CN102513169B - Microfluidic device used in micron-grade particle high-flux separation, and manufacturing method thereof - Google Patents
Microfluidic device used in micron-grade particle high-flux separation, and manufacturing method thereofDownload PDFInfo
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- CN102513169B CN102513169BCN201110407831.7ACN201110407831ACN102513169BCN 102513169 BCN102513169 BCN 102513169BCN 201110407831 ACN201110407831 ACN 201110407831ACN 102513169 BCN102513169 BCN 102513169B
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Abstract
Translated fromChineseDescription
Technical field
What the present invention relates to is a kind of micro-fluidic device based on the microfluid inertia effect and preparation method thereof, micro-fluidic device of in particular a kind of micro-size particles high flux sorting and preparation method thereof.
Background technology
Microflow control technique realizes as a kind of micro-nano-scale the new technology that fluid sample or micro-and nano-particles detect functions such as analyzing, control, synthesize, because volume is little, cost is low and consume the advantages such as sample amount is few, be widely used in the detection and analytical applications of the research fields such as clinical medicine, biochemical analysis, biology.Substitute the expensive cabinet type diagnostic analysis equipment of tradition, realizing the manipulation function such as efficiently the transporting of micro Nano material, sorting, extraction, assembling and mixing has become an important technology that enables in micro-fluidic research.And efficient separation how to realize micro scale object has the very important applying value of putting into practice for the exploitation of single cell analysis and clinical real-time test instrument (Point-of-care testing, POCT).Current minute yardstick sorting technology can brief overview be following a few class by its mechanism: the first kind be from macroscopical filtering technique develop the microporous membrane filter techniques that comes or based on intercepting, the microscreening selecting technology of cross-flow structure, but such technology exists that versatility is poor, cost is high and the problem such as the easy obstruction of micro-structural; Equations of The Second Kind is single game or many compound sorting technologies based on electricity, sound, magnetic, light and external fluid, but such technology generally exists outfield power consumption, is difficult for integrated micro and damages the defects such as micro-nano biomaterial; The 3rd class is sorting or the extractive technique based on complex micro structures such as micro-pillar array, wall V-type groove and reducing and expansion arrays, but such technology still exists the deficiencies such as processing technology is complicated, versatility is poor.Except problem and limitation that above-mentioned three major types technology exists, due to flow velocity lower in most micro-fluidic chips, (Reynolds number is generally 10-6～101) and the poor efficiency processing mode of mass, its flux is greatly limited, cannot meet that blood plasma extracts and the processing demands of the bulk samples such as rare cell sorting.In addition, most of micro-fluidic sorting chip only can be realized the separation of two kinds of sized particles at present.For overcoming this limitation, recent part Study is utilized the integrated order sorting that realizes many sized particles of many separation units of plane function, but such technology has strengthened the size of chip largely, is unfavorable for the integrated micro of chip.
Summary of the invention
Goal of the invention: the object of the invention is to overcome the deficiencies in the prior art, micro-fluidic device of a kind of micro-size particles high flux sorting and preparation method thereof is provided, do not increasing under chip size prerequisite, realizing the continuous high-efficient sorting of multiple different size micron order biomone.
Technical scheme: the present invention is achieved by the following technical solutions, device of the present invention comprises function substrate and at least two-layer sorting substrate, described sorting substrate and function substrate order stacking, every layer of sorting substrate is provided with sample inlet, main sorting runner, fork runner, sorting outlet and sample export, sample inlet and main sorting runner are communicated with, the end of fork runner is communicated with sorting outlet, sample export and main sorting runner respectively, the stacking sealing-in of sample inlet of the sample export of every layer of sorting substrate and lower one deck sorting substrate.
Described main sorting runner comprises straight channel and flaring runner, and sample inlet and straight channel are communicated with, and the miner diameter end of flaring runner and straight channel are communicated with, and the bigger diameter end of flaring runner and fork runner are communicated with.
The cross section of described straight channel is high-aspect-ratio rectangle, and specific dimensions particle can balance be focused near center, long limit (runner height).
The cross sectional dimensions of the straight channel of described every layer of sorting substrate and the size relationship of this layer of maximum particle are:
Ap/ Lc>=0.07, wherein, apdiameter for this layer of maximum particle; Lccharacteristic size for straight channel.
Described every layer of sorting substrate is provided with alignment mark, convenient vertical alignment when making multilayer sorting substrate.
The material of described sorting substrate is selected from a kind of in dimethyl silicone polymer, Teflon, polymethyl methacrylate, SU-8 photoresist, glass, silicon and quartz.
Described function substrate is slide or electrode.
Connecting passage between described fork runner and sample export is polygon, can when substrate is thinner, reduce the impact of levels fluid pressure on substrate micro-structural.
A preparation method for the micro-fluidic device of micro-size particles high flux sorting, comprises the following steps:
(1) every layer of on-chip each fluid channel of sorting made by micro-processing technology, can realize by soft lithographic or other manufacturing process;
(2) at the sample inlet place of every layer of sorting substrate, get through hole as the entrance of particle;
(3) by alignment mark and bonding techniques, realize stacking and the encapsulation of multilayer sorting substrate;
(4) the on-chip sorting outlet of each layer of sorting with upper strata sorting substrate or multilayer sorting substrate bonding packaging after get through hole as the outlet of particle;
(5) by the multilayer sorting substrate bonding packaging after function substrate and encapsulation.
In described step (4), through hole is got in the sorting of the superiors' sorting substrate outlet before bonding packaging, the sample export of bottom sorting substrate with the substrate of multilayer sorting above bonding after get through hole as the outlet of final sample.
In the present invention, pass between every layer of sorting substrate is to treat that sorting or purification sample are injected and pass through the processing of the main sorting runner of the superiors' sorting substrate by external fluid driving arrangement by the sample inlet of the superiors' sorting substrate with specific flow velocity, by the effect of the microfluid inertia effect (inertia migration), the particle of size maximum is focused on to be separated to and diverges runner and export derivation chip by sorting; Residual particles mixed liquor is imported to the sample inlet of second layer sorting substrate by the sample export of the superiors' sorting substrate; The residual particles mixing suspension importing is after the processing of the main sorting runner of second layer sorting substrate, in residual particles mixing suspension, the particle of size maximum is still exported and is derived chip by sorting by the fork runner of second layer sorting substrate, residual particles mixed liquor after second layer sorting substrate is processed is still continued to import the sample inlet of the 3rd layer of sorting substrate by the sample export of second layer sorting substrate, the like until complete the processing of the main sorting runner of all sorting substrate layers, finally by the sample export of bottom sorting substrate, derive final sample.The sample inlet of each layer of sorting substrate completes punching before bonding packaging, except inserting microtubule, the sample inlet place of the superiors' sorting substrate is used for connecting external fluid driving arrangement, the sample inlet of all the other each layer of sorting substrates all with the stacking sealing-in of sample export of last layer, for importing the residual particles suspension through the main sorting runner of upper strata sorting substrate sorting process.Whether each layer of sorting substrate outlet gets through hole need be chosen according to its function, as the object that arranges of outlet does not need punching for export sample stream to next sorting substrate, as need directly sample conductance is gone out chip need this layer of sorting substrate and upper strata sorting substrate or above punching (the superiors' substrate punches before bonding packaging) after multilayer sorting substrate bonding packaging, and need to guarantee that other layer of sorting substrate exists without micro-structural in this position.
Beneficial effect: the present invention has the following advantages compared to existing technology, fluid channel structure of the present invention is by suitably improving the flow velocity that injects sample, break through the idea of low reynolds number in traditional micro-fluidic chip, ingenious high flux, the Continuous Flow size separation that utilizes the microfluid inertia effect to realize micron order biomone, this technology only needs to control cross sectional dimensions and the runner overall length of runner, have simple in structure, without, chip manufacturing low cost and other advantages high by outfield, flux; Efficient separation when meanwhile, the present invention has realized multiple different size particle by multilayer separation unit stacking; Having overcome conventional sorting technology can only sorting or the limitation of the two kinds of different size particles of purifying; Compare with existing many separation units of plane integrated technology, there is the advantages such as microminiaturization easy of integration; The detected object high flux sorting that the device that the present invention proposes and preparation method can be widely used in the fields such as clinical diagnosis, biological study, biochemical analysis, environmental monitoring.
Accompanying drawing explanation
Fig. 1 is the top view of sorting substrate in the present invention;
Fig. 2 is the top view of sorting substrate under the present invention;
Fig. 3 is the stereogram that stacking provisions of the present invention and through hole distribute;
Fig. 4 is A-A cross-sectional view in Fig. 3;
Fig. 5 is the top view after the upper and lower sorting substrate of the present invention stacking;
Fig. 6 is fluid channel particle grading principle schematic diagram of the present invention.
The specific embodiment
Below embodiments of the invention are elaborated, the present embodiment is implemented take technical solution of the present invention under prerequisite, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
The sorting substrate material of the present embodiment is dimethyl silicone polymer (PDMS), also can make of the polymer such as Teflon, polymetylmethacrylate, SU-8 photoresist, glass, silicon or quartz and hard material.Optics, the electrology characteristic of its corresponding processing technology of different materials, bonding techniques and finished product device are not identical yet, should select suitable chip material according to actual processing and fabricating condition and application demand.In the present embodiment, PDMS micro-structural sorting substrate is made by soft lithography, and cast formpiston used of sorting substrate adopts the processing of SU-8 maskless lithography, and this technology has flexible high, the advantage such as cost of manufacture is low and the process-cycle is short.Also can make formpiston by the technology such as lithography of wet method/deep reaction ion etching, ultraprecise machined, metal plating and the photosensitive circuit plate of the photoetching technique based on chrome mask/printing film mask, silicon, but the characteristics such as its machining accuracy that can reach of different technologies and shape of cross section are not identical yet, should select according to sorting object and physical device condition.
The present embodiment is double-deck micron order biomone high flux sorting device, use soft lithography (PDMS method of molding is in conjunction with maskless lithography formpiston process technology) to make the two-layer PDMS sorting substrate containing specific microstructure, also can be formed by three layers of above sorting substrate in other embodiments, can guarantee to reduce the thickness of single substrate in the indeformable situation of contained micro-structural, thereby realize the stacking of the maximum number of plies on vertical space.Described in the present embodiment, the device of double-layer structure can be used for sorting or the micro particles of the three kinds of different sizes of purifying, and can be widely used in the practical application such as separated that detects the removal of multiple micron order impurity in the separation of haemocyte and the extraction of blood plasma in blood sample, municipal sewage and rare cell.
As shown in Figure 1, the device of the present embodiment comprises function substrate 3 and two-layer sorting substrate, and upper sorting substrate 1,lower sorting substrate 2 and function substrate 3 be order stacking on vertical space.
Upper sorting substrate 1 is provided with main sorting runner,upper fork runner 5 andupper sorting outlet 6,upper sample inlet 4 andupper sample export 7 are got by puncher, in the present embodiment, upper main sorting runner comprisesstraight channel 8 andupper flaring runner 9,upper sample inlet 4 and upperstraight channel 8 are communicated with, the miner diameter end ofupper flaring runner 9 and upperstraight channel 8 are communicated with, the end ofupper fork runner 5 is communicated with the bigger diameter end ofupper sorting outlet 6,upper sample export 7 andupper flaring runner 9 respectively, and upper sorting substrate 1 is provided with FilledRectangle alignment mark 10.
As shown in Figure 2,lower sorting substrate 2 is provided with lower main sorting runner,lower fork runner 12 andlower sorting outlet 13,lower sample inlet 11 andlower sample export 14 are got by puncher, in the present embodiment, lower main sorting runner comprises lowerstraight channel 15 andlower flaring runner 16,lower sample inlet 11 and lowerstraight channel 15 are communicated with, the miner diameter end oflower flaring runner 16 and lowerstraight channel 15 are communicated with, the end oflower fork runner 12exports 13 with lower sorting respectively, the bigger diameter end oflower sample export 14 andlower flaring runner 16 is communicated with, the cross section of upperstraight channel 8 and lowerstraight channel 15 is high-aspect-ratio rectangle.Lower sorting substrate 2 is provided with stripdihedral alignment mark 17, theupper sample export 7 of upper sorting substrate 1 and the stacking sealing-in oflower sample inlet 11 of lower sorting substrate 2.In the present embodiment, connecting passage betweenlower fork runner 12 andlower sample export 14 is rhombus, when avoiding individual layer sorting substrate thinner, the impact of sample gateway on micro-structural on other substrates,upper sample inlet 4 can be placed in the centre of diamond structure, and the motion of fluid in the fluid channel ofsorting substrate 2 under also can not affecting when fluid pressure is larger.But when substrate layer is thicker, the drainage that only needs to punch is with corresponding other substrate place, sample gateway without micro-structural, can arrange arbitrarily in other sample gateway.
As shown in Figure 3 and Figure 4, after prepared by two-layer PDMS micro-structural substrate,upper sample inlet 4,upper sorting outlet 6,lower sorting outlet 13,lower sample inlet 11 need be got the individual layer through hole 18 of special pore size distribution before bonding packaging.After having beaten individual layer through hole 18 and two substrate micro-structural faces are cleaned up, the present embodiment utilizes UV-irradiation oxygen to produce ozone, utilizes ozone to carry out to substrate surface the irreversible bonding that oxidation processes realizes two-layer sorting substrate.In other embodiment, also can carry out the irreversible bonding that surface modification realizes two-layer sorting substrate by means such as oxygen plasma treatment.In other embodiment, the proportioning that also can allocate the different components (performed polymer and curing agent) of two sorting substrates when making substrate, utilizes the molecular diffusion effect on interface to realize irreversible from bonding.During two-layer sorting stack of substrates buttress, by the alignment mark on upper andlower sorting substrate 2, under the aligning equipments such as stereoscope auxiliary, realize the stacking of two-layer sorting substrate.In the present embodiment, the alignment mark of upper andlower sorting substrate 2 is used in conjunction with, and is positioned at separately the edge of substrate and is symmetric, and can together with the soft lithography fluid channel structure contained with sorting substrate, process.
But the Apparatus and method for that the present embodiment proposes not is high especially for the requirement of alignment precision, as only needed the rough alignment betweenupper sorting outlet 6 andlower sample inlet 11 in the present embodiment.As shown in Figure 5, in figure, except alignment mark, striped is filled to the contained micro-structural oflower sorting substrate 2, and black is filled to the contained micro-structural of sorting substrate 1.After two-layer sorting substrate bonding packaging, double-deck through hole 19 is all got atlower sorting outlet 13 andlower sample export 14 places.Then, with UV/ozone process for modifying surface, realize the rear sorting substrate of double-layer structure of encapsulation and the bonding of function substrate 3.The function substrate 3 of the present embodiment is slide.
After treating integral device encapsulation, the individual layer through hole 18 atupper sample inlet 4 places inserts microtubule and connects the external fluid driving arrangements such as syringe pump for the specific flow velocity introducing of sample.Upper sorting exports the individual layer through hole 18 at 6 places, and the double-deck through hole 19 atlower sorting outlet 13 andlower sample export 14 places inserts respectively microtubule for collection and the derivation of sorting or the rear sample liquid of purifying.The present embodiment has other particle of obvious difference in size with three kinds and divides job step and the principle of electing this two-layer composite sorting device of example elaboration as.
As shown in Figure 6, above sorting substrate 1 is example, with specific flow velocity, in runner, inject after particle suspension, the particle of runner intake section random distribution is by descendingmacroparticle 20,middle particle 21 and the small-particle 22 of being divided into of particle diameter, and these three kinds of particles pull the effect of power F1 and move along flow direction being subject to fluid.Under high flow velocities condition, due to the parabolictype velocity profile 23 of uncle's howl leaf stream (Poiseuille flow) in upperstraight channel 8, these three kinds of particles by be subject to pointing to wall shear-induced inertia lift F2 effect and towards wall, move.When particle is during near wall, due to three kinds of particles because from then symmetrical tail that rotation produces by wall, affected and produce a wall induction inertia lift F3 who points to runner center line.
When particle size and channel size meet: ap/ Lc>=0.07, wherein: apfor particle diameter, Lccharacteristic size for straight channel, can be with minor face estimation for the straight channel of high-aspect-ratio rectangular cross section, meet the particle of this condition by under the acting in conjunction at above-mentioned shear-induced inertia lift F2 and wall induction inertia lift F3, based on tubulose blockage effect, the particle that meets this condition partly focuses to equilbrium position L0 at runner exit, other particle is still randomly distributed in runner, in the present embodiment, according to the size ofmacroparticle 20, make upper main sorting runner, makemacroparticle 20 meet above-mentioned condition and focus to equilbrium position L0, andmiddle particle 21 and small-particle 22 do not meet this condition still random dispersion in runner, flaring runner plays the effect of two equilbrium position distances pulling open particle.In the time ofnear macroparticle 20 moves toupper fork runner 5, due to wall induction inertia lift, F3 is cancelled,macroparticle 20 is by upwards sortingoutlet 6 derivation ofrunner 5 of diverging from above under the independent role at shear-induced inertia lift F2,middle particle 21 and small-particle 22 fromupper fork runner 5 upwardssample export 7 derive, thereby realize separation and the purifying of particle.Middle particle 21 and small-particle 22 enterlower sorting substrate 2, and the contained lower main sorting runner design oflower sorting substrate 2 focuses on compared with macroparticle inertia for making to remain in two kinds of particles, and is still randomly distributed in runner compared with small-particle.Therefore, when two kinds of particle suspensions of residue are after the effect of lower main sorting runner, compared with macroparticle, bylower sorting outlet 13, derive chip, remaining smallest particles derives chip bylower sample export 14, from completing the whole high purity separation process of three kinds of different size particles.In upper andlower sorting substrate 2, the straight channel of the contained rectangular cross section of main sorting runner only needs rectangular cross section to meet specified particle balance to focus on and the out-of-focus condition of other particle, and flow channel length meets and makes all particles that meet above-mentioned requirements all can migrate to equilbrium position L0.If there is the particle of more different sizes, corresponding multilayer sorting substrate can be set accordingly and carries out separation and purifying.
The sandwich construction sorting device proposing in the present embodiment does not need labyrinth and power consumption outfield to assist, have simple in structure, cost is low, efficiency advantages of higher, can be widely used in the research such as environmental monitoring, clinical diagnosis treatment, biology and biochemical analysis application.
Claims (9)
1. the micro-fluidic device of micro-size particles high flux sorting, it is characterized in that, described device comprises function substrate and at least two-layer sorting substrate, described sorting substrate and function substrate order stacking, every layer of sorting substrate is provided with sample inlet, main sorting runner, fork runner, sorting outlet and sample export, sample inlet and main sorting runner are communicated with, the end of fork runner is communicated with sorting outlet, sample export and main sorting runner respectively, the stacking sealing-in of sample inlet of the sample export of every layer of sorting substrate and lower one deck sorting substrate; Described main sorting runner comprises straight channel and flaring runner, and sample inlet and straight channel are communicated with, and the miner diameter end of flaring runner and straight channel are communicated with, and the bigger diameter end of flaring runner and fork runner are communicated with.
2. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: the cross section of described straight channel is high-aspect-ratio rectangle.
3. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: the cross sectional dimensions of the straight channel of described every layer of sorting substrate and the size relationship of this layer of maximum particle are:
Ap/ Lc>=0.07, wherein, apdiameter for this layer of maximum particle; Lccharacteristic size for straight channel.
4. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: described every layer of sorting substrate is provided with alignment mark.
5. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: described sorting substrate material is selected from a kind of in dimethyl silicone polymer, Teflon, polymethyl methacrylate, SU-8 photoresist, glass, silicon and quartz.
6. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: described function substrate is slide or electrode.
7. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting, is characterized in that: the connecting passage between described fork runner and sample export is polygon.
8. a preparation method for the micro-fluidic device of micro-size particles high flux sorting, is characterized in that, comprises the following steps:
(1) every layer of on-chip each fluid channel of sorting made by micro-processing technology;
(2) at the sample inlet place of every layer of sorting substrate, get through hole as the entrance of particle;
(3) by alignment mark and bonding techniques, realize stacking and the encapsulation of multilayer sorting substrate;
(4) the on-chip sorting outlet of each layer of sorting with upper strata sorting substrate or multilayer sorting substrate bonding packaging after get through hole as the outlet of particle;
(5) by the multilayer sorting substrate bonding packaging after function substrate and encapsulation.
9. the preparation method of the micro-fluidic device of a kind of micro-size particles high flux according to claim 8 sorting, it is characterized in that, in described step (4), through hole is got in the sorting outlet of the superiors' sorting substrate before bonding packaging, the sample export of bottom sorting substrate with the substrate of multilayer sorting above bonding after get through hole as the outlet of final sample.
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| CN105344390B (en)* | 2015-11-20 | 2017-11-21 | 深圳市星国华先进装备科技有限公司 | A kind of alignment methods of micro-fluidic chip |
| CN106994369A (en)* | 2017-05-22 | 2017-08-01 | 东南大学 | Regulatable micro-fluidic integrated device of flux and preparation method thereof |
| CN107159334B (en)* | 2017-06-16 | 2019-04-30 | 东南大学 | A microfluidic pipette tip |
| CN108160126B (en)* | 2017-11-30 | 2020-05-19 | 东南大学 | Micro-fluidic chip for high-throughput enrichment of micro-particles |
| CN108279192A (en)* | 2018-01-29 | 2018-07-13 | 北京市劳动保护科学研究所 | A kind of pellet grain diameter grading device |
| JP6984037B2 (en)* | 2018-09-27 | 2021-12-17 | 京セラ株式会社 | Particle separation measurement device and particle separation measurement device |
| CN110124757B (en)* | 2019-04-29 | 2021-02-19 | 南京师范大学 | A micro-particle sorting microfluidic device |
| CN110639627A (en)* | 2019-09-06 | 2020-01-03 | 东南大学 | A Rapidly Assembled Spliced Microfluidic Chip |
| CN111569964B (en)* | 2020-05-22 | 2022-03-04 | 苏州研材微纳科技有限公司 | Microfluidic device and preparation method thereof |
| CN113176631A (en)* | 2021-05-11 | 2021-07-27 | 嘉兴微智光子科技有限公司 | Manufacturing method of optical waveguide and photonic device structure based on fluid channel |
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