidic channels 831 and 833, respectively. Chemical reagents 1040 and 1042 continue to flow into fluid reservoirs 850A and 850B, where the stored reagents 1040 and 1042 are collected. Chemical reagents 1040 and 1042 continuously flow into microfluidic channels 832 and 834 to intersection 860A, where initial contact between chemical reagents 1040 and 1042 is made. The chemical reagents 1040 and 1042 then continue to flow into the reaction chamber 870, where the chemical reaction between the chemical reagents 1040 and 1042 proceeds.

With continued reference to fig. 10, the reaction product 1044 flows into the microscale channel 836 and then to the intersection 860B. Chemical 1046 then begins reacting with reaction product 1044 at intersection 860B, through reaction chamber 838 and into fluid reservoir 850C. Forming second reaction product 1048. Second reaction product 1048 flows continuously through micron-scale passage 840 to well 810D and finally into collector 1020. It is therefore noted that CPU 1002 actuates drive force actuator 1010C such that chemical 1046 is released at the appropriate time to contact reaction product 1044 at intersection 860B.

Representative applications of x.d. microfluidic devices

In some embodiments, the presently disclosed subject matter discloses methods of screening samples for a property. In some embodiments, the presently disclosed subject matter discloses a method of dispensing a material. In some embodiments, the presently disclosed subject matter discloses methods of separating materials. Accordingly, one of ordinary skill in the art will recognize that the microfluidic devices described herein can be used in a number of applications, including, but not limited to, genomic profiling, rapid separation, sensors, nanoscale reactions, inkjet printing, drug delivery, lab-on-a-chip, in vitro diagnostics, injection nozzles, biological research, high throughput screening techniques, such as for drug discovery and materials science, diagnostic and therapeutic tools, research tools, and biochemical monitoring of food and natural resources such as soil, water and/or air samples collected with portable or stationary monitoring devices.

Method for screening samples for characteristics

In some embodiments, the presently disclosed subject matter discloses methods of screening samples for a property. In some embodiments, the method comprises:

(a) providing a microscale device that includes:

(i) a perfluoropolyether (PFPE) material having properties selected from the group consisting of: a viscosity of greater than about 100 centistokes (cSt); and a viscosity of less than about 100cSt, with the proviso that the liquid PFPE precursor material having a viscosity of less than 100cSt is not a free radical photo-curable PFPE material;

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof;

(b) providing a target material;

(c) placing the sample in a micron-scale apparatus;

(d) contacting the sample with a target material; and

(e) detecting an interaction between the sample and the target, wherein the presence or absence of the interaction is indicative of a characteristic of the sample.

Referring again to fig. 10, at least one of the materials 1040 and 1042 includes a sample. In some embodiments, at least one of materials 1040 and 1042 comprises a target material. Thus, "sample" generally refers to any material in which information about their properties is desired. Meanwhile, "target material" refers to any material that can be used to provide information related to the characteristics of a sample based on the interaction between the target material and the sample. In some embodiments, for example, the interaction occurs when sample 1040 contacts target material 1042. In some embodiments, the interaction produces reaction product 1044. In some embodiments, the interaction comprises a binding interaction (binding event). In some embodiments, the binding interaction includes, for example, an interaction between an antibody and an antigen, between an enzyme and a substrate, or more specifically, between a receptor and a ligand, or between a catalyst and one or more chemical agents. In some embodiments, the reaction product is detected by detector 1030.

In some embodiments, the method includes placing the target material in at least one of a plurality of channels. Referring again to fig. 10, in some embodiments, the target material includes an active material 880. In some embodiments, the target material, the sample, or both the target material and the sample are bound to a functionalized surface. In some embodiments, the target material comprises a substrate, such as an unpatterned layer. In some embodiments, the substrate comprises a semiconductor material. In some embodiments, at least one of the plurality of channels of the microfluidic device is in fluid communication with a substrate, e.g., an unpatterned layer. In some embodiments, the target material is disposed on a substrate, such as a non-patterned layer. In some embodiments, at least one of the one or more channels of the microfluidic device is in fluid communication with a target material disposed on the substrate.

In some embodiments, the method comprises placing a plurality of samples in at least one of the plurality of channels. In some embodiments, the sample is selected from the group consisting of therapeutic agents, diagnostic agents, research reagents, catalysts, metal ligands, non-biological organic materials, inorganic materials, food, soil, water, and air. In some embodiments, the sample comprises one or more constituent members of one or more libraries of chemical or biological compounds or components. In some embodiments, the sample comprises one or more of a nucleic acid template (a nucleic acid template), a sequence reagent (a sequencing reagent), a primer (a primer), a primer extension product, a restriction enzyme (restriction enzyme), a PCR reagent, a PCR reaction product, or a combination thereof. In some embodiments, the sample comprises one or more of an antibody, a cellular receptor, an antigen, a receptor ligand, an enzyme, a substrate, an immunochemical, an immunoglobulin, a virus binding component, a protein, a cytokine, a growth factor, an inhibitor, or a combination thereof.

In some embodiments, the target material comprises one or more of an antigen, an antibody, an enzyme, a restriction enzyme, a dye, a fluorescent dye, a sequence reagent, a PCR reagent, a primer, a receptor, a ligand, a chemical reagent, or a combination thereof.

In some embodiments, the interaction comprises a binding interaction. In some embodiments, the detection of the interaction is by at least one or more of a spectrophotometer, fluorometer, photodiode, photomultiplier tube, microscope, scintillation counter, camera, CCD camera, membrane, optical detection system, temperature sensor, conductivity meter, potentiometer, ammeter, pH meter, or combinations thereof.

Accordingly, after reading this disclosure, one of ordinary skill in the art will recognize that the presently disclosed PFPE-based microfluidic devices may be used in a variety of screening techniques, such as those described in U.S. patent No 6,749,814 issued to Bergh et al, 6,737,026 issued to Bergh et al, 6,630,353 issued to park et al, 6,620,625 issued to Wolk et al, 6,558,944 issued to park et al, 6,547,941 issued to Kopf-silk et al, 6,529,835 issued to Wada et al, 6,495,369 issued to Kercso et al, and 6,150,180 issued to park et al, each of which is incorporated herein by reference in its entirety. Further, after reading this disclosure, one of ordinary skill in the art will recognize that the presently disclosed PFPE-based microfluidic devices can be used, for example, to detect DNA, proteins, or other molecules associated with a particular biochemical system, as described in US patent No.6,767,706 to Quake et al, which is incorporated herein by reference in its entirety.

Method for dispensing material

Additionally, the presently disclosed subject matter describes a method of dispensing material. In some embodiments, the method comprises:

(a) providing a microfluidic device comprising:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof; and wherein the microfluidic device comprises one or more micron-scale channels, and wherein at least one of the one or more micron-scale channels comprises an exit orifice;

(b) providing at least one material;

(c) placing at least one material in at least one of the one or more microscale channels; and

(d) at least one material is dispensed through the outlet orifice.

In some embodiments, the layer of PFPE material covers a surface of at least one of the one or more microscale channels.

Referring again to fig. 10, in some embodiments, materials, e.g., material 1040, second material 1042, chemical 1046, reaction product 1044, and/or reaction product 1048, flow through outlet aperture 810D and are dispensed in or on collector 1020. In some embodiments, the target material, the sample, or both the target and the sample are bound to a functionalized surface.

In some embodiments, the material comprises a drug. In some embodiments, the method comprises metering in a predetermined dose of the drug. In some embodiments, the method comprises dispensing a predetermined dose of the drug.

In some embodiments, the material comprises an ink composition. In some embodiments, the method includes dispensing an ink composition on a substrate. In some embodiments, the dispensing of the ink composition on the substrate forms a printed image.

Accordingly, after reading this disclosure, one of ordinary skill in the art will recognize that the presently disclosed PFPE-based microfluidic devices can be used in microfluidic printing as described in US patent No 6,334,676 to kaszzzuk et al, 6,128,022 to DeBoer et al, and 6,091,433 to Wen, each of which is incorporated herein by reference in its entirety.

Method for separating materials by X.D.3

In some embodiments, the presently disclosed subject matter describes a method of separating materials, the method comprising:

(a) providing a microfluidic device comprising:

(ii) A functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof; and wherein the microfluidic device comprises one or more microscale channels, wherein at least one of the one or more microscale channels comprises a separation region;

(b) placing a mixture comprising at least a first material and a second material in the microfluidic device;

(c) flowing the mixture through the separation zone; and

(d) the first material is separated from the second material in the separation region to form at least one separated material.

Referring again to fig. 10, in some embodiments, at least one of material 1040 and second material 1042 comprises a mixture. For example, material 1040, e.g., a mixture, flows through the microfluidic system into chamber 870, which in some embodiments includes a separation region. In some embodiments, the separation region includes an active material 880, e.g., a chromatographic material. The material 1040, e.g., mixture, is separated in the chamber 870 (e.g., separation chamber) to form a third material 1044, e.g., a separated material. In some embodiments, the separated material 1044 is detected by detector 1030.

In some embodiments, the separation region comprises chromatographic material. In some embodiments, the chromatographic material is selected from size-separation matrix (size-separation matrix), affinity-separation matrix (affinity-separation matrix), and gel-exclusion matrix (gel-exclusion matrix), or a combination thereof.

In some embodiments, the first or second material comprises one or more constituent members of one or more libraries of chemical or biological compounds or components. In some embodiments, the first or second material comprises one or more of a nucleic acid template, a sequence reagent, a primer extension product, a restriction enzyme, a PCR reagent, a PCR reaction product, or a combination thereof. In some embodiments, the first or second material comprises one or more of an antibody, a cellular receptor, an antigen, a receptor ligand, an enzyme, a substrate, an immunochemical, an immunoglobulin, a virus-binding component, a protein, a cytokine, a growth factor, an inhibitor, or a combination thereof.

In some embodiments, the method comprises detecting the separated material. In some embodiments, the detection of the separated material is performed by at least one or more of a spectrophotometer, fluorometer, photodiode, photomultiplier tube, microscope, scintillation counter, camera, CCD camera, membrane, optical detection system, temperature sensor, conductivity meter, potentiometer, ammeter, pH meter, or combinations thereof.

Accordingly, after reading this disclosure, one of ordinary skill in the art will recognize that the presently disclosed PFPE-based microfluidic devices may be used to separate a variety of materials, as described in US patent No 6,752,922 issued to Huang et al, US6,274,089 issued to Chow et al, and US6,444,461 issued to Knapp et al, each of which is incorporated herein by reference in its entirety.

Application of functionalized microfluidic devices

Fluid microchip technology is increasingly being used as an alternative to traditional chemical and biological laboratory functions. Microchips have been manufactured to perform complex chemical reactions, separations, and detections on a single device. These "lab-on-a-chip" applications facilitate fluid and analyte transport, with the advantage of reduced time and chemical consumption and ease of automation.

Various biochemical analyses, reactions, and separations have been performed within microchannel systems. High throughput screening assays for synthetic molecules and natural products are of great interest. Microfluidic devices for screening various molecules based on their ability to inhibit the interaction between an enzyme and a fluorescently labeled substrate have been described (U.S. patent No.6,046,056 to Parse et al). Such devices can screen natural or synthetic libraries of potential drugs for their antagonistic or agonistic properties, as described by Parse et al. Types of molecules that can be screened include, but are not limited to, small organic or inorganic molecules, polysaccharides, peptides, proteins, nucleic acids or extracts of biological materials such as bacteria, fungi, yeast, plant and animal cells. The analyte compound can remain free in solution or attached to a solid support such as agarose, cellulose, dextran, polystyrene, carboxymethylcellulose, polyethylene glycol (PEG), filter paper, nitrocellulose, ion exchange resins, plastic membranes, glass beads, polyamine methyl vinyl ether maleic acid copolymers, amino acid copolymers, ethylene maleic acid copolymers, nylon, silk, and the like. The compounds can be tested as pure compounds or in a cell. For example, U.S. patent No.6,007,690 issued to Nelson et al relates to microfluidic molecular diagnostics for the purification of DNA from a whole blood sample. The apparatus uses an enrichment channel that cleans or concentrates the analyte sample. For example, the enrichment channel can accommodate antibody-coated beads for removal of various cellular fractions using their antigenic components, or can accommodate chromatographic components such as ion exchange resins or hydrophobic or hydrophilic membranes. The device can also include a reactor chamber in which various reactions can be performed on the analyte, such as labeled reactions or digestion reactions in the case of protein analytes. Furthermore, US published patent application No.20040256570(Beebe et al) describes a device in which the interaction between an antibody and antigenic analyte material coated on the exterior of a liposome is detected when the interaction causes the liposome to lyse and the liposome releases detectable molecules. U.S. published patent application of Miller et al No.20040132166 provides for sensing environmental factors such as pH, humidity and O critical to cell growth₂Horizontal microfluidic devices. The reaction chambers in these devices can be used as bioreactors for growing cells, allowing them to be used to transfect cells with DNA and produce proteins, or to test the potential bioavailability of a drug substance by measuring its uptake rate through the CACO-2 cell layer.

In addition to growing cells, microfluidic devices have also been used to sort (sort) cells. U.S. patent No.6,592,821 to Wada et al describes a fluid dynamic focusing on sorting cellular and subcellular components (including single molecules such as nucleic acids, polypeptides or other organic molecules, or larger cellular components such as organelles). This method can be used to sort out cell viability or other cell expression functions.

Amplification, separation, sequencing and identification of nucleic acids and proteins are common microfluidic device applications. For example, U.S. Pat. No.5,939,291 to Loewy et al describes a microfluidic device for isothermal nucleic acid amplification using electrostatic techniques. The apparatus can be used in conjunction with a number of common amplification reaction strategies including PCR (polymerase chain reaction), LCR (ligase chain reaction), SDA (strand displacement amplification), NASBA (nucleic acid sequence based amplification), and TMA (amplification of transcription media). US patent No.5,993,611 to Moroney et al describes an apparatus for analyzing, amplifying or otherwise manipulating nucleic acids using capacitive charging. Devices have been designed that can sort DNA by size and analyze restriction fragment length polymorphisms (see Quake et al, U.S. Pat. No.6,833,242). The device can also find particular use in forensic applications, such as DNA fingerprinting. U.S. patent No.6,447,724 to Jensen et al describes a microfluidic device that is capable of identifying components of a mixture based on the different fluorescence lifetimes of labels attached to the individual constituent members of the mixture. Such devices can be used to analyze the sequence response of nucleic acids, proteins or oligosaccharides or to examine or query the constituent members of a combinatorial library of organic molecules.

Other microfluidic devices that focus on specific protein applications include devices that promote protein crystal growth in microfluidic channels (see U.S. Pat. No.6,409,832 to Weigl et al). In this apparatus, a protein sample and a solvent are directed into a channel having laminar flow characteristics forming diffusion sections that provide well-defined crystallization. U.S. published patent application No.2004/0121449 to Pugia et al describes an apparatus capable of separating red blood cells from plasma using minimal centrifugal force at sample sizes as small as 5 microliters. The apparatus is particularly useful in clinical diagnostics and also for separating any particulate matter from a liquid.

As described in part above, microfluidic devices have been used as microreactors for a variety of chemical and biological applications. The chambers in these devices can be used for sequencing, restriction enzyme digestion, Restriction Fragment Length Polymorphism (RFLP) analysis, nucleic acid amplification, or gel electrophoresis (see U.S. Pat. No.6,130,098 to Handique et al). Many chemical titration reactions can be carried out in equipment (see U.S. published patent application No.20040258571, Lee et al), including acid-based or precipitation-based titrations (e.g., Ag (I) with Cl)^-，Br^-，I^-Or SCN^-Titration), complex formation (e.g., Ag (I) with CN)^-) Or redox reactions (e.g., Fe (II)/Fe (III) and Ce (III)/Ce (IV)). Furthermore, sensors for potentiometry, amperometry, spectrophotometry, turbidimetry, fluorimetry or calorimetry can be attached to the device. Protein fractionation (fractionation) (see U.S. published patent application No.20040245102, Gilbert et al) type physical or biological properties can be used for protein expression analysis (finding molecular markers, determining molecular basis or distribution causing disease or interpreting protein structure/functional group interrelationships). Various electrophoretic techniques, including capillary isoelectric focusing, capillary zone electrophoresis, and capillary gel electrophoresis, have been used in microfluidic devices for fractionating proteins (see U.S. Pat. No.6,818,112 to Schneider et al). Different electrophoretic techniques can be used in series, with or withoutThere is no labeling step to aid in quantitation and use in conjunction with various elution techniques (e.g., hydrodynamic salt mobilization, pH mobilization, or electroosmotic flow) to further separate proteins. Various other materials have been used to assist in the separation process in microfluidic devices. Such materials may be attached to the walls of a channel in the device or present as a separate matrix within the channel (see Paul, U.S. Pat. No.6,581,441; Wada et al, U.S. Pat. No.6,613,581). Parallel separation channels can be present to separate many samples simultaneously. The solid separation medium can exist as discrete particles or as a monolithic solid with pores. Possible materials include silica gel, agarose based gel, polyacrylamide gel, colloidal solutions such as gelatin, starch, nonionic macroreticular and macroporous resins (e.g. AMBERCHROM)^TM(Rohm and Haas Co, Philadelphia, Pa., USA), AMBERLITE^TM(Rohm and Haas Co, Philadelphia, Pa.), DOWEX^TM(The Dow Chemical Company, Midland, Mich., USA),(Rohm and Haas Co, Philadelphia, Pa., USA), and the like), or a material that exists as beads (glass, metal, silica, acrylic, SEPHAROSE)^TMCellulose, ceramic, polymer, and the like). These materials can also present various biologically significant molecules on their surfaces to aid in separation (e.g., lectins can be bound to carbohydrates and antibodies can be bound to antigenic groups on different proteins). Membranes within microchannels have been used for electro-osmotic separations (see U.S. patent No.6,406,605 to Moles). Suitable membranes can be composed of materials such as track etched polycarbonate or polyimide.

Temperature, concentration and flow gradients have also been used to assist separation in microfluidic devices. U.S. published patent application No.20040142411(Kirk et al) discloses the use of chemotaxis (cell motility induced by concentration gradients of soluble chemotactic stimuli), chemotaxis (cell motility in response to concentration gradients of matrix-bound stimuli), and chemoinvasiveness (cell motility into and/or through a barrier or gel matrix in response to stimuli). Chemotactic stimuli include chemical repellants (chemorepellents) and chemical attractants (chemoattractants). A chemical attractant is any substance that attracts cells. Examples include, but are not limited to, hormones such as epinephrine and antidiuretic hormone; immunological reagents such as interleukin-2 (interleukin-2); growth factors, chemokines, cytokines, and various peptides, small molecules, and cells. Chemical repellents include irritants (irritants), such as benzalkonium chloride, propylene glycol, methanol, acetone, sodium lauryl sulfate, hydrogen peroxide, 1-butanol, ethanol, and dimethyl sulfoxide; toxins, such as cyanide, carbonyl cyanide, chlorophenylhydrazone (chlorophenylhydrozone); endotoxins and bacterial lipopolysaccharides; a virus; pathogenic bacteria; and a pyrogen. Non-limiting examples of cells that can be manipulated by these techniques include lymphocytes, monocytes, leukocytes, macrophages, mast cells, T cells, B cells, neutrophils, basophils, fibroblasts, tumor cells and many other cells.

Microfluidic devices as sensors have attracted attention in recent years. Such microfluidic sensors can include dye-based detection systems, affinity-based detection systems, microfabricated weight analyzers, CCD cameras, photodetectors, optical microscopy systems, electrical systems, thermocouples, temperature-dependent resistors, and pressure sensors. Such devices have been used to detect biomolecules (see published PCT International application No. WO 2004/094,986, Althaus et al), including polynucleotides, proteins and viruses, by their interaction with probe molecules capable of providing an electrochemical signal. For example, insertion of a probe molecule for nucleic acid samples, such as doxorubicin (doxorubicin), can reduce the amount of free doxorubicin that comes into contact with the electrode; and variations in the electrical signal results. Devices have been described which contain means for detecting and controlling the deviceEnvironmental factors inside the reaction chamber such as humidity, pH, dissolved O₂And dissolved CO₂The sensor of (see published PCT International application No. WO 2004/069,983, Rodgers et al). Such devices are particularly useful for growing and maintaining cells. The carbon content of the sample can be measured in an apparatus (see U.S. Pat. No.6,444,474 to Thomas et al) in which ultraviolet radiation oxidizes organics to CO₂It is quantitatively analyzed by conductivity measurement or infrared method. Capacitive sensors for microfluidic devices (see published PCT international application No. wo 2004/085,063, Xie et al) can be used to measure pressure, flow, fluid level and ion concentration.

Another application of microfluidic systems involves high throughput injection of cells (see published PCT International application No. WO 00/20554, Garman et al). In such devices, cells are pushed into an injection needle so that they can be injected with a variety of materials including molecules and macromolecules, genes, chromosomes, or organelles. The device may also be used to extract substances from cells and may be used in various fields such as gene therapy, pharmaceutical or agrochemical research, and diagnostics. Microfluidic devices have also been used as ink delivery devices in ink jet printing (see US patent No.6,575,562, Anderson et al), and to direct sample solutions to electrospray ionization tips for mass spectrometry (see US patent No.6,803,568, bouse et al). Transdermal drug delivery systems have also been reported (see published PCT International application No. WO2002/094,368, Cormier et al), as well as devices containing light altering elements for use in spectroscopy applications (see U.S. Pat. No.6,498,353, Nagle et al).

Use of functionalized microtiter plates

The presently disclosed materials and methods can also be used in the design and manufacture of devices used in the manner of microtiter plates. Microtiter plates have various uses in the fields of protein metabolism (proteomics), genomic (genomics) and drug discovery, environmental chemical analysis, parallel synthesis, cell culture, molecular biology and high throughput screening of immunoassays. Common base materials for microtiter plates include hydrophobic materials, such as polystyrene and polypropylene, and hydrophilic materials, such as glass. Silicon, metal, polyester, polyolefin and polytetrafluoroethylene surface layers have also been used in microtiter plates.

The surface layers can be selected based on their particular application of solvent and temperature compatibility and their ability (or lack thereof) to interact with the molecules or biomolecules to be analyzed or manipulated. Chemical modification of the base material can often be used to tailor the microtiter plate specifically to its desired function by modifying the surface characteristics or by providing sites for covalent attachment of molecules or biomolecules. The functionalizable nature of the presently disclosed materials is well suited for these purposes.

Some applications require a surface with low binding properties. Proteins and many other biomolecules (e.g., eukaryotic and microbial cells) can be passively adsorbed onto polystyrene by hydrophobic or ionic interactions. Some surface modified base materials have been developed to address this problem.Ultra Low Attachment (Corning incorporated-Life Sciences, Acton, Mass.) is a hydrogel-coated polystyrene. The hydrogel coating causes the surface to become neutral and hydrophilic, preventing the attachment of almost all cells. The containers made from this surface layer can be used to prevent serum protein uptake, to prevent the isolation of anchorage dependent cells (MDCK, VERO, C6, etc.), to selectively culture tumor or virus transformed cells as unattached colonies, to prevent stem cells from undergoing attachment-mediated mutation, and to study macrophage activation and inactivation mechanisms. NUNC MINISORP^TM(Nalgene Nunc International, Naperville, Ill.) is a polyvinyl product that has low protein affinity and can be used for DNA probe and serotype analysis where nonspecific binding is an issue.

For other applications, materials have been modified to enhance their ability to adhere to cells and other biomolecules. NUNCLONA^TM(Nalgene Nunc International) is a polystyrene surface layer that is rendered hydrophilic and negatively charged by corona or plasma discharge treatment to add surface carboxyl groups. This material has been used for cell culture of various cells. Polyolefin and polyester materials have also been treated to enhance their hydrophilicity and thus become good surfaces for cell adhesion and growth (e.g., PERMANOX)^TMAnd THERMANOX^TMAlso available from Nalgene Nunc International). The base material can be coated with poly-D-lysine, collagen or fibronectin (fibronectin) to create a positively charged surface, which can also enhance cell attachment, growth and differentiation.

In addition, other molecules can be adsorbed onto the plate body of the microtiter plate. NuncMAXISORP^TM(Nalgene Nunc) is a modified polystyrene base with high affinity for polar molecules and is recommended for use in surface layers onto which antibodies need to be adsorbed, as is the case for many ELISA assays. The surface layer can also be modified to interact with the analyte in a more specific manner. Examples of such functional modifications include modified surfaces for the quantitative analysis and detection of nickel-chelates of histidine-tagged fusion proteins and modified surfaces for the capture of glutathione of GST-tagged fusion proteins. When acting with biotinylated proteins, a surface layer coated with streptavidin may be used.

Some modified surfaces provide sites for covalent attachment of various molecules or biomolecules. COVALINK^TMThe NH secondary amine surface (Nalgene Nunc International) is a polystyrene surface covered with secondary amines, which can bind proteins and peptides via carbodiimide chemistry using their carboxyl groups or bind DNA by 5' phosphoamide bond formation (again using carbodiimide chemistry). Other molecules, carbohydrates, hormones, small molecules, etc. containing or modified to contain carboxylate groups can also be bound to the surface. The epoxy groups being covalently bondedA bond means attaches the group to another useful moiety on the surface layer. Epoxy-modified surfaces have been used to create DNA chips using the reaction of amino-modified oligonucleotides with the surface. The surface layer with immobilized oligonucleotides can be used in high throughput DNA and RNA detection systems and for automated DNA amplification applications.

Other uses of microtiter plates are to modify the surface to become more hydrophobic, to become more compatible with organic solvents or to reduce the absorption of drugs (typically small organic molecules). For example, total drug analysis tests generally rely on the use of acetonitrile to precipitate proteins and salts from plasma or serum samples. The drug to be tested must remain in solution for subsequent quantitative analysis. Organic solvent compatible microtiter plates are also useful as High Performance Liquid Chromatography (HPLC) or liquid chromatography/mass spectrometry (LC/MS) preparative equipment and as combinatorial chemistry or parallel synthesis reaction vessels (for solution-type or solid-phase-type chemistry). Examples of surfaces for these types of uses include multifhem^TMMicroplates (Whatman, Inc., Florham Park, N.J.) andsolvinert (Millipore, Billerica, Mass., USA).

Method of using functionalized perfluoropolyether networks as gas separation membranes

The presently disclosed subject matter provides for the use of functionalized perfluoropolyether (PFPE) networks as gas separation membranes. In some embodiments, the functionalized PFPE network is used as a gas separation membrane to separate a gas selected from CO₂Methane, H₂CO, CFC, CFC substitutes, organics, nitrogen, methane, H₂S, amine, fluorocarbon, fluoroolefin, and O₂. In some embodiments, the functionalized PFPE network is used to separate gases during water purification. In some embodiments, the gas separation membrane comprises a stand-alone membrane. In some embodiments, the gas separation membrane comprises a compositeAnd (3) a membrane.

In some embodiments, the gas separation membrane comprises a comonomer. In some embodiments, the comonomer modulates the permeability properties of the gas separation membrane. Furthermore, the mechanical strength and durability of such films can be fine-tuned by adding composite fillers such as silica particles and other fillers to the film. Thus, in some embodiments, the film further comprises a composite filler. In some embodiments, the composite filler comprises silica particles.

Examples

The following examples are presented to provide guidance to those of ordinary skill in the art in practicing representative embodiments of the presently disclosed subject matter. In view of this disclosure and general knowledge in the art, those skilled in the art will recognize that the following examples are intended to be illustrative, and that many changes and modifications can be made without departing from the scope of the presently disclosed subject matter.

General considerations

PFPE microfluidic devices have been previously reported by Rolland, J.et al, JACS 2004, 126, 2322-2323, the entire contents of which are incorporated herein by reference. The particular PFPE materials disclosed in Rolland, j. et al do not undergo chain extension and therefore do not have the multiple hydrogen bonds that are present when the PFPE is chain extended with a diisocyanate linker. The material also does not have a higher molecular weight between the cross-links that is required to improve mechanical properties such as modulus and tear strength that are critical for various microfluidic applications. In addition, this material is not functionalized to incorporate various moieties, such as charged species, biopolymers, or catalysts.

Various methods of addressing these issues are described herein. Included among these improvements are methods that describe chain extension, improved adhesion to PFPE layers and to other substrates such as glass, silicon, quartz and other polymers, and the ability to incorporate functionalized monomers that can alter wettability or can attach catalysts, biomolecules or other substances. Also described are improved methods of curing PFPE elastomers, which include thermal radical curing, two-part curing chemistry, and photocuring using photoacid generators.

Example 1

A liquid PFPE precursor having the structure shown below (where n ═ 2) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The slides were then placed in a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thick layer is then peeled off, trimmed, and an access hole is formed through it using a luer punch (luer stub). The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The composite layer was then placed in an oven and heated at 120 ℃ for 2 hours. The aforementioned thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a fully cured PFPE smooth layer on a glass slide and heated at 120 ℃ for 15 hours. A small needle is then placed into the inlet to introduce fluid and actuate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 2

Thermal radical mode

Glass

The liquid PFPE precursor terminated with methacrylate groups was mixed with 1 wt% 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 20 hours under a nitrogen purge. The cured layer is then peeled off, trimmed, and an access hole is formed through it using a luer punch (luer stub). The layer was then placed on a clean glass slide and fluid was introduced through the inlet holes.

Example 3

Thermal radical regime-partial cure

Layer-to-layer bonding

The liquid PFPE precursor terminated with methacrylate groups was mixed with 1 wt% 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layers were then placed on a partially cured PFPE smooth layer on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 4

Thermal radical regime-partial cure

Adhesion to polyurethane

A photocurable liquid urethane precursor containing methacrylate groups was mixed with 1 wt% 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a smooth layer of partially cured PFPE on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 5

Thermal radical regime-partial cure

Adhesion to silicone-containing polyurethanes

A photocurable liquid polyurethane precursor containing PDMS blocks and methacrylate groups was mixed with 1 wt% 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch (luer stub). The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a smooth layer of partially cured PFPE on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 6

Thermal radical regime-partial cure

Adhesion to PFPE-PDMS Block copolymers

The liquid precursor, which is end-capped with methacrylate groups and contains both PFPE and PDMS blocks, was mixed with 1 wt% of 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a smooth layer of partially cured PFPE on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 7

Thermal radical regime-partial cure

Glass bonding

The liquid PFPE precursor terminated with methacrylate groups was mixed with 1 wt% 2, 2-azobisisobutyronitrile and poured onto a microfluidic master containing channel shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The partially cured layer was peeled away from the wafer and a luer punch was used to punch the entry hole. This layer was then placed on the surface of a glass slide that had been treated with a silane coupling agent, trimethoxysilylpropyl methacrylate. The layer was then placed in an oven and heated at 65 ℃ for 20 hours to allow the PFPE layer to permanently bond to the glass slide. A small needle is then placed into the inlet to introduce the fluid.

Example 8

Thermal radical regime-partial cure

PDMS bonding

A liquid poly (dimethylsiloxane) precursor was poured onto a microfluidic master containing channel-shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. Independently, a second master containing channel-shaped 100- μm structural features spin-coated droplets of a liquid methacrylate unit-terminated PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The layer of PDMS was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. This layer was then treated with oxygen plasma for 20 minutes, followed by treatment with a silane coupling agent, trimethoxysilylpropyl methacrylate. A treated PDMS layer was placed on the surface of the thin layer of partially cured PFPE and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a smooth layer of partially cured PFPE on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 9

Thermal radical mode

Using SYLGARDAnd PDMS adhesion of functionalized PDMS.

The liquid poly (dimethylsiloxane) precursor is designed such that it may be SYLGARDOr a portion of a curing component. The precursor contains latent functional groups such as epoxy, methacrylate, or amine and is mixed with standard curing agents and poured onto a microfluidic master containing channel shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. Independently, a second master containing channel-shaped 100- μm structural features spin-coated droplets of a liquid methacrylate unit-terminated PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 2-3 hours under a nitrogen purge. The layer of PDMS was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. A PDMS layer was placed on the surface of the thin layer of partially cured PFPE and heated at 65 ℃ for 10 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a partially cured PFPE smooth layer on a glass slide and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 10

Epoxy/amine

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in stoichiometric ratios and poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The cured layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer was then placed on a clean glass slide and fluid was introduced through the inlet holes.

Example 11

Epoxy/amine excess

Adhered to glass

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in a 4: 1 epoxy to amine ratio such that the epoxy groups were in excess and then poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The cured layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. This layer is then placed on the surface of a clean glass slide, which has been treated with a silane coupling agent, aminopropyltriethoxysilane. The slide was then heated at 65 ℃ for 5 hours to permanently bond the device to the glass slide. Fluid is then introduced through the inlet aperture.

Example 12

Epoxy/amine excess

Bonded to PFPE layer

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in a 1: 4 epoxy to amine ratio such that the amine was in excess and then poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. Independently, a second master containing channel-shaped 100- μm structural features was coated with a liquid PFPE precursor of droplets mixed at a 4: 1 epoxy to amine ratio such that the epoxy units are in excess, and then spin coated at 3700rpm for 1 minute to a thickness of about 20 μm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. A thick layer was placed on the surface of the cured PFPE thin layer and heated at 65 ℃ for 5 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with a silane coupling agent, aminopropyltriethoxysilane, and then heated in an oven at 65 ℃ for 5 hours to bond the device to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 13

Epoxy/amine excess

Adhered to the PDMS layer

A liquid poly (dimethylsiloxane) precursor was poured onto a microfluidic master containing channel-shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. Independently, a second master containing channel-shaped 100- μm structural features was coated with a liquid PFPE precursor of droplets mixed at a 4: 1 epoxy to amine ratio such that the epoxy units are in excess, and then spin coated at 3700rpm for 1 minute to a thickness of about 20 μm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The layer of PDMS was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. The layer was then treated with oxygen plasma for 20 minutes, followed by treatment with a silane coupling agent, aminopropyltriethoxysilane. The treated PDMS layer was placed on the surface of the PFPE thin layer and heated at 65 ℃ for 10 hours to bond the two layers. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with aminopropyltriethoxysilane and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 14

Epoxy/amine excess

Binding to PFPE layer, attachment of biomolecules

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in a 1: 4 epoxy to amine ratio such that the amine excess was mixed together and then poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. Independently, a second master containing channel-shaped 100- μm structural features was coated with a liquid PFPE precursor of droplets mixed at a 4: 1 epoxy to amine ratio such that the epoxy units are in excess, and then spin coated at 3700rpm for 1 minute to a thickness of about 20 μm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. A thick layer was placed on the surface of the cured PFPE thin layer and heated at 65 ℃ for 5 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with a silane coupling agent, aminopropyltriethoxysilane, and then heated in an oven at 65 ℃ for 5 hours to bond the device to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a protein functionalized with free amine is then flowed through the channels lined with unreacted epoxy moieties to functionalize the channels with the protein.

Example 15

Epoxy/amine excess

Bonding to PFPE layers, attachment of charged species

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in a 1: 4 epoxy to amine ratio such that the amine was in excess and then poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. Independently, a second master containing channel-shaped 100- μm structural features was coated with a liquid PFPE precursor of droplets mixed at a 4: 1 epoxy to amine ratio such that the epoxy units are in excess, and then spin coated at 3700rpm for 1 minute to a thickness of about 20 μm. The wafers were then placed in an oven at 65 ℃ for 5 hours. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. A thick layer was placed on the surface of the cured PFPE thin layer and heated at 65 ℃ for 5 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with a silane coupling agent, aminopropyltriethoxysilane, and then heated in an oven at 65 ℃ for 5 hours to bond the device to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a charged molecule functionalized with a free amine is then flowed through a channel lined with unreacted epoxy moieties to functionalize the channel with the charged molecule.

Example 16

Epoxy/amine partial cure

Adhered to glass

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in stoichiometric ratios and poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 0.5 hour, so that it was partially cured. The partially cured layer is then peeled off, trimmed, and an access hole is formed through it using a luer punch. This layer was then placed on a glass slide that had been treated with a silane coupling agent, aminopropyltriethoxysilane, and then heated at 65 ℃ for 5 hours to cause it to adhere to the glass slide. A small needle is then placed into the inlet to introduce the fluid.

Example 17

Epoxy/amine partial cure

Layer-to-layer bonding

Two-component liquid PFPE precursor systems containing PFPE diepoxy and PFPE diamine as shown below were mixed together in stoichiometric ratios and poured onto a microfluidic master containing channel-shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed in an oven at 65 ℃ for 0.5 hour, so that it was partially cured. The partially cured layer is then peeled off, trimmed, and an access hole is formed through it using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed in an oven at 65 ℃ for 0.5 hour, so that it was partially cured. The thick layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The two layers were then placed in an oven and heated at 65 ℃ for 1 hour to bond the two layers. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 18

Epoxy/amine partial cure

PDMS bonding

A liquid poly (dimethylsiloxane) precursor was poured onto a microfluidic master containing channel-shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. The cured PDMS layer was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. The layer was then treated with oxygen plasma for 20 minutes, followed by treatment with a silane coupling agent, aminopropyltriethoxysilane. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with a stoichiometric ratio of small droplets of a liquid PFPE precursor at 3700rpm for 1 minute to a thickness of about 20 μm. The wafers were then placed in an oven at 60 ℃ for 0.5 hour. A treated PDMS layer was placed on the surface of the thin layer of partially cured PFPE and heated at 65 ℃ for 1 hour. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide that had been treated with aminopropyltriethoxysilane and heated at 65 ℃ for 10 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 19

Photocuring with latent functional groups that can be post-cured

Adhered to glass

A liquid PFPE precursor having the structure shown below (where R is an epoxy group, the curve is a PFPE chain, and the ring is a linker molecule) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. The device was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to permanently bond the device to the glass slide. A small needle is then placed into the inlet to introduce the fluid.

Example 20

Photocuring with latent functional groups that can be post-cured

Bonding to PFPE

A liquid PFPE precursor having the structure shown below (where R is an epoxy group, the curve is a PFPE chain, and the ring is a linker molecule) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor (where R is an amine group) at 3700rpm for 1 minute, resulting in a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thick layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 21

Photocuring with latent functional groups that can be post-cured

Adhered to PDMS

A liquid poly (dimethylsiloxane) precursor was poured onto a microfluidic master containing channel-shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. The cured PDMS layer was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. The layer was then treated with oxygen plasma for 20 minutes, followed by treatment with a silane coupling agent, aminopropyltriethoxysilane. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor (where R is an epoxy group) at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. A thicker PDMS layer was then placed on the surface of the 20- μm thick layer and oriented over the desired area to form the seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 22

Photocuring with latent functional groups that can be post-cured

Attachment of biomolecules

A liquid PFPE precursor having the structure shown below (where R is an amine group, the curve is a PFPE chain, and the ring is a linker molecule) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor (where R is an epoxy group) at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a protein functionalized with free amine is then flowed through the channels lined with unreacted epoxy moieties to functionalize the channels with the protein.

Example 23

Photocuring with latent functional groups that can be post-cured

Attachment of charged species

A liquid PFPE precursor having the structure shown below (where R is an amine group, the curve is a PFPE chain, and the ring is a linker molecule) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor (where R is an epoxy group) at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a charged molecule functionalized with a free amine is then flowed through a channel lined with unreacted epoxy moieties to functionalize the channel with the charged molecule.

Example 24

Photocuring with post-curable functional monomers

Adhered to glass

A liquid PFPE dimethacrylate precursor or monomethacrylate PFPE macromonomer was mixed with a monomer having the structure shown below (where R is an epoxy group) and 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. The device was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to permanently bond the device to the glass slide. A small needle is then placed into the inlet to introduce the fluid.

Example 25

Photocuring with post-curable functional monomers

Bonding to PFPE

A liquid PFPE dimethacrylate precursor was mixed with a monomer having the structure shown below (where R is an epoxy group) and 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor plus a functionalized monomer (where R is an amine group) for 1 minute at 3700rpm, resulting in a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 26

Photocuring with post-curable functional monomers

Adhered to PDMS

A liquid poly (dimethylsiloxane) precursor was poured onto a microfluidic master containing channel-shaped 100 μm structural features. The wafers were then placed in an oven at 80 ℃ for 3 hours. The cured PDMS layer was then peeled off, trimmed, and an inlet hole was formed through it using a luer punch. The layer was then treated with oxygen plasma for 20 minutes, followed by treatment with a silane coupling agent, aminopropyltriethoxysilane. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE dimethacrylate precursor plus a functionalized monomer (where R is an epoxy group) plus a photoinitiator for 1 minute at 3700rpm to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. This thicker PDMS layer was then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 27

Photocuring with post-curable functional monomers

Attachment of biomolecules

The liquid PFPE dimethacrylate precursor was mixed with monomers having the structure shown below (where R is an amine group) and 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor plus a functionalized monomer (where R is an epoxy group) for 1 minute at 3700rpm to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a protein functionalized with free amine is then flowed through the channels lined with unreacted epoxy moieties to functionalize the channels with the protein.

Example 28

Photocuring with latent functional groups that can be post-cured

Attachment of charged species

The liquid PFPE dimethacrylate precursor was mixed with monomers having the structure shown below (where R is an amine group) and 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The fully cured layer is then peeled off the master and an access hole is formed by punching using a luer punch. Independently, a second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of a liquid PFPE precursor plus a functionalized monomer (where R is an epoxy group) for 1 minute at 3700rpm to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then placed on the surface of a 20 μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 65 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a glass slide treated with a silane coupling agent, aminopropyltriethoxysilane, and heated at 65 ℃ for 15 hours to allow the device to be permanently bonded to the glass slide. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. An aqueous solution containing a charged molecule functionalized with a free amine is then flowed through a channel lined with unreacted epoxy moieties to functionalize the channel with the charged molecule.

Example 29

Fabrication of PFPE microfluidic devices using sacrificial channels

A smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE dimethacrylate precursor onto a glass slide with a spatula. The slides were then placed in a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. A scaffold (scaffold) consisting of a channel-shaped poly (lactic acid) was arranged on a flat, smooth layer of PFPE. The liquid PFPE dimethacrylate precursor was mixed with 1 wt% of a free radical photoinitiator and poured onto the scaffold. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The device was then placed in a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The device was then heated at 150 ℃ for 24 hours to degrade the poly (lactic acid), thus exposing the remaining voids in the shape of channels.

Example 30

Bonding PFPE devices to glass using 185nm light

The liquid PFPE dimethacrylate precursor was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing channel shaped 100 μm structural features. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer was then placed on the surface of a 20 μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 120 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded two layers were then placed on a clean glass slide to form a seal. The device was exposed to 185nm UV light for 20 minutes to form a permanent bond between the device and the glass. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 31

Epoxy sleeve sealing method for packaging equipment

The liquid PFPE dimethacrylate precursor was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing channel shaped 100 μm structural features. The area required for the PDMS mold contained liquid to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 120 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layers were then placed on a clean glass slide, requiring the formation of a seal. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6. The entire apparatus is then encased in a liquid epoxy precursor, which is poured over the equipment and allowed to cure. The sleeve is used to mechanically bond the device and the substrate.

Example 32

Manufacture of PFPE equipment from three-arm (three-arm) PFPE precursors

A liquid PFPE precursor having the structure shown below (where the ring is a linker molecule) was mixed with 1 wt% of a free radical photoinitiator and poured onto a microfluidic master containing 100 μm structural features of channel shape. A PDMS mold was used to contain the liquid in the desired area to a thickness of about 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Independently, a second master containing channel-shaped 100- μm structural features was spin coated with droplets of the liquid PFPE precursor on its surface at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. Third, a smooth, flat layer of PFPE was formed by scraping a small droplet of the liquid PFPE precursor onto a glass slide with a spatula. The slides were then placed in a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The thicker layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The layer is then placed on the surface of a 20- μm thick layer and oriented over the desired area to form a seal. The layer was then placed in an oven and heated at 120 ℃ for 2 hours. The thin layer is then trimmed and the adhered layers are lifted from the master. The fluid inlet and outlet holes are die cut by using a luer punch. The bonded layer was then placed on a fully cured PFPE smooth layer on a glass slide and heated at 120 ℃ for 15 hours. A small needle is then placed into the inlet to introduce fluid and activate the membrane valve as reported by Unger, m. et al, science.2000, 288, 113-6.

Example 33

Photocuring PFPE/PDMS hybridization

A second master containing channel-shaped 100- μm structural features was spin coated on its surface with droplets of the liquid PFPE dimethacrylate precursor at 3700rpm for 1 minute to a thickness of about 20 μm. PDMS dimethacrylate containing a photoinitiator was then poured onto the surface of the thin PFPE layer to a thickness of 3 mm. The wafer was then placed into a UV chamber and exposed to UV light (λ -365) for 10 minutes under a nitrogen purge. The layer is then peeled off, trimmed, and an entry hole is formed through it using a luer punch. The hybrid device is then placed on a glass slide and a seal is formed. A small needle is then placed into the inlet to introduce the fluid.

It will be understood that various details of the presently disclosed subject matter can be changed without departing from the scope of the presently disclosed subject matter. Furthermore, the foregoing description is for the purpose of illustration only, and not for the purpose of limitation.

Claims

1. A microfluidic device comprising a perfluoropolyether (PFPE) material, wherein the PFPE material is prepared from a liquid PFPE material having properties selected from the group consisting of: (i) a viscosity of greater than about 100 centistokes (cSt), (ii) a viscosity of less than about 100cSt, with the proviso that the liquid PFPE precursor material having a viscosity of less than 100cSt is not a free radical photo-curable PFPE material, and (iii) combinations thereof.

2. The microfluidic device of claim 1, wherein the liquid PFPE precursor is terminated with a polymerizable group.

3. The microfluidic device of claim 2, wherein the polymerizable group is selected from the group consisting of acrylate, methacrylate, epoxy, amino, carboxyl, anhydride, maleimide, isocyanate, olefin, and styrene groups.

4. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises a backbone structure, wherein the backbone structure is selected from the group consisting of:

wherein:

x is present or absent and when present includes a capping group, and

n is an integer from 1 to 100.

5. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises the following structure:

Wherein n is an integer from 1 to 100.

6. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises the following structure:

wherein n is an integer from 1 to 100.

7. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises a compound comprising the structure:

wherein: wherein the ring comprises a multifunctional linker molecule; and the PFPE includes a perfluoropolyether chain.

8. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises a highly branched PFPE liquid precursor material.

9. The microfluidic device of claim 1, wherein the liquid PFPE material comprises an end-functionalized material selected from the group consisting of:

10. the microfluidic device of claim 1, wherein the liquid PFPE material comprises a functionalized monomer.

11. The microfluidic device according to claim 10, wherein the functionalized monomer is selected from the group consisting of styrenes, methacrylates, acrylates, acrylamides, acrylonitriles, and vinylpyridines.

12. The microfluidic device of claim 11, wherein the styrenic is selected from the group consisting of pentafluorostyrene, bromostyrene, chlorostyrene, styrene sulfonic acid, fluorostyrene, and styrene acetate.

13. The microfluidic device of claim 11, wherein the methacrylate is selected from the group consisting of t-butyl methacrylate, dimethylaminopropyl methacrylate, glycidyl methacrylate, hydroxyethyl methacrylate, aminopropyl methacrylate, cyanomethacrylate, trimethoxysilane methacrylate, isocyanato methacrylate, lactone-containing methacrylate, saccharide-containing methacrylate, polyethylene glycol methacrylate, norbornane-containing methacrylate, polyhedral oligomeric silsesquioxane methacrylate, 2-trimethylsiloxyethyl methacrylate, and 1H, 2H-fluorooctyl methacrylate.

14. The microfluidic device of claim 11, wherein the acrylate is selected from the group consisting of t-butyl acrylate, allyl acrylate, cyanoacrylate, trimethoxysilane acrylate, lactone-containing acrylate, sugar-containing acrylate, polyethylene glycol acrylate, and norbornane-containing acrylate.

15. The microfluidic device of claim 1, wherein the liquid PFPE precursor material comprises a two-component liquid PFPE precursor system comprising a mixture of two functionalized PFPE components mixed in a stoichiometric ratio.

16. The microfluidic device of claim 15, wherein the two-component PFPE precursor system comprises a mixture of components selected from the group consisting of epoxy/amine mixtures, hydroxyl/isocyanate mixtures, hydroxyl/acid chloride mixtures, and hydroxyl/chlorosilane mixtures.

17. The microfluidic device of claim 16, wherein the epoxy/amine mixture comprises a PFPE diepoxy compound comprising the structure:

and, a PFPE diamine compound comprising the structure:

18. the microfluidic device of claim 16, wherein the epoxy/amine mixture comprises a stoichiometric ratio in a range of about 4: 1 epoxy to amine to about 1: 4 epoxy to amine.

19. The microfluidic device of claim 1, wherein the liquid PFPE precursor material is mixed with a functionalizing substance, wherein the functionalizing substance mechanically entangles into a PFPE network after curing.

20. The microfluidic device of claim 1, wherein the perfluoropolyether (PFPE) material comprises a thermally cured liquid PFPE precursor material.

21. The microfluidic device of claim 1, wherein the perfluoropolyether (PFPE) material comprises a chemically cured liquid PFPE precursor material.

22. The microfluidic device of claim 1, wherein the perfluoropolyether (PFPE) material comprises a photoacid-cured liquid PFPE precursor material.

23. The microfluidic device of claim 1, wherein the PFPE material is transparent to one of UV light, visible light, and combinations thereof.

24. A microfluidic device comprising a fluoroolefin-based elastomer, wherein the fluoroolefin-based elastomer comprises a first monomer and at least one additional monomer, wherein the first monomer and the at least one additional monomer are different, wherein:

(a) the first monomer is selected from vinylidene fluoride and tetrafluoroethylene; and

(b) the at least one additional monomer is selected from the group consisting of fluorine-containing olefins, fluorine-containing vinyl ethers, olefins, and combinations thereof.

25. The microfluidic device of claim 24, wherein the fluorine-containing olefin is selected from the group consisting of vinylidene fluoride, Hexafluoropropylene (HFP), Tetrafluoroethylene (TFE), 1, 2, 3, 3, 3-pentafluoropropene (1-HPFP), Chlorotrifluoroethylene (CTFE), and vinyl fluoride.

26. The microfluidic device of claim 24, wherein the fluorine-containing vinyl ether comprises a perfluoro (alkyl vinyl) ether.

27. The microfluidic device according to claim 24, wherein the olefin is selected from ethylene and propylene.

28. The microfluidic device of claim 24, wherein the fluoroolefin-based elastomer comprises copolymerized units of:

vinylidene fluoride and hexafluoropropylene;

vinylidene fluoride, hexafluoropropylene, and tetrafluoroethylene;

vinylidene fluoride, hexafluoropropylene, tetrafluoroethylene, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1;

vinylidene fluoride, hexafluoropropylene, tetrafluoroethylene, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1;

vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1;

vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1;

vinylidene fluoride, perfluoro (methyl vinyl) ether, tetrafluoroethylene, and 1, 1, 3, 3, 3-pentafluoropropene;

tetrafluoroethylene, perfluoro (methyl vinyl) ether, and ethylene;

tetrafluoroethylene, perfluoro (methyl vinyl) ether, ethylene, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1;

tetrafluoroethylene, perfluoro (methyl vinyl) ether, ethylene, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1;

tetrafluoroethylene, propylene, and vinylidene fluoride;

tetrafluoroethylene and perfluoro (methyl vinyl) ether;

tetrafluoroethylene, perfluoro (methyl vinyl) ether, and perfluoro (8-cyano-5-methyl-3, 6-dioxa-1-octene);

Tetrafluoroethylene, perfluoro (methyl vinyl) ether, and 4-bromo-3, 3, 4, 4-tetrafluorobutene-1;

tetrafluoroethylene, perfluoro (methyl vinyl) ether, and 4-iodo-3, 3, 4, 4-tetrafluorobutene-1; and

tetrafluoroethylene, perfluoro (methyl vinyl) ether, and perfluoro (2-phenoxypropyl vinyl) ether.

29. The microfluidic device of claim 24, wherein the fluoroolefin-based elastomer comprises at least one cure site monomer.

30. The microfluidic device according to claim 29, wherein the cure site monomer is selected from the group consisting of a bromine-containing olefin, an iodine-containing olefin, a bromine-containing vinyl ether, an iodine-containing vinyl ether, a fluorine-containing olefin comprising a nitrile group, a fluorine-containing vinyl ether comprising a nitrile group, 1, 3, 3, 3-pentafluoropropene (2-HPFP), perfluoro (2-phenoxypropyl vinyl) ether, and a non-conjugated diene.

31. The microfluidic device of claim 24, wherein the fluoroolefin-based elastomer is transparent to one of UV light, visible light, and combinations thereof.

32. The microfluidic device of claim 24, wherein the fluoroolefin-based elastomer has a mooney viscosity of less than about 40(ML1+10 at 121 ℃).

33. The microfluidic device of claim 24, wherein the fluoroolefin-based elastomer is permeable to oxygen, carbon dioxide, and nitrogen.

34. A method of surface functionalization of a microdevice, the method comprising forming a layer of a functionalizing material, wherein the functionalizing material is selected from a liquid PFPE precursor material and a liquid fluoroolefin-type precursor material.

35. The method of claim 34, wherein the layer of functionalized material includes latent functional groups that do not react during curing.

36. The method of claim 35, wherein the latent functional group comprises a methacrylate group.

37. The method of claim 34, wherein the layer of functionalizing material comprises latent functional groups that are introduced in the generation of a liquid precursor material.

38. The method of claim 37, wherein the latent functional group comprises a methacrylate group.

39. The method of claim 34, wherein the layer of functionalized material comprises a two-component liquid PFPE precursor material, wherein the two-component liquid PFPE precursor material comprises a mixture of two functionalized PFPE components mixed in a stoichiometric ratio.

40. The method of claim 34, wherein the layer of functionalized material comprises chemical linker groups.

41. The method of claim 40, wherein the chemical linker group comprises the following structure:

Wherein:

r comprises an epoxy group;

wherein the circular ring comprises a linker molecule; and

the wavy line in the formula includes a PFPE chain.

42. The method of claim 34, wherein the layer of functionalized material comprises a functionalized monomer.

43. The method of claim 42, wherein the functionalized monomer is selected from the group consisting of t-butyl methacrylate, t-butyl acrylate, dimethylaminopropyl methacrylate, glycidyl methacrylate, hydroxyethyl methacrylate, aminopropyl methacrylate, allyl acrylate, cyanoacrylate, cyanomethacrylate, trimethoxy silane acrylate, trimethoxy silane methacrylate, isocyanato methacrylate, lactone-containing acrylate, lactone-containing methacrylate, sugar-containing acrylate, sugar-containing methacrylate, polyethylene glycol methacrylate, norbornane-containing acrylate, polyhedral oligomeric silsesquioxane methacrylate, 2-trimethylsiloxyethyl methacrylate, poly (ethylene glycol) methacrylate, poly (, 1H, 1H, 2H, 2H-fluorooctyl methacrylate, pentafluorostyrene, vinylpyridine, bromostyrene, chlorostyrene, styrenesulfonic acid, fluorostyrene, styrene acetate, acrylamide, and acrylonitrile.

44. The method of claim 34, wherein the layer of functionalizing material is functionalized by exposure to a plasma.

45. The method of claim 44, wherein the plasma is selected from the group consisting of an argon plasma and an oxygen plasma.

46. The method of claim 34, wherein the layer of functionalizing material is functionalized by exposure to ultraviolet radiation.

47. The method of claim 34, comprising attaching a functionalized moiety to the layer of functionalized material.

48. The method of claim 47, wherein the functional moiety is selected from the group consisting of a protein, an oligonucleotide, a drug, a catalyst, a dye, a sensor, an analyte, and a charged species capable of altering the wettability of a channel.

49. The method of claim 34, wherein the layer of functionalized material comprises a microfluidic channel.

50. The method of claim 34, comprising bonding the layer of functionalized material to a substrate.

51. The method of claim 50, wherein the substrate comprises a microtitre tube.

52. A layer of a functionalized material prepared by the method of claim 34.

53. A method of forming a multi-layer device, the method comprising:

(a) Providing a first layer of material, wherein the first layer of material comprises a material selected from the group consisting of: a liquid perfluoropolyether (PFPE) precursor, a poly (dimethylsiloxane) (PDMS) precursor, a polyurethane precursor comprising PDMS blocks, a precursor comprising PFPE and PDMS blocks, and a fluoroolefin-type precursor; and

(b) contacting the first layer of material with:

(i) a substrate;

(ii) a second layer of material, wherein the second layer of material comprises a material selected from the group consisting of: a perfluoropolyether (PFPE) precursor, a poly (dimethylsiloxane) (PDMS) precursor, a polyurethane precursor comprising PDMS blocks, a precursor comprising PFPE and PDMS blocks, and a fluoroolefin-type precursor; and wherein the second layer of material may be the same or different from the first layer of material; and

(iii) combinations thereof; to form a multi-layer device.

54. The method of claim 53, wherein the first layer of material comprises a fully cured material.

55. The method of claim 53, wherein the first layer of material forms a reversible seal in contact with the substrate.

56. The method of claim 53, wherein the first layer of material comprises a partially cured material.

57. The method of claim 56, wherein the partially cured material comprises a partially cured PFPE precursor material terminated with methacrylate groups.

58. The method of claim 53, comprising treating the substrate with a silane coupling agent to form a treated substrate.

59. The method of claim 58, wherein the silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein said monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes are functionalized with a moiety selected from the group consisting of: amines, methacrylates, acrylates, styrenes, epoxies, isocyanates, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

60. The method of claim 56, comprising:

(a) contacting the first layer of partially cured material with the treated substrate; and

(b) the first layer of partially cured material is treated to form a bond between the first layer of partially cured material and the treated substrate.

61. The method of claim 53, wherein:

(a) The first layer of material comprises a first partially cured material; and

(b) the second layer of material comprises a second partially cured material, wherein the first partially cured material and the second partially cured material may be the same or different.

62. The method of claim 61, comprising:

(a) contacting a first layer of partially cured material with a second layer of partially cured material to form a partially cured multilayer device; and

(b) processing the partially cured multilayer device to form a fully cured multilayer device.

63. The method of claim 62, wherein the processing comprises a process selected from the group consisting of: thermal curing processes, chemical curing processes, photoacid generator curing processes, and catalytic curing processes.

64. The method of claim 62, wherein the first layer of partially-cured material and the second layer of partially-cured material each comprise a thermally-curable PFPE precursor material.

65. The method of claim 62, wherein the first layer of partially cured material comprises a polyurethane precursor material and the second layer of partially cured material comprises a PFPE precursor material.

66. The method of claim 62, wherein the first layer of partially cured material comprises a polyurethane precursor comprising poly (dimethylsiloxane) blocks and the second layer of partially cured material comprises a PFPE precursor material.

67. The method of claim 62, wherein the first layer of partially cured material comprises a precursor material comprising PFPE blocks and PDMS blocks, and the second layer of partially cured material comprises a PFPE precursor material.

68. The method of claim 62, wherein the first layer of partially cured material comprises a PDMS precursor and the second layer of partially cured material comprises a PFPE precursor material.

69. The method of claim 68, wherein the PFPE precursor material is end-capped with a methacrylate group.

70. The method of claim 68, comprising treating the PDMS precursor with a plasma treatment followed by treatment with a silane coupling agent.

71. The method as claimed in claim 70, wherein the silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein said monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes are functionalized with a moiety selected from the group consisting of: amines, methacrylates, acrylates, styrenes, epoxies, isocyanates, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

72. The method of claim 62, comprising:

(a) contacting the partially cured multilayer structure with a substrate, wherein the substrate is coated with a partially cured precursor material to form a second partially cured multilayer device; and

(b) processing the second partially cured multilayer device to form a second fully cured multilayer device.

73. The method of claim 72, wherein the processing comprises a process selected from: thermal curing processes, chemical curing processes, photoacid generator curing processes, and catalytic curing processes.

74. The method of claim 53, wherein at least one of the first layer of material and the second layer of material comprises a material formed from a two-component PFPE precursor material, wherein the two-component PFPE precursor material comprises a mixture of two functionalized PFPE components mixed in a stoichiometric ratio.

75. The method of claim 74, wherein the two-component PFPE precursor system comprises a mixture of components selected from the group consisting of: epoxy/amine mixtures, hydroxyl/isocyanate mixtures, hydroxyl/acid chloride mixtures and hydroxyl/chlorosilane mixtures.

76. The process of claim 75 wherein said epoxy/amine mixture comprises a PFPE diepoxy compound comprising the structure:

And, a PFPE diamine compound comprising the structure:

77. the method of claim 75 wherein the epoxy/amine mixture comprises a stoichiometric ratio within the range of about 4: 1 epoxy to amine to about 1: 4 epoxy to amine.

78. The method of claim 77, wherein the stoichiometric ratio is about 4: 1 epoxyamine.

79. The method of claim 78, comprising:

(a) providing a substrate, wherein the substrate is treated with a silane coupling agent;

(b) contacting a first layer of a material formed from a two-component PFPE precursor material comprising a stoichiometric ratio of about 4: 1 epoxy to amine with a substrate; and

(b) the first layer of material and the substrate are processed to form a multi-layer device.

80. The method of claim 79, wherein the silane coupling agent comprises aminopropyltriethoxysilane.

81. The method of claim 77, wherein the stoichiometric ratio is about 1: 4 epoxy to amine.

82. The method of claim 81, comprising:

(i) providing a first layer of material comprising a stoichiometric ratio of about 1: 4 epoxy to amine;

(ii) contacting a first layer comprising about a 1: 4 stoichiometric ratio of epoxy to amine materials with a second layer comprising about a 4: 1 stoichiometric ratio of epoxy to amine materials; and

(iii) Two layers of the material are processed to form a multi-layer device.

83. The method of claim 78, comprising:

(i) providing a first layer of PDMS material;

(ii) treating the first layer of PDMS material with a plasma treatment process followed by treatment with a silane coupling agent to form a treated layer of PDMS material;

(iii) contacting the handle layer of PDMS material with a second layer of material comprising a stoichiometric ratio of epoxy to amine of about 4: 1; and

(iv) two layers of the material are processed to form a multi-layer device.

84. The method of claim 83, wherein the silane coupling agent comprises aminopropyltriethoxysilane.

85. The method of claim 74, comprising:

(a) providing a first layer of material formed from a two-component PFPE precursor material, wherein the two-component PFPE precursor material comprises a mixture of two functionalized PFPE components mixed in a stoichiometric ratio;

(b) treating the first layer of material to form a first layer of partially cured material;

(c) contacting the first layer of partially cured material with one of:

(i) a substrate;

(ii) a second layer of material; and

(iii) combinations thereof; and

(d) treating the first layer of partially cured material to bond the partially cured material to one of a substrate, a second layer of material, and combinations thereof.

86. The method of claim 85, wherein the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, and a fused silica material.

87. The method of claim 86, comprising treating the substrate with a silane coupling agent.

88. The method as claimed in claim 87, wherein the silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein said monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes are functionalized with a moiety selected from the group consisting of: amines, methacrylates, acrylates, styrenes, epoxies, isocyanates, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

89. The method of claim 85, wherein the second layer of material comprises a PFPE precursor material.

90. The method of claim 85, wherein the second layer of material comprises a poly (dimethylsiloxane) material, wherein the poly (dimethylsiloxane) material is treated with oxygen plasma followed by treatment with a silane coupling agent.

91. The method of claim 53, wherein the PFPE precursor material comprises the structure:

wherein:

r comprises an epoxy group;

wherein the circular ring comprises a linker molecule; and

the wavy line in the formula includes a PFPE chain.

92. The method of claim 91, comprising photocuring the PFPE precursor material to form a layer of fully cured PFPE material.

93. The method of claim 92, comprising:

(a) contacting the layer of fully cured PFPE material with one of:

(i) a substrate;

(ii) a second layer of material; and

(iii) combinations thereof; and

(b) treating the fully cured material to bond with one of the substrate, the second layer of material, and combinations thereof.

94. The method of claim 93, wherein the substrate is selected from a glass material, a quartz material, a silicon material, and a fused silica material.

95. The method of claim 94, comprising treating the substrate with a silane coupling agent.

96. The method of claim 95, wherein the silane coupling agent comprises aminopropyltriethoxysilane.

97. The method of claim 93, wherein the second layer of material comprises a PFPE material.

98. The method of claim 93, wherein the second layer of material comprises a treated PDMS material, wherein the treated PDMS material is treated with an oxygen plasma followed by treatment with a silane coupling agent.

99. The method of claim 98, wherein the silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein said monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes are functionalized with a moiety selected from the group consisting of: amines, methacrylates, acrylates, styrenes, epoxies, isocyanates, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

100. The method of claim 53, comprising mixing a PFPE precursor with a functionalizing monomer to form a PFPE precursor mixture.

101. The method of claim 100, wherein the functionalized monomer comprises the following structure:

102. the method of claim 100, comprising photocuring the PFPE precursor mixture to form a layer of fully cured PFPE material.

103. The method of claim 102, comprising:

(a) contacting the layer of fully cured PFPE material with one of:

(i) a substrate;

(ii) a second layer of material; and

(iii) combinations thereof; and

(b) treating the layer of fully cured material to bond with one of a substrate, a second layer of material, and combinations thereof.

104. The method of claim 103, wherein the substrate is selected from a glass material, a quartz material, a silicon material, and a fused silica material.

105. The method of claim 104, comprising treating the substrate with a silane coupling agent.

106. The process as set forth in claim 105 wherein said silane coupling agent is selected from the group consisting of monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes; and wherein said monohalosilanes, dihalosilanes, trihalosilanes, monoalkoxysilanes, dialkoxysilanes, and trialkoxysilanes are functionalized with a moiety selected from the group consisting of: amines, methacrylates, acrylates, styrenes, epoxies, isocyanates, halogens, alcohols, benzophenone derivatives, maleimides, carboxylic acids, esters, acid chlorides, and olefins.

107. The method of claim 103, wherein the second layer of material comprises PFPE material.

108. The method of claim 103, wherein the second layer of material comprises a treated PDMS material, wherein the treated PDMS material is treated with an oxygen plasma followed by treatment with a silane coupling agent.

109. The method of claim 108, wherein the silane coupling agent comprises aminopropyltriethoxysilane.

110. The method of claim 53, wherein the substrate is selected from a glass material, a quartz material, a silicon material, a fused silica material, an elastomeric material, and a rigid thermoplastic material.

111. The method of claim 110, wherein the elastomeric material is selected from the group consisting of poly (dimethylsiloxane) (PDMS), Kratons, buna rubber, natural rubber, fluoroelastomers, chloroprene, butyl rubber, nitrile rubber, polyurethane, and thermoplastic elastomers.

112. The method of claim 110, wherein the rigid thermoplastic material is selected from the group consisting of polystyrene, poly (methyl methacrylate), polyester, polycarbonate, polyimide, polyamide, polyvinyl chloride, polyolefin, poly (ketone), poly (ether ketone), and poly (ether sulfone).

113. The method of claim 110, comprising treating the substrate with a silane coupling agent.

114. A process as set forth in claim 113 wherein said silane coupling agent is selected from the group consisting of trimethylsilylpropyl methacrylate and aminopropyltriethoxysilane.

115. The method of claim 53, wherein the substrate comprises a microtiter plate.

116. The method of claim 53, wherein the first layer of material comprises at least one microscale channel.

117. The method of claim 53, wherein the first layer of material comprises at least one nanoscale channel.

118. A multi-layer device formed by the method of claim 53.

119. The multi-layer device of claim 118, wherein the multi-layer device comprises a microfluidic device.

120. A method of bonding one of a microscale device, a nanoscale device, and combinations thereof, to a substrate, the method comprising:

(a) providing one of a micro-scale device, a nano-scale device, and combinations thereof, wherein the device comprises a material selected from the group consisting of: perfluoropolyether materials and fluoroalkene-type materials;

(b) contacting the device with a substrate;

(c) Coating the device and the substrate with a liquid precursor encapsulation material;

(d) solidifying the liquid precursor encapsulation material, thereby mechanically bonding the device to the substrate.

121. The method of claim 120, wherein the substrate is selected from the group consisting of a glass material, a quartz material, a silicon material, a fused silica material, an elastomeric material, and a rigid thermoplastic material.

122. The method of claim 121, wherein the elastomeric material is selected from the group consisting of poly (dimethylsiloxane) (PDMS), Kratons, buna rubber, natural rubber, fluoroelastomers, chloroprene, butyl rubber, nitrile rubber, polyurethane, and thermoplastic elastomers.

123. The method of claim 121, wherein the rigid thermoplastic material is selected from the group consisting of polystyrene, poly (methyl methacrylate), polyester, polycarbonate, polyimide, polyamide, polyvinyl chloride, polyolefin, poly (ketone), poly (ether ketone), and poly (ether sulfone).

124. The method of claim 120, wherein the substrate is treated with a silane coupling agent.

125. A process as set forth in claim 124 wherein said silane coupling agent is selected from the group consisting of trimethylsilylpropyl methacrylate and aminopropyltriethoxysilane.

126. The method of claim 120, wherein solidifying the liquid precursor encapsulation material comprises a curing process.

127. The method of claim 120, wherein the liquid precursor encapsulating material is selected from a liquid epoxy-based precursor and a polyurethane.

128. A method of forming one of micro-scale structures, nano-structures, and combinations thereof, the method comprising:

(a) disposing a first PFPE precursor material on a substrate, thereby forming a first layer of liquid PFPE precursor material on the substrate;

(b) treating the first layer of PFPE precursor material to form a first layer of treated PPFE material on the substrate;

(c) placing a multi-dimensional structure on a first layer of processed PFPE material, wherein the multi-dimensional structure has a property selected from the group consisting of: (i) degradability; (ii) is selectively soluble; and (iii) combinations thereof;

(d) encapsulating the multi-dimensional structure with a second layer of liquid PFPE precursor material;

(e) treating the second layer of PFPE precursor material, thereby forming a second layer of treated PFPE material; and

(f) removing the degradable or selectively soluble material from the second layer of treated PFPE material to form one of a micro-scale structure, a nano-scale structure, and combinations thereof.

129. The method of claim 128, wherein the degradable or selectively soluble material is selected from the group consisting of waxes, photoresists, poly (lactic acid), polylactones, polysulfones, polyelectrolytes, cellulosic fibers, water soluble polymers, solvent soluble polymers, salts, solid organic compounds, and solid inorganic compounds.

130. The method of claim 128, wherein the removing of the degradable or selectively soluble material comprises a process selected from the group consisting of: thermal processes, photochemical processes and dissolution processes.

131. The method of claim 128, comprising mixing at least one of a first PFPE precursor material and a second PFPE precursor material with one of a thermal radical initiator and a photoinitiator.

132. The method of claim 128, wherein the processing of at least one of the first layer of PFPE precursor material and the second layer of PFPE precursor material comprises a curing process.

133. The method of claim 132, wherein the curing process is selected from the group consisting of a thermal curing process and a photochemical curing process.

134. The method of claim 128, wherein encapsulating the multi-dimensional structure with a second layer of liquid PFPE precursor material comprises a spin-on process.

135. A micron-scale structure made by the method of claim 128.

136. The micro-scale structure of claim 135, wherein the micro-scale structure comprises a microfluidic channel.

137. A nanostructure prepared by the method of claim 128.

138. The nanostructure of claim 137, wherein the nanostructure comprises a nanoscale channel.

139. A method of forming one of micro-scale structures, nano-structures, and combinations thereof, the method comprising:

(a) providing a patterned layer of perfluorinated perfluoropolyether (PFPE) material, wherein the patterned layer of PFPE material comprises a patterned surface;

(b) disposing a predetermined volume of a degradable or selectively soluble material on the patterned surface of the patterned layer of PFPE material;

(c) encapsulating the predetermined volume of degradable or selectively soluble material on the patterned surface of the patterned layer of PFPE material; and

(d) removing the predetermined volume of degradable or selectively soluble material from the patterned surface of the layer of PFPE material to form one of a micro-scale structure, a nano-scale structure, and combinations thereof.

140. The method of claim 139, wherein the degradable or selectively soluble material is selected from the group consisting of waxes, photoresists, poly (lactic acid), polylactones, polysulfones, polyelectrolytes, cellulosic fibers, water soluble polymers, solvent soluble polymers, salts, solid organic compounds, and solid inorganic compounds.

141. The method of claim 140 wherein the removing of the predetermined volume of degradable or selectively soluble material comprises a process selected from the group consisting of: thermal processes, photochemical processes and dissolution processes.

142. A micron-scale structure made by the method of claim 139.

143. The micro-scale structure of claim 142, wherein the micro-scale structure comprises a microfluidic channel.

144. A nanostructure prepared by the method of claim 139.

145. The nanostructure of claim 144, wherein the nanostructure comprises a nanoscale channel.

146. A method of flowing a material in a microfluidic device, the method comprising:

(a) providing a microfluidic device comprising at least one layer selected from the group consisting of:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof; and

(b) the material is flowed in the micron-scale channels.

147. The method of claim 146, wherein the at least one layer of material covers a surface of at least one of the one or more micro-scale channels.

148. The method of claim 147, wherein at least one layer of the material comprises a functionalized surface.

149. The method of claim 146, wherein the one or more microscale channels comprise an integrated network of microscale channels.

150. The method of claim 149, wherein the microscale channels of the integrated network intersect at a predetermined point.

151. The method of claim 146, wherein the microfluidic device comprises one or more patterned layers of a first polymeric material, and wherein the one or more patterned layers of the first polymeric material define one or more microscale channels.

152. The method of claim 151, wherein the microfluidic device further comprises a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of the first polymeric material.

153. The method of claim 151, wherein at least one patterned layer of the material comprises a functionalized surface.

154. The method of claim 151, wherein the one or more microscale channels comprise an integrated network of microscale channels.

155. The method of claim 154, wherein the microscale channels of the integrated network intersect at a predetermined point.

156. The method of claim 151, wherein the patterned layer of the first polymeric material comprises a plurality of holes.

157. The method of claim 156, wherein at least one of the plurality of apertures includes an inlet aperture.

158. The method of claim 156, wherein at least one of the plurality of apertures comprises an exit aperture.

159. The method of claim 156, wherein the microfluidic device comprises one or more valves.

160. The method of claim 146, wherein the material is selected from the group consisting of a fluid, an organic solvent, an aqueous solution dispersed in a substantially non-aqueous solvent, a surfactant mixture, and a reaction mixture.

161. The method of claim 146, wherein the material flows in a predetermined direction along the microscale channel.

162. The method of claim 146, comprising applying a driving force to move the material along the micro-scale channels.

163. A method of mixing two or more materials, the method comprising:

(a) providing a micro-scale device comprising at least one layer selected from the group consisting of:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof; and

(b) the first material and the second material are contacted in the apparatus to mix the first and second materials.

164. The method of claim 163, wherein the micro-scale device is selected from a microfluidic device and a microtiter plate.

165. The method of claim 164, wherein the microfluidic device comprises one or more microscale channels.

166. The method of claim 165, wherein at least one layer of the material covers a surface of at least one of the one or more micro-scale channels.

167. The method of claim 166, wherein at least one layer of the material comprises a functionalized surface.

168. The method of claim 165, wherein the microfluidic device comprises at least one patterned layer of a first polymeric material, and wherein the patterned layer of the first polymeric material defines one or more microscale channels.

169. The method of claim 168, wherein the microfluidic device further comprises a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of the first polymeric material.

170. The method of claim 168, wherein the patterned layer of the first polymeric material comprises a functionalized surface.

171. The method of claim 165, wherein the one or more microscale channels comprise an integrated network of microscale channels.

172. The method of claim 171, wherein the microscale channels of the integrated network intersect at predetermined points.

173. The method of claim 165, wherein contacting the first material and the second material is performed in a mixing zone defined in the one or more microscale channels.

174. The method of claim 173, wherein the mixing zone comprises a geometry selected from the group consisting of: t-connections, coils, long channels, micron chambers, and constrictions.

175. The method of claim 165, wherein the first material and the second material are disposed in separate channels of a microfluidic device.

176. The method of claim 175, wherein contacting the first material and the second material is performed in a mixing zone defined by intersections of the channels.

177. The method of claim 176, wherein the mixing zone comprises a geometry selected from the group consisting of: t-connections, coils, long channels, micron chambers, and constrictions.

178. The method of claim 164, comprising flowing the first material and the second material in a predetermined direction in a microfluidic device.

179. The method of claim 164, comprising flowing the mixed material in a predetermined direction in a microfluidic device.

180. The method of claim 164, comprising contacting the mixed material with a third material to form a second mixed material.

181. The method of claim 164, comprising flowing the mixed material to an outlet port of a microfluidic device.

182. The method of claim 164, comprising applying a driving force to cause the material to move through the microfluidic device.

183. The method of claim 164, wherein the microtiter plate comprises one or more tubes.

184. The method of claim 183, wherein the at least one layer of material covers a surface of at least one of the one or more tubes.

185. The method of claim 184, wherein at least one layer of the material comprises a functionalized surface.

186. The method of claim 163, comprising recovering the mixed material.

187. A method of screening a sample for a property, the method comprising:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof;

(b) providing a target material;

(c) placing the sample in a micron-scale apparatus;

(d) contacting the sample with the target material; and

(e) detecting an interaction between the sample and the target material,

wherein the presence or absence of the interaction is indicative of a characteristic of the sample.

188. The method of claim 187, wherein said micro-scale device is selected from the group consisting of a microfluidic device and a microtiter plate.

189. The method of claim 188, wherein the microfluidic device comprises one or more microscale channels.

190. The method of claim 189, wherein at least one layer of the material covers a surface of at least one of the one or more microscale channels.

191. The method of claim 189, wherein the microfluidic device comprises at least one patterned layer of a first polymeric material, and wherein the patterned layer of the first polymeric material defines the one or more microscale channels.

192. The method of claim 191, wherein the microfluidic device further comprises a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of the first polymeric material.

193. The method of claim 191, wherein the one or more microscale channels comprise an integrated network of microscale channels.

194. The method of claim 193, wherein microscale channels of the integrated network intersect at predetermined points.

195. The method of claim 188, wherein the microtiter plate comprises one or more tubes.

196. The method of claim 195, wherein the at least one layer of material covers a surface of at least one of the one or more tubes.

197. The method of claim 187, comprising placing a target material in a micro-scale device.

198. The method of claim 197, wherein the target material is bound to a functionalized surface.

199. The method of claim 187, wherein the target material comprises one or more of an antigen, an antibody, an enzyme, a restriction enzyme, a dye, a fluorescent dye, a sequence reagent, a PCR reagent, a primer, a receptor, a ligand, a chemical reagent, or a combination thereof.

200. The method of claim 187, wherein a sample is bound to the functionalized surface.

201. The method of claim 187 wherein the sample is selected from the group consisting of therapeutic agents, diagnostic agents, research reagents, catalysts, metal ligands, non-biological organic materials, inorganic materials, food products, soil, water and air.

202. The method of claim 187 wherein the sample comprises one or more constituent members of one or more libraries of chemical or biological compounds or components.

203. The method of claim 187, wherein the sample comprises one or more of nucleic acid templates, sequence reagents, primers, primer extension products, restriction enzymes, PCR reagents, PCR reaction products, or combinations thereof.

204. The method of claim 187 wherein the sample comprises one or more of an antibody, a cellular receptor, an antigen, a receptor ligand, an enzyme, a substrate for an enzyme, an immunochemical, an immunoglobulin, a virus, a viral binding component, a protein, a cytokine, a growth factor, an inhibitor or a combination thereof.

205. The method of claim 187, comprising placing a plurality of samples in a microscale device.

206. The method of claim 187, wherein the interaction comprises a binding interaction.

207. The method of claim 187, wherein the detection of the interaction is by at least one or more of a spectrophotometer, fluorometer, photodiode, photomultiplier tube, microscope, scintillation counter, camera, CCD camera, membrane, optical detection system, temperature sensor, conductivity meter, potentiometer, ammeter, pH meter, or combinations thereof.

208. A method of separating materials, the method comprising:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(iv) combinations thereof; and wherein the microfluidic device comprises one or more microscale channels, and wherein at least one of the one or more microscale channels comprises a separation region;

(c) flowing the mixture through the separation zone; and

(d) separating the first material from the second material in the separation region to form at least one separated material.

209. The method of claim 208, wherein the material of at least one layer of the material covers a surface of at least one of the one or more microscale channels.

210. The method of claim 208, wherein the one or more microscale channels comprise an integrated network of microscale channels.

211. The method of claim 209, wherein the microscale channels of the integrated network intersect at a predetermined point.

212. The method of claim 208, wherein the microfluidic device comprises one or more patterned layers of a first polymeric material, and wherein the one or more patterned layers of the first polymeric material define the one or more microscale channels.

213. The method of claim 212, wherein the microfluidic device further comprises a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of the first polymeric material.

214. The method of claim 212, wherein the one or more microscale channels comprise an integrated network of microscale channels.

215. The method of claim 214, wherein the microscale channels of the integrated network intersect at a predetermined point.

216. The method of claim 208, wherein the separation region comprises a functionalized surface.

217. The method of claim 208, wherein the separation region comprises a chromatographic material.

218. The method of claim 217, wherein the chromatographic material is selected from the group consisting of a size separation matrix, an affinity separation matrix, and a gel exclusion matrix or a combination thereof.

219. The method of claim 208, wherein the first or second material comprises one or more constituent members of one or more libraries of chemical or biological compounds or components.

220. The method of claim 208, wherein the first or second material comprises one or more of a nucleic acid template, a sequence reagent, a primer extension product, a restriction enzyme, a PCR reagent, a PCR reaction product, or a combination thereof.

221. The method of claim 208, wherein the first or second material comprises one or more of an antibody, a cellular receptor, an antigen, a receptor ligand, an enzyme, a substrate for an enzyme, an immunochemical, an immunoglobulin, a virus-binding component, a protein, a cytokine, a growth factor, an inhibitor, or a combination thereof.

222. The method of claim 208, comprising detecting the separated material.

223. The method of claim 222, wherein the detection of the separated material is performed by at least one or more of a spectrophotometer, fluorometer, photodiode, photomultiplier tube, microscope, scintillation counter, camera, CCD camera, membrane, optical detection system, temperature sensor, conductivity meter, potentiometer, ammeter, pH meter, or combinations thereof.

224. A method of dispensing material, the method comprising:

(ii) a functionalized PFPE material;

(iii) a fluoroolefin-based elastomer; and

(b) providing at least one material;

(c) placing the at least one material in at least one of the one or more microscale channels; and

(d) dispensing the at least one material through the outlet orifice.

225. The method of claim 224, wherein the at least one layer of material covers a surface of at least one of the one or more microscale channels.

226. The method of claim 225, wherein the one or more microscale channels comprise an integrated network of microscale channels.

227. The method of claim 226, wherein the microscale channels of the integrated network intersect at a predetermined point.

228. The method of claim 224, wherein the microfluidic device comprises one or more patterned layers of the first polymeric material, and wherein the one or more patterned layers of the first polymeric material define the one or more microscale channels.

229. The method of claim 228, wherein the microfluidic device further comprises a patterned layer of a second polymeric material, wherein the patterned layer of the second polymeric material is in operative communication with at least one of the one or more patterned layers of the first polymeric material.

230. The method of claim 228, wherein at least one patterned layer of material comprises a functionalized surface.

231. The method of claim 228, wherein the one or more microscale channels comprise an integrated network of microscale channels.

232. The method of claim 231, wherein the microscale channels of the integrated network intersect at predetermined points.

233. The method of claim 224, wherein the material comprises a drug.

234. The method of claim 233, comprising metering in a predetermined dose of the drug.

235. The method of claim 234, comprising dispensing a predetermined dose of the drug.

236. The method of claim 224, wherein the material comprises an ink composition.

237. The method of claim 236, comprising dispensing the ink composition on a substrate.

238. The method of claim 237, wherein the ink composition is dispensed onto the substrate to form a printed image.