US20100233763A1 - Dual-sided thermal cycler - Google Patents

Abstract

A device, system, and method are provided for thermally treating a fluid processing device. According to various embodiments, a system is provided that can include a thermal device and a fluid processing device holder. The thermal device can include a first block having a thermal conductivity greater than 0.5 Watt per centimeter Kelvin (W/cm·K), a second block having a thermal conductivity greater than 0.5 W/cm·K, and a heat-pump device disposed between the first block and the second block. The heat-pump device can transfer thermal energy from at least one of the first block and the second block to the other of the first block and the second block. The fluid processing device holder can hold a fluid processing device in a heat-transfer position with respect to the first block and the second block. The fluid processing device can be a microfluidic device.

Description

CROSS REFERENCE TO RELATED APPLICATION
• The present application claims benefit under 35 U.S.C. §119(e) from earlier filed U.S. application Ser. No. 12/552,218, filed Sep. 1, 2009, which is a divisional of U.S. patent application Ser. No. 10/926,915, filed Aug. 26, 2004, now U.S. Pat. No. 7,585,663, issued on Sep. 8, 2009, both of which are herein incorporated by reference in their entirety.
FIELD
• The present teachings relate to a thermal energy manipulation system.
BACKGROUND
• There is a need for a device that can provide efficient thermal cycling of a fluid in a retainment region of a fluid processing device while maintaining an adjacent second retainment region at a different temperature. A device compatible with biological or chemical reactions, for example, nucleotide amplification chemistries, and laboratory instrumentation adapted to process a relatively large number of biological/chemical reactions, is desirable.
BRIEF DESCRIPTION
• According to various embodiments, a system is provided that can include a thermal device and a fluid processing device holder. The thermal device can include a first block having a high thermal conductivity, a second block having a high thermal conductivity, and a heat-pump device disposed to transfer thermal energy between the first block and the second block. Herein, high thermal conductivity refers to a thermal conductivity of greater than 0.5 W/cm·K. The heat-pump device can be disposed physically between or adjacent the first block and the second block. The heat-pump device can transfer thermal energy from at least one of the first block and the second block to the other of the first block and the second block. The fluid processing device holder can hold a fluid processing device in a heat-transfer position with respect to the first block and the second block.
• According to various embodiments, a method is provided that can include providing a fluid processing device including at least one fluid processing pathway. Each fluid processing pathway can include at least a first fluid retainment region, a second fluid retainment region, and a fluid communication between the first fluid retainment region and the second fluid retainment region. The method can include providing a thermal device including a first block having a high thermal conductivity, a second block having a high thermal conductivity, and a heat-pump device disposed to transfer thermal energy between the first block and the second block. The method can include transferring thermal energy from at least one of the first block and the second block to the other of the first block and the second block, using the heat-pump device. At the same time, the fluid processing device can be held in a heat-transfer position wherein the first fluid retainment region can be in thermal contact with the first block and the second fluid retainment region can be in thermal contact with the second block. The fluid communication can be sealable, interruptible, closeable, openable, or the like.
• Additional features and advantages of various embodiments will be set forth in part in the description that follows, and in part will be apparent from the description, or can be learned by practice of various embodiments. Other advantages of the various embodiments will be realized and attained by means of the elements and combinations particularly pointed out in the application.
BRIEF DESCRIPTION OF THE DRAWINGS
• Various embodiments of the present teachings are exemplified in the accompanying drawings. The teachings are not limited to the embodiments depicted in the drawings, and include equivalent structures and methods as set forth in the following description and as would be known to those of ordinary skill in the art in view of the present teachings. In the drawings:
• FIG. 1 is a top view of a thermal system according to various embodiments;
• FIG. 2 is a side, partial cross-sectional view of a thermal system including a dual-sided thermal device, according to various embodiments;
• FIG. 3 is an cross-sectional view a thermal system including a single-sided thermal device, a fluid processing device, and a platen, according to various embodiments;
• FIG. 4 is a perspective view of a plurality of blocks positioned with respect to a plurality of linearly-aligned fluid retainment regions of a fluid processing device, according to various embodiments;
• FIG. 5 is a perspective view of a plurality of blocks positioned with respect to a plurality of linearly-aligned fluid retainment regions of a fluid processing device, according to various embodiments;
• FIG. 6 is a bottom view of a fluid processing device positioned over a thermal device, according to various embodiments;
• FIG. 7ais a front view of an exemplary system that can be used to process a fluid processing device;
• FIG. 7bis an exploded view of the system shown inFIG. 7a, in partial phantom, with the top cover removed;
• FIG. 7cis a side view of the device shown inFIG. 7a;
• FIG. 7dis an exploded view in partial phantom of the device shown inFIG. 7cwith the cover open;
• FIG. 7eis an enlarged view of the assembly loading door of the system shown inFIG. 7a;
• FIG. 7fis an enlarged view of a portion of the system shown inFIG. 7a, depicting the positions of the valve actuators, thermal devices, and electronics;
• FIG. 7gis an enlarged view of a section of the system shown inFIG. 7apartially cutaway to show the two-assembly platen;
• FIG. 7his an enlarged view of a section of the system shown inFIG. 7aincluding an assembly loaded in the assembly-loading door;
• FIG. 7iis an enlarged view of a section of the system shown inFIG. 7a, in partial cutaway to show two assemblies loaded for spinning while being held to the rotating platen; and
• FIG. 8 is an enlarged, perspective view of a fluid processing device according to various embodiments;
• FIG. 9 is a perspective view of a fluid processing device according to various embodiments;
• FIG. 10 is a chart of temperatures, plotting the temperatures of a first fluid in a first fluid retainment region and a second fluid in a second fluid retainment region, according to various embodiments; and
• FIG. 11 is a graph that plots the temperatures of five fluid retainment regions, versus time, achieved according to various embodiments.
• It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are intended to provide a further explanation of the various embodiments of the present teachings.
DESCRIPTION OF VARIOUS EMBODIMENTS
• According to various embodiments, a system is provided that can include a thermal device and a fluid processing device holder. The thermal device can include a first block, a second block, and a heat-pump device disposed to transfer thermal energy between the first block and the second block. Herein, high thermal conductivity refers to a thermal conductivity of greater than 0.5 W/cm·K. The heat-pump device can be disposed physically between or adjacent the first block and the second block. The heat pump can be disposed at ends of, or end regions of or otherwise adjacent both the first block and the second block. The heat-pump device can transfer thermal energy from at least one of the first block and the second block to the other of the first block and the second block. The fluid processing device holder can hold a fluid processing device in a heat-transfer position with respect to the first block and the second block.
• Each of the first and second blocks can include a thermally conductive metal material, such as steel or aluminum.
• According to various embodiments, the system can include a control device to control the heat-pump device to provide the first block with a temperature that is greater than the temperature of the second block. The control device can control the heat-pump device to transfer heat from the first block to the second block and to transfer heat from the second block to the first block. The control device can control the temperature of the first block to be at least, for example, about 15° C., about 30° C., or about 40° C., greater than the temperature of the second block. According to various embodiments, the control device can control the heat-pump device to heat the first block to about 95° C., and to keep the temperature of the second block to be less than about 50° C.
• According to various embodiments, at least one of the first block and the second block can include a surface, and the thermal device can further include a thermal interface material disposed in contact with the surface. The first block can include a first heat-transfer surface and the second block can include a second heat-transfer surface, where the first heat-transfer surface and the second heat-transfer surface can be flush with and/or co-planar with one another. According to various embodiments, the heat transfer surfaces of the first block and the second block can be slightly offset from one another. For example, the heat transfer surface of the first block can be positioned relatively raised or offset as compared to the heat transfer surface of the second block. The heat transfer surfaces can lie on planes parallel to one another, but not co-planar. The offset position of the first block heat transfer surface can improve thermal contact between a fluid processing device and the first block heat transfer surface. The offset between the heat transfer surfaces of the first block and the second block can be, for example, from about 0.005 inch to about 0.1 inch, or from about 0.01 inch to 0.020 inches.
• A thermal interface material including a flat surface can be disposed on the heat transfer surface of either or both of the first block and the second block. The thermal interface material can be compressible. In an uncompressed state, thermal interface material provided at the first block can provide a raised surface relative to the heat transfer surface of the first block. In operation, the thermal interface material can be compressed against a portion of a fluid processing device. The thermal interface material can be raised relative to the first block heat transfer surface by from about 0.05 inch to about 0.25 inch, in an uncompressed state. The heat transfer surface of the first block can be offset by about 0.015 inch compared to the heat transfer surface of the second block. When a 0.020 inch thick thermal interface material is disposed against the heat transfer surface of the second block, the raised surface of the thermal interface material can protrude about 0.005 inch or more as compared to the heat transfer surface of the first block. The thermal interface material can comprise a heat conductive material, for example, GAPPAD VO ULTRA SOFT available from Berquist Company, Chanhassen, Minn.
• According to various embodiments, the system can include at least one heat sink in thermal contact with at least one of the first block and the second block. The heat sink can be a metal block with fins or other features providing a large surface area for thermal exchange with another thermal mass, for example, air. The system can include at least one fan adapted to create an air-current in thermal contact with at least one of the first block, the second block, and if a heat sink is included, the heat sink.
• The system can include at least one temperature sensor in thermal contact with at least one of the first block and the second block. The system can include a pressing device adapted to force at least one of the first block and the second block into thermal contact with a fluid processing device when such a fluid processing device is held by the fluid processing device holder. The system can include an alignment device to operatively position the fluid processing device holder and the heat-pump device with respect to one another.
• According to various embodiments, the system can include a platform and a second thermal device supported by the platform. The second thermal device can include a third block, a fourth block, and a second heat-pump device disposed between the third block and the fourth block. The second heat-pump device can transfer thermal energy from at least one of the third block and the fourth block to the other of the third block and the fourth block. The first block, the second block, the third block, and the fourth block can be disposed in respective heat-transfer positions. According to various embodiments, the first block, the second block, the third block, and the fourth block can be simultaneously disposed in respective heat-transfer positions.
• According to various embodiments, the thermal device can include a third block and a second heat-pump device disposed between the first block and the third block, where the second heat-pump device can transfer thermal energy from at least one of the first block and the third block to the other of the first block and the third block. The system can include a control device that can control the heat-pump device to provide the first block with a temperature that can be greater than the temperature of the third block. The fluid processing device holder can hold a fluid processing, for example, a microfluidic card-type device.
• According to various embodiments, the system can include a fluid processing device held by the fluid processing device holder. The fluid processing device can include at least one fluid processing pathway. Each fluid processing pathway can include at least a first fluid retainment region, a second fluid retainment region, and a fluid communication between the first fluid retainment region and the second fluid retainment region. The first fluid retainment region can be positioned in a heat-transfer position with respect to the first block of the system, and the second fluid retainment region can be positioned in a heat-transfer position with respect to the second block of the system. The system can include a fluid movement device, unit, or system, adapted to move or transfer a fluid from the first fluid retainment region to the second fluid retainment region via the fluid communication. The system can include a valve manipulation device supported by a common or a different platform, and the fluid processing device can include a valve between the first fluid retainment region and the second fluid retainment region. The at least one fluid processing pathway can include a plurality of fluid processing pathways each including a respective first fluid retainment region and a respective second fluid retainment region. The plurality of first fluid retainment regions can be aligned linearly with respect to each other. The plurality of first fluid retainment regions can be evenly or differently spaced from adjacent first fluid retainment regions. According to various embodiments, the second fluid retainment region can include a plurality of second fluid retainment regions aligned linearly with respect to each other, and evenly or differently spaced from one another.
• According to various embodiments, a method is provided that can include manipulating a fluid processing device that includes at least one fluid processing pathway, wherein the pathway includes at least a first fluid retainment region, a second fluid retainment region, and a fluid communication between the first fluid retainment region and the second fluid retainment region. The method can include providing a thermal device. The thermal device can include a first block, a second block, and a heat-pump device in thermal contact with, for example, disposed between, the first block and the second block. The method can include transferring thermal energy from at least one of the first block and the second block to the other of the first block and the second block using the heat-pump device, and holding the fluid processing device in a heat-transfer position. The first fluid retainment region can be in thermal contact with the first block and the second fluid retainment region can be in thermal contact with the second block.
• According to various embodiments, the method can include controlling the heat-pump device to provide the first block with a temperature that can be greater than the temperature of the second block. The controlling can include heating the first block to an elevated temperature, for example, to a temperature of about 95° C. The method can include maintaining the temperature of the second block to be less than the elevated temperature, for example, less than about 50° C. when the elevated temperature exceeds 50° C. The controlling can include achieving and maintaining the temperature of the first block to be at least about 40° C. greater than the temperature of the second block. The controlling can include inverting a direction of the thermal transfer with respect to the first block and the second block. The controlling can include controlling the heat-pump device to effect heat transfer from the first block to the second block and/or from the second block to the first block.
• According to various embodiments, the method can include heating the first block with a heat-generating device. According to various embodiments, the method can include creating an air-current in thermal contact with at least one of the first block and the second block. According to various embodiments, the method can include cycling a temperature of the first block from a first temperature to a second temperature, and then back to the first temperature. According to various embodiments, the method can include forcing at least one of the first block and the second block into thermal contact with a fluid processing device. According to various embodiments, the method can include positioning the fluid processing device and the thermal device with respect to one another.
• According to various embodiments, the method can include providing a second thermal device including a third block, a fourth block, and a second heat-pump device disposed to transfer heat between the third block and the fourth block. The second heat pump can be disposed physically between, at ends of or end regions of or otherwise adjacent, the first block and the second block. The method can include transferring thermal energy from at least one of the third block and the fourth block to the other of the third block and the fourth block, and positioning the second thermal device such that or wherein the third block can be in a heat-transfer position with respect to the first fluid retainment region, and the fourth block can be in a heat-transfer position with respect to the second fluid retainment region.
• According to various embodiments, the method can include providing the thermal device with a third block and a second heat-pump device disposed to transfer heat between the first block and the third block. The second heat-pump device can be adapted to transfer thermal energy from at least one of the first block and the third block to the other of the first block and the third block. The first block can be arranged or disposed, for example, between the second block and the third block.
• According to various embodiments, the method can include loading a first set of materials reactive at a first minimum temperature into the first fluid retainment region and loading a second material into the second fluid retainment region. The second material can include a material that can be rendered substantially ineffective when subjected to the first minimum temperature. The first set of materials can include nucleic acid amplifying reagents. The nucleic acid amplifying reagents can include polymerase chain reaction reagents. The first set of materials can include oligonucleotide ligase reaction reagents. The second material can include size-exclusion ion-exchange media. The second material can include reagents capable of performing purification, for example, by filtering and/or by ion-exchange.
• According to various embodiments, the method can include loading a target nucleic acid material and amplifying reagents into the first fluid retainment region and amplifying the nucleic acid material by thermal cycling in the first fluid retainment region. The method can include loading size-exclusion ion exchange media into the second fluid retainment region. The method can include loading reagents for a purification reaction into the second fluid retainment region, for example, ingredients for an ion-exchange reaction.
• According to various embodiments, the temperature of a fluid retainment region in a fluid processing device can be controlled by a thermal cycler that includes a Peltier-effect thermoelectric device, a heat-pump, an electrical resistance heating element (Joule heater), fluid flow through channels in a metal block, reservoirs of fluid at different temperatures, a tempered air impingement device, or a combination thereof. According to various embodiments, a plurality of blocks can be maintained at various temperatures to maintain different temperatures in different fluid retainment regions of a fluid processing device. The different temperatures can be maintained in a plurality of areas separated by imaginary lines drawn between a plurality of fluid retainment regions in a fluid processing device. According to various embodiments, a plurality of blocks can be aligned with the temperatures as desired. Thermal energy can be transferred between the blocks. Thermal energy can be transferred from the blocks to the fluid processing device.
• According to various embodiments, cycling of a block temperature between various temperatures can effect quick changes in the temperature of a fluid retainment region. More than one heat pump can be disposed between the blocks to increase the heat pumping capacity of the system, allowing for faster cooling and/or heating of the blocks. After achieving a desired temperature for a block, a temperature sensor can be used to maintain the desired temperature of a fluid retainment region to be within about +/−5° C. of a desired temperature, for example, to be within about +/−1° C., or about +/−0.5° C. A computer system can be used to control the thermal system. A computer system can be used to maintain a desired temperature for a desired time period and for maintaining a desired number of cycles. Blocks can be placed adjacent one another, without contacting one another, to achieve and maintain the blocks at disparate temperatures. A thermal insulator can be placed between two adjacent blocks to help maintain the two blocks at different respective temperatures. A thermal insulator can be placed between two adjacent blocks to maintain the two blocks at the same temperature.
• According to various embodiments, the system can include a thermal device in thermal contact with a fluid retainment region of a fluid processing device. The fluid retainment region can be included in a substrate, for example, a substrate including a first and second surface. The thermal device can be disposed along the first or the second surface, for example, above or below the fluid retainment region of the fluid processing device. Such a configuration of a thermal device is herein referred to as a single-sided thermal cycler.
• According to various embodiments, the system can include two thermal devices aligned to regulate the temperature of a fluid retainment region, where one of the two thermal devices is disposed along a first surface of a fluid processing device and the other of the two thermal devices is disposed along an opposite, second surface of the fluid processing device. Such a configuration of thermal devices is referred to herein as a double-sided thermal cycler.
• According to various embodiments, any of the blocks described above can be shaped as a disk, a cylinder, a cuboid, a rectanguloid, a bar, or as any other shape. In the accompanying drawings, and for the sake of simplicity, a rectanguloid block is depicted. A surface of the block can be designed to complement a region of a fluid processing device that is to be thermally regulated. Shapes of the blocks used herein can be altered to match a shape of a thermally regulated region of a fluid processing device. The shape of the block can include a flat surface to thermally contact a heat-transfer surface of a heat-pump, for example, a heat absorbing surface of a thermoelectric device.
• With reference to the drawings,FIG. 1 is a top view of athermal system160. Thethermal system160 can include acarriage152 that is capable of sliding along rails in a plurality of directions. Thecarriage152 can be capable of latitudinal and/or longitudinal movements.Rails154 and156 can be included to provide guidance for the latitudinal and/or longitudinal movements. According to various embodiments, thecarriage152 can be permanently affixed, for example, to a base or platform, such as tobase150 as shown. Afirst block102, can be configured as a relatively hot block, and can be thermally coupled to one or morethermoelectric device130,131. Thethermoelectric device130,131 can be thermally coupled to asecond block104 and to athird block105, that can be configured to be relatively cold blocks. One or more alignment pins103 can be disposed at or near an end of thefirst block102. The alignment pins103 can align thefirst block102 with a fluid retainment region such as a reaction well (not shown) formed in or on a fluid processing device (not shown). Heat sinks106,107, for example, each including a plurality of cooling fins, can be in thermal contact with thesecond block104 and thethird block105. Afan155 can be provided to create an air flow in the direction indicated to thus flow over and cool theheat sink106. Afan153 can be provided to create an air flow in the direction indicated to thus flow across and cool theheat sink107. Thethermoelectric device130,131 can be thermally coupled to and disposed between thefirst block102 and thesecond block104, as shown at130, and between thefirst block102 and thethird block105, as shown at131. According to various embodiments, a plurality of thermoelectric devices like130 and131 can be thermally coupled to at least thefirst block102.
• FIG. 2 is a side, partial cross-sectional view of athermal system100 including twothermal devices140 and142, that together form a dual-sided thermal cycler. The twothermal devices140,142 can be made to be in thermal contact with afluid processing device120. Eachthermal device140,142 respectively includes afirst block102, athermoelectric device130, asecond block104, athird block105, and twoheat sinks106. Eachthermal device140,142 can be assembled using articles known in the art, for example,bolts132. Eachsecond block104 can include athermal interface material124. Thethermal interface material124 can be a compressible material. A second thermal interface material (not shown) can be disposed on thefirst block102. Thefluid processing device120 can include a firstfluid retainment region121 and a secondfluid retainment region122. The firstfluid retainment region121 can be subjected to higher temperatures than the secondfluid retainment region122. Thefirst block102 can include a heat-transfer surface114 that can be aligned into a heat-transfer position with respect to the firstfluid retainment region121 of thefluid processing device120. Thethird block105 can include a heat-transfer surface110 that can be aligned into a heat-transfer position with respect to the secondfluid retainment region122. Theheat transfer surface114 can be co-planar or flush with the heat-transfer surface110. Theheat transfer surface114 can be offset from the plane of the heat-transfer surface110. Thethermal interface material124 can provide a heat-transfer surface that is co-planar with the heat-transfer surface124. Thefluid processing device120 can be compressed between thethermal devices140 and142 using apressing assembly144. Thepressing assembly144 can include, for example, a clamping device, a set of bolts, a pneumatic press, a hydraulic press, or a multiplicity or combination thereof. Thepressing assembly144 can include a device that is capable of moving either thefluid processing device120 or the heat-transfer assembly, in relation to one another, and between a pressure-application position and a release position.
• According to various embodiments, the thermal system can include two thermal devices, including a lower thermal device and an upper thermal device. The lower thermal device can include a hot block sandwiched between two thermoelectric devices and two cold blocks, each thermal device sandwiching a respective thermoelectric device between the respective cold block and the hot block. The upper thermal device can be identical to and/or a mirror image of, the lower thermal device. The upper thermal device can be disposed above the lower thermal device, as a mirror or as an inverted version of, the lower thermal device. According to various embodiments, during the heating portion of a thermal cycle, the thermoelectric devices, in both the upper and lower thermal devices, can transfer heat from the upper and lower flanking cold blocks to the respective upper and lower hot blocks. According to various embodiments, during the cooling portion of the thermal cycle, the direction of the heat transfer by the upper and lower thermoelectric devices can be reversed, and the upper and lower thermoelectric devices can transfer heat from the upper and lower hot blocks to the respective upper and lower flanking cold blocks.
• According to various embodiments, a fluid processing device including at least one plurality of linearly aligned fluid retainment regions can be positioned between the upper and lower thermal devices whereby the plurality of linearly aligned fluid retainment regions can be disposed centered on the hot block of at least one of the upper and lower thermal devices. A fluid retainment region in the at least one plurality of linearly aligned reaction reactions can be used for, for example, a nucleotide amplification reaction, a nucleotide sequencing reaction, an oligonucleotide synthesis reaction, or the like. According to various embodiments, the hot blocks of the upper and lower assemblies can clamp the fluid processing device therebetween. According to various embodiments, the cold blocks of the upper and lower assemblies can clamp the fluid processing device therebetween. According to various embodiments, the pressing device can include the lower thermal device and the upper thermal device to press against the fluid processing device during operation. The pressing device and the two thermal devices can operate at the same time or at different start and/or stop times.
• According to various embodiments, the heat capacity (Joules/Kelvin) of the cold blocks can be larger than the heat capacity of the hot blocks, for example, about 5-times, about 10-times, or about 50-times. The cold blocks can use the surrounding environment, for example, air, as a heat source or as a heat sink depending on the direction of heat-transfer by the thermoelectric device, by using heat sinks thermally coupled to the cold blocks. The control device can control the temperature of the hot block to be at least about 10° C., at least about 15° C., at least about 20° C., at least about 30° C., at least about 40° C., or at least about 50° C., greater than the temperature of the cold block.
• According to various embodiments, the terms “cold block” and “hot block” exemplify the relative temperatures of the blocks relative to one another. The cold block can be hotter than the ambient environment. The hot block can be cooler than the ambient environment. In operation, the hot block can be hotter than the cold block. This can occur, for example, when the hot block is heated to a temperature of about 95° C. and the cold block is heated to a temperature of only about 50° C. or less. In operation, the hot block can be cooler than the cold block. This can occur, for example, when after completion of a heating cycle the hot block is cooled to a temperature of about 25° C. while the cold block maintains or reduces to a temperature of above about 25° C.
• According to various embodiments, each thermal device can include a fan to direct air-flow over heat sinks and/or a cold block to assist in a heat transfer between blocks. According to various embodiments, the thermal conductivity of a fluid processing device can be less than the thermal conductivity of a block. According to various embodiments, the thermal conductivity of the block can be greater than the thermal conductivity of an ambient environment, for example, air at about one atmosphere of pressure at about 300° K.
• The thermal conductivity of a material is equivalent to the quantity of heat that passes in a unit time through a unit area of a plate, when the material's opposite faces are subject to a unit temperature gradient, such as, a one degree temperature difference across a thickness of one unit. Thermal conduction is the transfer of heat or thermal energy from one substance to another.
• According to various embodiments, the block or bar can include a material having a high thermal conductivity, for example, diamond or metal, for example, aluminum, copper, steel, stainless steel, alloys or combinations thereof. At temperatures of about 273° K. to about 300° K. various materials having a typical thermal conductivity of at least about 0.5 Watts per centimeter Kelvin (W/cm·K), for example, at least about 1.9 W/cm·K, or at least about 5 W/cm·K, can be used to form a block. According to various embodiments, a block can be a high thermal conductivity block when the block has a thermal conductivity of at least about 0.5 W/cm·K, for example, at least about 1.9 W/cm·K, or at least about 5 W/cm·K. at temperatures of about 273° K. to about 300° K.
• According to various embodiments, a heat-pump can be a machine, that moves heat from a low level of temperature to a higher level of temperature under supply of work, for example, gas compression heat-pumps, phase change heat-pumps, or thermoelectric heat-pumps that use the Peltier effect. A heat-pump can include a vapor-cycle device, for example, a Freon-based vapor compression or absorption refrigerator. Vapor-cycle devices can include moving mechanical parts and require a working fluid, while thermoelectric elements can be totally solid state.
• According to various embodiments, a thermoelectric device can be used as a heat-pump, for example, an XLT module available from Marlow Industries, Inc. of Dallas, Tex. Controls for a thermoelectric device can include an adjustable-bipolar DC output current power supply. The power supply can provide programmable PID control/ramp set points to control the thermoelectric device, deviation alarms, and automatic and manual operating modes. In reactions, for example, a Polymerase Chain Reaction (PCR), thermoelectric devices can both heat and cool fluid retainment regions at a predetermined rate by using a bi-directional power supply under computer control. Thermoelectric devices can be specifically designed to withstand the continuous temperature excursions required in PCR use.
• According to various embodiments, a thermoelectric device can include a heat absorbing surface. The thermoelectric device can include a heat dissipating surface. The heat absorbing surface (cooling surface) can be in thermal contact with a cool block. The heat dissipating surface (heating surface) can be in thermal contact with a hot block. A control device can reverse the polarity of a voltage being applied to the thermoelectric device to reverse a direction of the heat flow effectively swapping the heat absorbing surface with the heat dissipation surface. When power is supplied to a thermoelectric device, the heat absorbing surface and the heat dissipating surface are activated, and each surface can be referred to as an active surface.
• According to various embodiments, a thermal interface material (TIM) can provide a good thermal contact between two surfaces. The TIM can include silicone-based greases, elastomeric pads, thermally conductive tapes, thermally conductive adhesives, or a combination thereof. Zinc-oxide silicone can be a common TIM. An elastomeric pad compressible to about 90% of an uncompressed thickness under an exerted pressure of at least 0.5 pounds per square inch (PSI), for example, at least about one (1) PSI, at least about five (5) PSI. According to various embodiments, Gap-Pad products, for example, the GAP PAD VO ULTRA SOFT available from Berquist Company of Chanhassen, Minn. can be used as a thermal interface material. A TIM is described in U.S. Pat. No. 5,679,457 to Bergerson, which is incorporated herein in its entirety.
• According to various embodiments, a thermal connect, a thermal contact, or a thermal conduct can be a physical contact of two objects, for example, a fluid processing device surface with a block surface. According to various embodiments, a TIM can be disposed between the fluid processing device surface and the block surface, on the fluid processing device surface, or on the block surface to optimize the thermal contact. According to various embodiments, the block surface can be pressed against the fluid processing device surface to optimize the thermal contact.
• According to various embodiments, a thermal insulator can provide low thermal conductivity. A thermal insulator can be used to prevent heat transfer from a temperature regulated volume to a temperature unregulated volume, for example, a thermal insulator can be used to prevent heat transfer from a heated volume to a cooled or unregulated volume. A thermal insulator can include wood, plastics, air, or gases. According to various embodiments, a thermal insulator can be disposed between or adjacent a heated fluid retainment region in a fluid processing device, and the fluid processing device holder or platen holding the fluid processing device.
• According to various embodiments, a pressing device surface can exert a pressure against a first or operative surface of a fluid processing device. The pressing device surface can be provided by a thermal device. According to various embodiments, the pressing device surface can press the fluid processing device across different imaginary zones or regions of the fluid processing device. The pressing device surface can apply a compression force of, for example, from about one (1) lbs to 500 lbs, about 10 lbs, about 20 lbs, about 50 lbs, about 100 lbs, or about 200 lbs. The average force over the thermal cycled area can be approximately 20 lbs per square inch. Pressure exerted over a fluid retainment region can prevent leakage of a fluid retained in the fluid retainment regions. A single-sided thermal device can press against the fluid processing device. A dual-sided thermal device can press against the fluid processing device. A fluid processing device holder or platen can press against the fluid processing device.
• According to various embodiments, a fluid processing device can move relative to a thermal device by shifting the fluid processing device, shifting the thermal device, or shifting both the fluid processing device and the thermal device. The shifting can be performed by placing either the fluid processing device or the thermal device on a movable fluid processing device holder. The fluid processing device holder can be included or disposed in a carriage. The fluid processing device holder can be movable by a robotic arm, other devices well known in the art, or the fluid processing device can be manually transferred. The fluid processing device holder can move along a single-axis, two-axes, or all three-axes. According to various embodiments, the fluid processing device and/or the thermal device can move to heat or cool a plurality of fluid retainment regions formed in or on a fluid processing device as desired. According to various embodiments, a platen can be spun to align a desired fluid retainment region with a heat-transfer position with respect to a thermal device. According to various embodiments, the fluid processing device holder can be used to force the fluid processing device and/or the thermal device against one another to provide a pressing force. According to various embodiments, the fluid processing device holder can be moved to allow another system, for example, a valve manipulation system, a fluid depositing system, an output retrieval system, a fluid movement system, a detection system, or a combination thereof, to access the fluid processing device.
• According to various embodiments, an alignment system can generate a signal upon detecting a fluid processing device in a heat-transfer position. A control system can begin a thermal cycle based on the signal. The signal can be generated by using, for example, optical alignment, mechanical alignment, electrical alignment, manual alignment, or an on/off switch. According to various embodiments, alignment pins included in the thermal device can be solenoids. According to various embodiments, the alignment system can indicate an error in the fluid processing device placement by generating an error signal, for example, by beeping, by lighting an error light, or by indicating an alignment error on a computer screen.
• According to various embodiments, a fluid processing device can include an alignment device, for example, a hole, a notch, a pin, a chamfered edge, a chamfered corner, or combinations thereof. The alignment device can assist in the correct placement of the fluid processing device in the fluid processing device holder. According to various embodiments, a fluid processing device holder can include an alignment device, for example, a hole, a notch, a pin, a chamfered edge, a chamfered corner, for correct operative alignment of the fluid processing device holder with respect to a thermal device. A plurality of exemplary alignment devices can be disposed in or on the fluid processing device and/or the fluid processing device holder. For example, two alignment pins in the fluid processing device holder can align a fluid processing device into a heat transfer position wherein the fluid device can include two holes or notches.
• According to various embodiments, a thermal device can supplement heat from one or more thermoelectric devices by using a heat generating device, for example, a heat resistive wire. The heat generating device can be eliminated to reduce cost. According to various embodiments, a temperature sensor can include, for example, a thermometer, a thermistor, or a thermocouple sensor.
• According to various embodiments, a region of the fluid processing device holder operatively in a good or high thermal contact with a fluid retainment region can include a heating element, for example, a thin-resistive heater. During a thermal cycling protocol, the heating element can raise and maintain the temperature of the reaction region at a fixed or uniform temperature, for example, about 40° C., about 50° C., or about 55° C., while the thermal device can be used to oscillate the temperature of the reaction region from about 60° C. to about 95° C. The heating element can reduce the thermal gradient of the fluid processing device. This reduction in the thermal gradient due to the use of a heating element can be greater than a reduction in the thermal gradient when a thermal insulator is operatively disposed in good or high thermal contact with the fluid retainment region. The heating element can be used in conjunction with a thermal insulator. The heating element can have a low mass. The heating element can have a long lifetime. The heating element can be of durable construction.
• According to various embodiments, a thermal device can provide one or more thermoelectric devices that can transfer heat between two blocks. According to various embodiments, one, two, or more thermal devices can surround a fluid processing device. According to various embodiments, a thermal system can provide one, two, or more thermal assemblies, wherein a thermal assembly is one or more thermal devices configured to operate cooperatively on a fluid processing device disposed in a fluid processing device holder. The thermal assemblies in a thermal system can correspond to the number of fluid processing device holders in a thermal system. The dual-sided thermal system described above is an example of a thermal assembly including two thermal devices.
• According to various embodiments, a thermal cycling system using a single-sided thermal device can reduce the mechanical complexity of the thermal cycling and platen sub-assemblies. The single-sided thermal device can lead to a fluid processing device holder or platen design that can be more aerodynamic and structurally sound.
• FIG. 3 a side partial cross-section of athermal system601 including athermal device600 and aplaten630. Thethermal device600 can include a plurality ofthermoelectric devices612,622,624,628. The plurality ofthermoelectric devices612,622,624,628 can be thermally coupled to afluid processing device620. Thethermal device600 can include a first block610. The first block610 can be used to thermally regulate a fluid retainment region635 of thefluid processing device620. The first block610 can be a cold block. A cooling fan609 can direct an air flow across the first block610. The thermoelectric device612 can include a heat absorbing surface613 in contact with the first block610, and can be disposed adjacent or between the first block610 and asecond block606. Thesecond block606 can be used to thermally regulate afluid retainment region633 of thefluid processing device620. Thesecond block606 can be a hot block. Thethermoelectric device622 can include a heat absorbing surface623 in contact with a third block602, and can be disposed adjacent or between thesecond block606 and the third block602. The third block602 can be used to thermally regulate afluid retainment region634 of thefluid processing device620. The third block602 can be a cold block. A cooling fan (not shown) can be disposed to direct an air flow across the third block602. Thethermoelectric device624 can include a heat absorbing surface625 in contact with the third block602, and can be disposed adjacent or between the third block602 and a fourth block604. The fourth block604 can be used to thermally regulate afluid retainment region632 of thefluid processing device620. The fourth block604 can be a hot block. Thethermoelectric device628 can include aheat absorbing surface629 in contact with afifth block608, and can be disposed adjacent or between the fourth block604 and thefifth block608. The fifth block602 can be used to thermally regulate afluid retainment region631 of thefluid processing device620. The third block602 can be a cold block.
• Thefluid processing device620 can be a microfluidic device, for example, themicrofluidic device500 as illustrated inFIG. 8, or the microfluidic device800 ofFIG. 9. Thefluid processing device620 can be placed in a fluidprocessing device holder638 that is disposed in theplaten630. The fluidprocessing device holder638 can be a recess in theplaten630. Thefluid processing device620 can be secured in, on, or to the fluidprocessing device holder638 using locks tabs, adhesive, tape, friction, or other devices well known in the art.Fluid retainment regions632,633 can be subjected to higher temperatures thanfluid retainment regions631,634,635. The fluid processing device holder can include achannel636,637. Thechannel636,637 can complement and align with a shape of a desired fluid retainment region. Thechannel636,637 can have a thermal insulator disposed therein. Theplaten630 can be rotatable.
• Thechannel636,637 can include a heating element (not shown) disposed therein. In operation, acontrol unit640 can use the heating element included in thechannel636,637 to add heat to thefluid processing device620. The added heat can regulate the temperature of fluids contained influid retainment regions632,633.
• Thecontrol unit640 can transfer heat fromblocks602,608,610 toblocks604,606 usingthermoelectric devices612,622,624,628. Thecontrol unit640 can reverse a polarity of a voltage supplied to thethermoelectric devices612,622,624,628 to reverse a direction of the thermal energy transfer, i.e., to transfer heat fromblocks604,606 toblocks602,608,610. Thecontrol unit640 can use a signal from atemperature probe644. Thetemperature probe644 can be a plurality of temperature probes. Thetemperature probe644 can include one or more temperature probes measuring temperatures across a block, for example, blocks602,604,606,608,610. Thecontrol unit640 can independently operatethermoelectric devices612,622,624,628. Thecontrol unit640 can control a movement of thethermal device600 to place thethermal device600 in thermal contact with thefluid processing device620, for example, between a pressure application position and a pressure release position. Arrows inFIG. 3 illustrate possible directions of movement for thethermal device600. Thecontrol unit640 can control apressing device642 to press thethermal device600 against thefluid processing device620.
• According to various embodiments, a system can include a plurality of thermal devices to regulate temperatures of a plurality of fluid retainment regions. The plurality of fluid retainment regions can be disposed in one substrate. The plurality of fluid retainment regions can be disposed in a plurality of substrates.
• According to various embodiments, the thermal device, single-sided or dual-sided, can be very effective in performing many types of chemical reactions requiring various thermal protocols. According to various embodiments, the thermal device can be used for a plurality of reactions: amplification reactions, for example, PCR or Oligonucleotide Ligation Assay (OLA); labeling reactions, for example, sequencing and SnaPshot; or enzymatic purifications, for example, by using Exol, EXO-SAP-IT, or Lambda-Exo nuclease (Lambda-Exo). Exol or Exolnuclease I, and Lambda-Exonuclease are available from New England Biolabs of Beverly, Mass. (see www.neb.com). EXO-SAP-IT is a registered trademark of and available from USB Corporation of Cleveland, Ohio (see www.usbweb.com). SnaPshot is a trademark of and available from Applied Biosystems of Foster City, Calif. (see www.appliedsystems.com). The chemical reactions can be completed in microfluidic devices. The chemical reactions can utilize a dual sided vertical Peltier thermal cycler.
• According to various embodiments, a fluid retainment region can be pre-loaded with a reagent. The pre-loaded reagents can be, for example, dried-down probes, primers, tags, labels, reactants, buffers, or other reagents useful in nucleic acid sequence amplification, sequencing, ligation, or the like, reactions as known to those of ordinary skill in the art. Examples of reagents that can be used and/or preloaded include: for example, reducing agents (e.g. dithiothreitol, tris[2-carboxyethyl]phosphine hydrochloride), alkylating agents (e.g. iodoacetic acid, iodoacetamide), derivatization reagents (e.g. N-hydroxysuccinamide activated biotin, sulfosuccinimidyl-4-O-[4,4′-dimethoxytrityl]butyrate, 2-iminothiolane hydrochloride, fluorescein isothiocyanate), cross-linking reagents (e.g. dimethyl pimelimidate hydrochloride, disuccinimidyl suberate, 1-ethyl-3-[3-dimethylaminopropyl]-carbodiimide hydrochloride), lyophilized enzymes (e.g. trypsin, chymotrypsin, pepsin, papain), immobilized enzymes (e.g. immobilized trypsin), catalysts, enzyme inhibitors, enzyme substrates, dyes, immobilized reagents, or combinations thereof.
• According to various embodiments, the system can include one or more fluid processing device holders, each fluid processing device holder adapted to support a fluid processing device including a reaction region. A system can include a plurality of fluid processing device holders, for example, one, two, four, six, or six or more. The system can include a plurality of thermal devices, for example, one, two, four, six, or six or more. The system can include thermal devices equal in number to the number of fluid processing device holders included in the system. The system can include thermal devices less in number than the number of fluid processing device holders included in the system. The plurality of fluid processing devices disposed in the one or more fluid processing device holders can be thermally regulated by the system in a desired order.
• According to various embodiments, the system can include a platen to support a fluid processing device. The platen can be divided into platen regions, for example, two, four, six, eight, or eight or more. Each platen region can include a fluid processing device holder. The platen regions can be geometric or non-geometric divisions of a plane. The platen can include a first surface. Fluid processing device holders and fluid processing devices therein can be distributed evenly or irregularly in the plurality of platen regions.
• According to various embodiments, the platen can limit thermal variation through the thickness of the microfluidic device. A portion of the platen region can thermally contact the reaction region of the fluid processing device. The reaction region can be in thermal contact with a hot region of the thermal device. The portion of the platen region in thermal contact with the reaction region can include a thermal insulator, for example, black-glass-filled polycarbonate, plastics, air, a polyamide blanket, or gas. The thermal insulator can be moveably disposed in thermal contact with the reaction region. The thermal insulator, a material or material composition with poor thermal conductivity, can be included in the platen region, for example, in a groove in the platen, a circular groove in the platen, a groove complementing the form of a reaction region, or a groove complementing an surface area of a plurality of reaction regions on a fluid processing device. The thermal insulator could be included in the platen such that pockets of air are disposed in thermal contact with the reaction region.
• FIG. 4 is a perspective schematic10 of a plurality of blocks positioned with respect to a plurality of linearly-aligned fluid retainment regions. A first block24 can be in thermal contact with a plurality of fluid retainment regions42, for example, aligned in a linear fashion. A first gap28 can be disposed between the first block24 and a second block22. The second block22 can be disposed to conduct thermal energy to/from a plurality offluid retainment regions31,33,35,37. A second gap29 can be disposed between the second block22 and a third block26. The third block26 can be disposed to conduct thermal energy to/from a plurality offluid retainment regions45, for example, aligned in a linear fashion. The fluid retainment regions can be formed in or on a fluid processing device20. Non-thermally regulated fluid retainment regions41 can also be disposed in the fluid processing device20. Thermoelectric devices (not shown) can be disposed in the first gap28 and/or the second gap29. A control device (not shown) can control the thermoelectric devices to achieve and maintain various temperatures for the first, second and third blocks24,22,26. The blocks can in turn achieve and maintain various temperature zones, zones formed by imaginary lines sectioning the fluid processing device20, and the wells contained in the temperature zones. According to various embodiments, the control device can transfer heat among the first block24, the second block22, and the third block26, with the second block22 acting as an intermediate temperature zone.
• FIG. 5 is a perspective schematic50 of a plurality of blocks positioned with respect to a plurality of linearly-aligned fluid retainment regions. A first block64 can be in thermal contact with a plurality of fluid retainment regions82. A second block62 can be disposed to conduct thermal energy to/from a plurality offluid retainment regions71,75,77. A third block66 can be disposed to conduct thermal energy to/from a plurality offluid retainment regions85. The fluid retainment regions can be formed in or on a fluid processing device60. The blocks can bring various regions or sections of the fluid processing device50 to different temperatures, and maintain the regions at those temperatures. Non-thermally regulated blocks92,94,96 can assist in cooling and/or heating of respective fluid retainment region in thermal contact with the first block64, the second block62, and the third block66. Each non-thermally regulated block92,94,96 can include a high conductivity material, for example, if the fluid retainment region in thermal contact with the non-thermally regulated block92,94,96 is to be kept cooler than an adjacent fluid retainment region. Each non-thermally regulated block92,94,96 can complement the functionality of the opposing first block64, second block62, and third block66, respectively. For example, if the first block64 and the third block66 are to be used as cold blocks, the non-thermally regulated blocks94,96 can include a low conductivity material or a thermal insulator when, for example, the fluid retainment region in thermal contact with the non-thermally regulated block94,96 is to be kept warmer or hotter than an adjacent fluid retainment region. One or more cooling fans (not shown) can be disposed to create an air flow in thermal contact with the non-thermally regulated blocks94,96. The non-thermally regulated block92 can be disposed opposite a hot block (second block62), and can include a thermal insulator such as plastic.
• Each non-thermally regulated block92,94,96 can reduce a thermal load on a heat-pump or a heating element. The non-thermally regulated block92,94,96 can be integrally formed by using a single-piece of thermal insulation to control the thermal load. The single-piece of thermal insulation can include a thermal insulation pad, for example, a polyamide blanket. According to various embodiments, the control device can use the thermoelectric devices to transfer heat among the first block64, the second block62, and the third block66, with the second block62 acting as an intermediate temperature zone.
• FIG. 6 depicts afluid processing device566 thermally coupled with athermal device550 including a first cold block552 and a second cold block556 arranged around ahot block554. A firstthermoelectric device557 can be disposed between the cold block552 and thehot block554. A secondthermoelectric device559 can be disposed between the cold block556 and thehot block554. Thefluid processing device566 can be aligned with thethermal device550 by disposing analignment pin560 in analignment notch562. Thealignment notch562 can be included in thefluid processing device566. Afluid retainment region570 can be thermally coupled to thehot block554, whilefluid retainment regions568,572 can be thermally coupled to the first cold block552 and the second cold block556, respectively. Fluid retainment region574 is not thermally regulated. Afluid input manifold551 can distribute a fluid and/or reagents to thefluid retainment region568. Asecond alignment notch564 can be disposed in thefluid processing device566. According to various embodiments, thefluid processing device566 can be thermally coupled with thethermal device550. By disposing thepin560 in thesecond alignment notch564, thefluid retainment region572 can be thermally coupled with the cold block552 and the fluid retainment region574 can be thermally coupled with thehot block554. Analignment pin558 can be disposed in a separate alignment notch (not shown) in thefluid processing device566.
• FIGS. 7athrough7idepict a system according to various embodiments. Thesystem410 includes anelectronics unit412, arotating platen414, athermal assembly416, acover418, and anenclosure basin420. Thedevice410 also includes an assembly processing unit370 shown inFIGS. 7e-7h.
• The assembly processing component370 includes atray loading door372,electronics412, avalve actuator376, and twothermal assemblies377 and378. Eachthermal assembly377,378 can include two thermal devices. Each thermal device can include a hot block and at least one cold block. The component370 shown particularly inFIG. 7gincludes a two-assembly platen380 for processing two assemblies simultaneously. The non-labelled arrows shown inFIG. 7idepict the direction of centripetal force applied to the assembly resulting from rotation of theplaten380 about acentral axis386 thereof.FIG. 7hshowstray loading door372 in an open position and anassembly381 loaded in the door and ready to be supported by the two-assembly platen380 upon closure of theloading door372.
• FIG. 8 is an enlarged, perspective view of amicrofluidic device500 according to various embodiments that can be used to manipulate fluids, for example, micro-sized fluid samples. Themicrofluidic device500 can include asubstrate510 that can include a plurality of fluid-containment features or fluid retainment regions formed therein or thereon, for example, a plurality offluid retainment regions514,516,518,520,522,524. Thefluid retainment regions514,516,518,520,522,524 can be formed in or on themicrofluidic device500. Other fluid-containment features, for example, reservoirs, recesses, channels, vias, appendices, input wells and ports, output wells, purification columns, or valves, can be interconnected by deformable valves, and can be included in or on themicrofluidic device500. According to various embodiments, adeformable valve536, for example, a Zbig valve, can be arranged between thefluid retainment regions514,516,518,520,522,524 to selectively control fluid communication between the fluidretain meat regions514,516,518,520,522,524. The deformable valve as described in U.S. patent application Ser. No. 10/808,228, filed Mar. 24, 2004, which is incorporated herein in its entirety by reference.
• According to various embodiments, thesubstrate510 of themicrofluidic device500 can be at least partially formed of a deformable material, for example, an inelastically deformable material. Thesubstrate510 can include a single layer of material, a coated layer of material, a multi-layered material, or a combination thereof. Thesubstrate510 can be formed as a single layer and made of a non-brittle plastic material, for example, polycarbonate, for example, a TOPAS material, a plastic cyclic olefin copolymer material available from Ticona (Celanese AG), Summit, N.J., USA. The thermal conductivity of the TOPAS cyclic olefin copolymer can be about 0.16 Watt per meter Kelvin. Thesubstrate510 can be in the shape of a disk, a rectangle, a square, or any other shape. Thesubstrate510 can provide an operative surface for a thermal device to thermally contact themicrofluidic device500.
• According to various embodiments, an elasticallydeformable cover sheet508 can be adhered to at least one of the surfaces of thesubstrate510. Thecover sheet508 can be made of, for example, a plastic, elastomeric, or other elastically deformable material. Themicrofluidic device500 can include a central axis of rotation534. Thecover sheet508 can provide an operative surface for a thermal device to thermally contact themicrofluidic device500. An input fluid-containment feature530 can be fluidly connected to a manifold532 for the introduction of one or more fluids to thefluid processing pathway512 via a branch channel (not shown) or opening a valve (not shown). For example, one or more fluids can be introduced by piercing through thecover sheet508 in the area of the input fluid-containment feature530 and injecting one or more fluids into the input fluid-containment feature530. The fluid can be collected in outputfluid retainment regions526,528 after being processed through thefluid processing pathway512. According to various embodiments, thefluid processing pathway512 can be arranged generally linearly. According to various embodiments, and as shown inFIG. 8 andFIG. 9, more than onefluid processing pathway512 can be arranged side-by-side in or on thesubstrate510. A plurality of samples or a plurality of reactions on the same sample can be processed in thefluid processing pathway512. The processing can be serial or simultaneous. For example, 12, 24, 48, 96, 192, or 384 of thefluid processing pathways512 can be arranged side-by-side to form a set of fluid processing pathways on a singlemicrofluidic device500. Moreover, two or more sets of fluid processing pathways can be arranged on a singlemicrofluidic device500. One or more outputfluid retainment regions526,528 can be provided for eachfluid processing pathway512.
• According to various embodiments, themicrofluidic device500 can be rotated through a central axis of rotation534, to selectively force fluids between thefluid retainment regions514,516,518,520,522,524 of themicrofluidic device500, by way of applying a centripetal force. For example, by spinning themicrofluidic device500 around the central axis of rotation534, a fluid can be selectively forced to move from at least the input fluid-containment feature530 to the outputfluid retainment regions526,528 along thefluid processing pathway512. The fluid flow can be controlled by manipulation ofvalves536. According to various embodiments, aplaten502, or a microfluidic device holder built-in theplaten502, can be arranged to support and rotate themicrofluidic device500 about the axis of rotation534 of the platen and/orholder502. According to various embodiments and as shown inFIG. 8, the axis of rotation of theplaten502 can be coaxial with the central axis of rotation534 of themicrofluidic device500. Analignment notch504 can be disposed in the microfluid.processing device500 to complement analignment pin506 in theplaten502.
• According to various embodiments, the ability to achieve disparate temperature zones in nearly abutting fluid retainment regions can allow adjacent fluid retainment regions of a fluid processing device to be loaded with reagents activated at disparate temperatures. Alternatively or additionally, temperature sensitive reagents can be disposed in a fluid retainment region while an adjacent fluid retainment region can be subjected to temperatures that would render the temperature sensitive reagents inoperative. According to various embodiments, the temperature sensitive reagents can maintain their effectiveness, even as the adjacent fluid retainment region is subjected to temperatures incompatible with the temperature sensitive reagents. Temperature sensitive reagents can be reagents that can lose their effectiveness or be rendered inoperable if subjected to a temperature above a lower limit. According to various embodiments, a fluid retainment region and an adjacent temperature sensitive reagent region can be disposed nearly abutting on a fluid processing device, for example, separated by from about 1 mm to about 50 mm, separated by about 10 mm, separated by about 3 mm, or separated by about 2 mm.
• According to various embodiments, the fluid movement device can provide for movement of fluids within features of a fluid processing device by, for example, spinning, suctioning, or pneumatics.
• According to various embodiments, the valve manipulation device can comprise, for example, a deformer, a flap, or a solenoid. Further information about valve manipulation devices can be found in U.S. patent application Ser. No. 10/808,229 filed Mar. 24, 2004, which is incorporated herein in its entirety.
• According to various embodiments,FIG. 9 is a top view of an exemplary microfluidic device800 having twoinput ports801,802 for distributing a fluid sample torespective flow distributors804,806, each flow distributor being in fluid communication with, or being designed to be in interruptible communication with apathway803. The microfluidic device800 can include 384output ports808. Eachpathway803 can include aPCR chamber814, aPCR purification chamber816, a flow restrictor, a vertical flow-splitter that leads to aforward sequencing chamber818 and areverse sequencing chamber820, a forward sequencingproduct purification chamber822, a reverse sequencingproduct purification chamber824, a purified forward sequencingproduct output chamber826, a purified reverse sequencingproduct output chamber828, a plurality of opening and closing valves, or a combination thereof. Thethermal device600 ofFIG. 3 can thermally regulate fluids disposed in the various chambers of the microfluidic device800. Thethermal device600 ofFIG. 3 can thermally regulate fluids disposed in thePCR chamber814 and thePCR purification chamber816 for eachpathway803. Thethermal device600 ofFIG. 3 can thermally regulate fluids disposed in theforward sequencing chamber818, thereverse sequencing chamber820, the forward sequencingproduct purification chamber822, and a reverse sequencingproduct purification chamber824, for eachpathway803. Exemplary volumes for each of the chambers can be from 0.01 micro liters to about 100 micro liters.
• According to various embodiments, the thermal device can be an assembly. The assembly can include several components that can be stacked or assembled together to form a single-side thermal device. The assembly can include, for example, a cold block, one or more thermoelectric devices, a first hot block, one or more thermoelectric devices, and another cold block. This embodiment of the thermal device can provide one hot region and two cold regions on fluid processing device, for example,thermal device140 ofFIG. 2. The thermal device assembly can be augmented by adding one or more thermoelectric devices, a first hot block, one or more thermoelectric devices, and another cold block to the thermal device as desired, for example,thermal device600 ofFIG. 3. A system can include a thermal device providing a plurality of hot blocks, for example, one, two or three hot blocks for the device ofFIG. 8 for heating, two, four or six hot blocks for the device ofFIG. 9. The hot blocks can correspond to a PCR chamber, a forward sequencing chamber and/or a reverse sequencing chamber of the microfluidic devices illustrated inFIG. 8 andFIG. 9, as desired. The hot blocks can be heated by operating adjacent thermoelectric devices, as desired.
• Thechart620 ofFIG. 10 is an example of a fluid being subjected to a thermal protocol such as PCR. Atemperature plot622 tracks the fluid temperature over time. The fluid was disposed in a fluid retainment region of a fluid processing device, for example,fluid retainment region121 ofFIG. 2, orfluid retainment region514 ofFIG. 8. A dual-sided thermal device, for example, the dual-sidedthermal system100 ofFIG. 2, was utilized to obtain the plot. The fluid retainment region was in thermal contact with two hot blocks, for example,hot blocks102 ofFIG. 2. Thefluid temperature plot622 cycles in the range from about 55° C. to about 95° C. Thetemperature plot624 depicts the temperature of a fluid in a second fluid retainment region, for example, a purification region. The second fluid retainment region, for example,fluid retainment region122 ofFIG. 2,fluid retainment region516 ofFIG. 8, can be in contact with two cold blocks, for example,cold blocks106 ofFIG. 2. The temperature of the second fluid retainment region can fluctuate in the range of from about 40° C. to about 45° C. According to various embodiments, the temperature measured for the fluid retainment region can fluctuate in a range of at least, for example, about 20° C., or about a 40° C. range, while the temperature of the second fluid retainment region fluctuates over a range of, for example, less than about 5° C., or less than about 10° C.
• Example.FIG. 11 is a chart of temperatures obtained using five thermocouples embedded or buried within a plastic card. The thermocouples were obtained from Omega Engineering. Data was acquired using LabView software from National Instruments of Austin, Tex. using their off-the shelf data acquisition PC board (see, www.ni.com/labview). A depression was machined in a surface of the substrate. The depression allowed for a flush placement, with respect the machined surface, of thermocouples and their respective signal wires in the substrate. A cover film was placed over the thermocouples. A single-sided thermal device including a hot block and a cold block was placed in thermal contact with the cover film. The thermal device was thermo-cycled by a control device to follow the PCR protocol. Data tracks designated Chan4, Chan6, and Chan8 tracked temperatures reported by some of the thermocouples, linearly arranged, in thermal contact with the hot block. Data tracks designated Chan5 and Chan7 tracked temperatures reported by some of the thermocouples, linearly arranged, in thermal contact with the cold block. The hot block was offset from the cold block by about 6 mm. A pressing device was used to sandwich the plastic card with the embedded thermocouples. A single-sided thermal device was disposed on one side of the plastic card. A device including a first aluminum block, a plastic hot block, a second aluminum block, and two fans directing air flow over the first and second aluminum blocks, was disposed on the other side or opposing surface of the plastic card.
• According to various embodiments, a fluid processing device can be used to manipulate a fluid, for example, a micro-sized fluid sample. The fluid sample can be subjected to at least two reactions in at least two respective reactions wells formed in the fluid processing device. The at least two reactions can transpire serially. A latter reaction within the set of the at least two reactions can use the product of the preceding reaction. The latter reaction well can be loaded with some reagents prior to activating a first reaction of the at least two reactions. According to various embodiments, to remain operative some of the reagents in the latter reaction well can require that the latter reaction well be maintained at a preservative temperature range exclusive and/or partially overlapping of thermal parameters of a reaction transpiring in the first well. According to various embodiments, a thermal device adapted to maintain the at least two reaction wells at different thermal parameters is described herein.
• According to various embodiments, a fluid processing device can be used to manipulate at least two fluid samples, for example, at least two micro-sized fluid samples. The at least two fluid samples can be subjected to at least two reactions in at least two respective reaction wells formed in the fluid processing device. The at least two reactions can require mutually exclusive and/or partially overlapping thermal parameters to transpire in parallel or at the same time.
• According to various embodiments, in a PCR device a PCR sample can be cycled in the temperature range of about 50° C. to about 95° C. A second fluid retainment region on the fluid processing device can concomitantly contain some size-exclusion ion-exchange (SEIE) beads. The SEIE beads can be rendered inoperative or less effective, when subjected to temperatures above about 50° C. Alternatively or additionally, the inadvertent heating of adjacent fluid retainment regions, for example, a second fluid retainment region, in the fluid processing device can expand any air contained therein. The air expansion can cause the adjacent fluid retainment regions and/or a seal disposed in the adjacent fluid retainment regions to burst or “blow-out.” Similar thermal parameters can be imposed by a sequencing device including a sequencing-fluid retainment region and a sequencing purification fluid retainment region containing SEIE beads. The thermal parameter requirements for a plurality of reactions can proliferate as an increasing number of reactions can transpire in a fluid processing device, for example, a fluid processing device including a PCR fluid retainment region, a PCR-purification fluid retainment region, a sequencing fluid retainment region, a sequencing purification fluid retainment region. The fluid processing device can additionally or alternatively include a forward sequencing fluid retainment region, a forward sequencing purification fluid retainment region, a reverse sequencing fluid retainment region, a reverse sequencing purification fluid retainment region. According to various embodiments, a fluid processing device can include a reaction region for an Oligonucleotide Ligation Assay isothermal biological assay.
• Other embodiments will be apparent to those skilled in the art from consideration of the present specification and practice of various embodiments disclosed herein. It is intended that the present specification and examples be considered as exemplary only with a true scope and spirit indicated by the teachings and equivalents thereof.

Claims (29)

1. A method comprising:

positioning a fluid processing device between and in thermal contact with a first thermal device on a first surface of the fluid processing device, and a second thermal device on an opposite second surface of the fluid processing device; and

regulating the temperature of the fluid processing device for carrying out a reaction by thermally cycling the first and second thermal devices such that each carries out a heating portion of a thermal cycle and a cooling portion of the thermal cycle.

2. The method ofclaim 1, wherein the fluid processing device comprises a plurality of fluid retainment regions, the plurality of fluid retainment regions comprising reaction components.

3. The method ofclaim 1, wherein the reaction carried out comprises an amplification reaction and the reaction components comprise a target nucleic acid material and a plurality of amplifying reagents.

4. The method ofclaim 1, further comprising holding the fluid processing device with a fluid processing device holder configured to position the fluid processing device in heat-transfer position with respect to the first thermal device and the second thermal device.

5. The method ofclaim 1, wherein the first thermal device and the second thermal device together comprise a dual-sided vertical Peltier thermal cycler.

6. The method ofclaim 1, wherein regulating the temperature of the plurality of fluid retainment regions comprises using a bi-directional power supply under computer control to affect the thermal cycling.

7. The method ofclaim 1, wherein each of the first thermal device and the second thermal device comprises at least one heat sink and the method further comprises directing air-flow over each of the heat sinks.

8. The method ofclaim 1, wherein the fluid processing device is compressible and the positioning comprises compressing the fluid processing device between the first thermal device and the second thermal device.

9. The method ofclaim 1, wherein the fluid processing device comprises a microfluidic card device.

10. The method ofclaim 1, wherein the first and second thermal devices are thermally cycled at the same time.

11. A system comprising:

a fluid processing device comprising a first surface and an opposite second surface;

a first thermal device;

a second thermal device; and

a fluid processing device holder for positioning the fluid processing device in a heat-transfer position between the first thermal device and the second thermal device, wherein the first surface is in thermal contact with the first thermal device, the second surface is in thermal contact with the second thermal device, and the first thermal device and the second thermal device are configured to operate such that each carries out a heating portion of a thermal cycle and a cooling portion of the thermal cycle.

12. The system ofclaim 11, wherein the fluid processing device contains a plurality of fluid retainment regions and the plurality of fluid retainment regions comprise reaction components.

13. The system ofclaim 12, wherein the reaction components comprise a target nucleic acid material and a plurality of amplifying reagents.

14. The system ofclaim 11, wherein each of the first thermal device and the second thermal device comprises a block, and each block comprises a flat surface.

15. The system ofclaim 11, wherein the first thermal device and the second thermal device together comprise a double-sided thermal cycler.

16. The system ofclaim 11, wherein the first thermal device and the second thermal device together comprise a dual-sided vertical Peltier thermal cycler.

17. The system ofclaim 11, wherein each of the first thermal device and the second thermal device comprises at least one heat sink.

18. The system ofclaim 11, comprising at least one fan adapted to create an air current in thermal contact with the first thermal device, the second thermal device, or both.

19. The system ofclaim 11, wherein the fluid processing device comprises a microfluidic card device.

20. The system ofclaim 11, wherein the fluid processing device is compressible.

21. The system ofclaim 11, wherein the plurality of amplifying reagents comprise polymerase chain reaction reagents.

22. The system ofclaim 11, wherein the first thermal device and second thermal device are configured to operate at the same time.

23. A system comprising:

a substrate comprising a first surface and an opposite second surface, and containing a plurality of fluid retainment regions;

a first thermal device;

a second thermal device; and

a substrate holder for positioning the substrate in a heat-transfer position between the first thermal device and the second thermal device, wherein the first surface is in thermal contact with the first thermal device, the second surface is in thermal contact with the second thermal device, and the first thermal device and the second thermal device are configured to operate such that each carries out a heating portion of a thermal cycle and a cooling portion of the thermal cycle.

24. The system ofclaim 23, wherein the plurality of fluid retainment regions comprise reaction components and the reaction components comprise a target nucleic acid material and a plurality of amplifying reagents.

25. The system ofclaim 23, wherein the substrate is formed at least partially from a deformable material.

26. The system ofclaim 23, wherein the substrate comprises a plastic material.

27. The system ofclaim 23, wherein each of the first thermal device and the second thermal device comprises a block, and each block comprises a flat surface.

28. The system ofclaim 23, wherein the first thermal device and the second thermal device together comprise a double-sided thermal cycler.

29. The system ofclaim 23, wherein the first thermal device and the second thermal device together comprise a dual-sided vertical Peltier thermal cycler.

Images