CN102513169A - 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 thereof Download PDFInfo
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- CN102513169A CN102513169A CN2011104078317A CN201110407831A CN102513169A CN 102513169 A CN102513169 A CN 102513169A CN 2011104078317 A CN2011104078317 A CN 2011104078317A CN 201110407831 A CN201110407831 A CN 201110407831A CN 102513169 A CN102513169 A CN 102513169A
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Abstract
The invention discloses a microfluidic device used in micron-grade particle high-flux separation, and a manufacturing method thereof. On the device, separating substrates and a functional substrate are sequentially stacked; a sample inlet communicates with a main separation runner; terminals of branched runners respectively communicate with separation outlets, sample outlets and the main separation runner; a sample outlet of each layer of separating substrate is stacked and sealed-connected with a sample inlet of a next-layer separating substrate. During a manufacturing process, micro runners on each layer of separating substrate are manufactured with a micro processing technology; through holes provided on each layer of separating substrate are adopted as particle inlets; stacking of the separating substrates are realized through aligning markers and a bonding technology; through holes are provided on bonded separating substrates, and are adopted as particle outlets; the functional substrate and the packed multiple layers of separating substrates are bonded and packaged. According to the micro-runner structure provided by the invention, the current velocity of an injected sample is improved, and a low Reynolds number concept of traditional microfluidic chips is broken through. High-flux and continuum dimension separation of micron-grade biological particles is realized with a micro-fluidic inertia effect.
Description
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 is realized fluid sample or micro-and nano-particles check and analysis, the new technology of function such as is controlled, synthesizes as a kind of micro-nano-scale; Because volume is little, cost is low and consume advantages such as the appearance amount is few, has been widely used in the detection and analytical applications of research fields such as clinical medicine, biochemical analysis, biology.Substitute expensive traditional cabinet type diagnostic analysis equipment, realize that manipulation function such as efficiently the transporting of micro Nano material, sorting, extraction, assembling and mixing have become important technology that enables in the micro-fluidic research.(Point-of-care testing, exploitation POCT) has the very important applying value of putting into practice and the efficient sorting that how to realize the minute yardstick object is for single cell analysis and clinical real-time test instrument.But present minute yardstick sorting technology by its mechanism brief overview be below several types: the first kind be from macroscopical filtering technique develop the micro-pore-film filtration technology of coming or based on intercept, the microscreening selecting technology of cross-flow structure, but problems such as such technology exists that versatility is poor, cost height and the easy obstruction of micro-structural; Second type of single game or many compound sorting technology that are based on electricity, sound, magnetic, light and external fluid, but generally there is outfield power consumption in such technology, is difficult for integrated microization and damages defective such as micro-nano biomaterial; The 3rd type of sorting or extractive technique that is based on complex micro structures such as micro-pillar array, wall V-type groove and reducing and expansion array, but still there are deficiencies such as processing technology complicacy, versatility difference in such technology.Except that problem and limitation that above-mentioned three major types technology exists, (Reynolds number is generally 10 owing to lower flow velocity in most micro-fluidic chips
-6～10
1) and the poor efficiency processing mode of mass, make its flux receive great restriction, can't satisfy that blood plasma extracts and the processing demands of bulk samples such as rare cell sorting.In addition, present most of micro-fluidic sorting chip only can be realized the separation of two kinds of sized particles.For overcoming this limitation, recent part Study is utilized the integrated order sorting that realizes many sized particles of plane many separation units function, but such technology has strengthened the size of chip largely, is unfavorable for the integrated microization of chip.
Summary of the invention
Goal of the invention: the deficiency that the objective of the invention is to overcome 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 the chip size prerequisite, realizing the continuous high-efficient sorting of multiple different size micron order biomone.
Technical scheme: the present invention realizes through following technical scheme; Device of the present invention comprises function substrate and two-layer at least sorting substrate; Said sorting substrate and function substrate sequential pile buttress; 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, and the end of fork runner is communicated with sorting outlet, sample export and main sorting runner respectively, and the sample export of every layer of sorting substrate piles up sealing-in with the sample inlet of following one deck sorting substrate.
Said 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 is communicated with the fork runner.
The cross section of said straight channel is the high-aspect-ratio rectangle, and the specific dimensions particle can balance be focused near the center, long limit (runner height).
The size relationship of the cross sectional dimensions of the straight channel of said every layer of sorting substrate and the maximum particle of this layer is:
a
p/ L
c>=0.07, wherein, a
pDiameter for the maximum particle of this layer; L
cCharacteristic size for straight channel.
Said every layer of sorting substrate is provided with alignment mark, convenient vertical alignment when making multilayer sorting substrate.
The material of said sorting substrate is selected from a kind of in dimethyl silicone polymer, Teflon, polymethyl methacrylate, SU-8 photoresist, glass, silicon and the quartz.
Said function substrate is slide or electrode.
Connection runner between said fork runner and the sample export is a polygon, can when substrate is thin, reduce the influence of levels fluid pressure to the substrate micro-structural.
The preparation method of the micro-fluidic device of a kind of micro-size particles high flux sorting may further comprise the steps:
(1) every layer of on-chip each fluid channel of sorting made through micro-processing technology, can realize through soft lithographic or other manufacturing process;
(2) get the inlet of through hole at the sample inlet place of every layer of sorting substrate as particle;
(3) pass through stacking and the encapsulation that alignment mark and bonding techniques are realized multilayer sorting substrate;
(4) the on-chip sorting outlet of each layer sorting is got the outlet of through hole as particle after encapsulating with upper strata sorting substrate or multilayer sorting substrate bonding;
(5) with the multilayer sorting substrate bonding encapsulation after function substrate and the encapsulation.
In the said step (4), the sorting of the superiors' sorting substrate outlet is got through hole before the bonding encapsulation, the sample export of bottom sorting substrate with top multilayer sorting substrate bonding after get the outlet of through hole as final sample.
Among the present invention; Relation between every layer of sorting substrate is to treat that sorting or purification sample are injected by the sample inlet of the superiors' sorting substrate with specific flow velocity through the external fluid driving arrangement and through the processing of the main sorting runner of the superiors' sorting substrate, focuses on by the effect of the microfluid inertia effect (inertia migration) particle that size is maximum and is separated to the fork runner and by sorting outlet derivation chip; The residual particles mixed liquor is imported to the sample inlet of second layer sorting substrate by the sample export of the superiors' sorting substrate; After the processing of the residual particles mixing suspension that imports through second layer sorting substrate master sorting runner; The maximum particle of size is still derived chip by the fork runner of second layer sorting substrate through the sorting outlet in the residual particles mixing suspension; Residual particles mixed liquor after second layer sorting substrate is handled is still continued to import the sample inlet of the 3rd layer of sorting substrate by the sample export of second layer sorting substrate; And the like until the processing of accomplishing all sorting substrate layer master sorting runners, derive final sample by the sample export of bottom sorting substrate at last.The sample inlet of each layer sorting substrate is accomplished punching before the bonding encapsulation; Except that inserting microtubule, the sample inlet place of the superiors' sorting substrate is used to connect the external fluid driving arrangement; The sample inlet of all the other each layer sorting substrates all piles up sealing-in with the sample export of last layer, is used to import the residual particles suspension that the sorting of sorting substrate master sorting runner is handled through the upper strata.Whether the outlet of each layer sorting substrate gets through the hole needs to choose according to its function; The purpose that is provided with like outlet is then need not punch to next sorting substrate output sample flow; As need directly with sample flow derive chip then need this layer sorting substrate and upper strata sorting substrate or above multilayer sorting substrate bonding encapsulation finishing back punching (punching before the bonding encapsulation of the superiors' substrate), and need other layer of assurance sorting substrate not have the micro-structural existence in this position.
Beneficial effect: the present invention compares prior art and has the following advantages; Fluid channel structure of the present invention is through 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 the micron order biomone, this technology only need be controlled the cross sectional dimensions and the runner overall length of runner, have simple in structure, need not, chip manufacturing low cost and other advantages high by outfield, flux; Efficient sorting when simultaneously, the present invention has realized multiple different size particle through 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 plane many separation units integrated technology, have advantages such as microminiaturization easy of integration; The detected object high flux sorting that device that the present invention proposes and preparation method can be widely used in fields such as clinical diagnosis, biological study, biochemical analysis, environmental monitoring.
Description of drawings
Fig. 1 is the vertical view of sorting substrate in the present invention;
Fig. 2 is the vertical 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 an A-A cross-sectional view among Fig. 3;
Fig. 5 is the vertical view behind the sorting substrate stacking about the present invention;
Fig. 6 is a fluid channel particle grading principle sketch map of the present invention.
The specific embodiment
Elaborate in the face of embodiments of the invention down, present embodiment provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment being to implement under the prerequisite with technical scheme of the present invention.
The sorting substrate material of present embodiment is dimethyl silicone polymer (PDMS), also can make with 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 material different, bonding techniques and finished product device are also inequality, should select suitable chip material according to actual processing and fabricating condition and application demand.PDMS micro-structural sorting substrate is made by soft lithography in the present embodiment, and cast used formpiston of sorting substrate adopts the processing of SU-8 maskless lithography, and this technology has flexible high, advantage such as cost of manufacture is low and the process-cycle is short.Also can be by making formpiston based on the technology such as etching processing of the chrome mask/photoetching technique of printing film mask, the wet method/deep reaction ion etching of silicon, ultraprecise machined, metal plating and photosensitive circuit plate; But characteristics such as its machining accuracy that can reach of different technologies and shape of cross section are also inequality, should select according to sorting object and physical device condition.
Present embodiment is double-deck micron order biomone high flux sorting device; Use soft lithography (the PDMS method of molding combines maskless lithography formpiston process technology) to make the two-layer PDMS sorting substrate that contains specific microstructure; Also can constitute in other embodiments by the sorting substrate more than three layers; Can under the indeformable situation of the contained micro-structural of assurance, reduce the thickness of single substrate, thereby realize the stacking of the maximum number of plies on the vertical space.The device of the said double-layer structure of present embodiment can be used for sorting or the micro particles of three kinds of different sizes of purifying, can be widely used in multiple micron order impurity in the extraction, municipal sewage of the separation that detects haemocyte in the blood sample and blood plasma removal and rare cell practical application such as separate.
As shown in Figure 1, the device of present embodiment comprises function substrate 3 and two-layer sorting substrate, last sorting substrate 1, sorting substrate 2 and function substrate 3 stacking in proper order on vertical space down.
As shown in Figure 2; Following sorting substrate 2 be provided with down main sorting runner, down diverge runner 12 and following sorting the outlet 13; Following sample inlet 11 and following sample export 14 are got by puncher; Straight channel 15 and following flaring runner 16 under main sorting runner comprises down in the present embodiment, following sample inlet 11 is communicated with following straight channel 15, the miner diameter end of following flaring runner 16 and following straight channel 15 connections; The end of runner 12 diverge down respectively with following sorting outlet 13, the bigger diameter end of sample export 14 and following flaring runner 16 is communicated with down, and the cross section of last straight channel 8 and following straight channel 15 is the high-aspect-ratio rectangle.Following sorting substrate 2 is provided with strip dihedral alignment mark 17, and the last sample export 7 of last sorting substrate 1 piles up sealing-in with the following sample inlet 11 of following sorting substrate 2.In the present embodiment; The connection runner that diverges down between runner 12 and the following sample export 14 is a rhombus; When avoiding individual layer sorting substrate thin; The sample gateway is to the influence of micro-structural on other substrates, and last sample inlet 4 can place the centre of diamond structure, and the motion of fluid in the fluid channel of sorting substrate 2 under when fluid pressure is big, also can not influencing.But when substrate layer was thicker, the drainage of only need punching did not have micro-structural with corresponding substrate place in addition, sample gateway and gets final product, and can arrange arbitrarily in other sample gateway.
Like Fig. 3 and shown in Figure 4, after two-layer PDMS micro-structural substrate preparation is accomplished, last sample inlet 4, go up sorting outlet 6, sorting outlet 13 down, sample inlet 11 needs before bonding encapsulates, get the individual layer through hole 18 of special pore size distribution down.After waiting to have beaten individual layer through hole 18 and two substrate micro-structural faces being cleaned up, present embodiment utilizes UV-irradiation oxygen to produce ozone, utilizes ozone that substrate surface is carried out the irreversible bonding that oxidation processes realizes two-layer sorting substrate.Also can carry out the irreversible bonding that surface modification realizes two-layer sorting substrate among other embodiment with means such as oxygen plasma treatment.Among other embodiment, also can when making substrate, allocate the proportioning of the different components (performed polymer and curing agent) of two sorting substrates, utilize the molecular diffusion effect on the interface to realize irreversible from bonding.During two-layer sorting stack of substrates buttress, by the auxiliary stacking of down realizing two-layer sorting substrate of the alignment mark on the sorting substrate 2 up and down at aligning equipments such as stereoscopes.The alignment mark of sorting substrate 2 is used about in the present embodiment, is positioned at the edge of substrate separately and is symmetrical distribution, can process with the contained fluid channel structure of sorting substrate through soft lithography.
But the Apparatus and method for that present embodiment proposed not is high especially for the requirement of alignment precision, as only going up the rough alignment between sorting outlet 6 and the following sample inlet 11 in the present embodiment.As shown in Figure 5, except that alignment mark, striped is filled to down the contained micro-structural of sorting substrate 2 among the figure, and black is filled to the contained micro-structural of sorting substrate 1.After two-layer sorting substrate bonding encapsulation finished, following sorting outlet 13 was all got double-deck through hole 19 with following sample export 14 places.Then, realize the sorting substrate of encapsulation back double-layer structure and the bonding of function substrate 3 with the uv/ozone process for modifying surface.The function substrate 3 of present embodiment is a slide.
After treating that the integral device encapsulation finishes, external fluid driving arrangements such as the individual layer through hole 18 insertion microtubules connection syringe pumps at last sample inlet 4 places are used for the specific flow velocity of sample to be introduced.Last sorting exports the individual layer through hole 18 at 6 places, and the double-deck through hole 19 at following sorting outlet 13 and following sample export 14 places inserts collection and the derivation that microtubule is used for sorting or purification back sample liquid respectively.Present embodiment has other particle branch of obvious difference in size with three kinds and elects job step and the principle that example is set forth this two-layer composite sorting device as.
As shown in Figure 6; Above sorting substrate 1 is an example; After injecting particle suspension with specific flow velocity in the runner; The particle of runner intake section random distribution is by descending macroparticle 20, middle particle 21 and the small-particle 22 of being divided into of particle diameter, and these three kinds of particles will receive fluid and pull the effect of power F1 and move along flow direction.Under the high flow velocities condition, because the parabolic type velocity profile 23 of uncle's howl leaf stream (Poiseuille flow) in the last straight channel 8, these three kinds of particles will receive the effect of the shear-induced inertia lift F2 that points to wall and move towards wall.When particle during near wall, owing to three kinds of particles because from then symmetrical tail that rotation produces is produced a wall that points to the runner center line by the wall influence induces inertia lift F3.
When particle size and channel size satisfy: a
p/ L
c>=0.07, wherein: a
pBe particle diameter; L
cCharacteristic size for straight channel; Can use minor face estimation for the straight channel of high-aspect-ratio rectangular cross section; Satisfying the particle of this condition will induce under the acting in conjunction of inertia lift F3 at above-mentioned shear-induced inertia lift F2 and wall; Based on the tubulose blockage effect, the particle that satisfies this condition partly focuses to equilbrium position L0 at runner exit, and other particle then still is randomly distributed in the runner; Main sorting runner on the sized according to macroparticle 20 in the present embodiment; Make macroparticle 20 satisfy above-mentioned condition and focus to equilbrium position L0, and middle particle 21 does not satisfy this condition with small-particle 22 still random dispersion is in runner, the flaring runner plays the effect of two equilbrium position distances drawing back particle.In the time of near macroparticle 20 moves to fork runner 5; Because wall induces inertia lift F3 to be cancelled; Macroparticle 20 will be from upwards sorting outlet 6 derivation of fork runner 5 under the independent effect of shear-induced inertia lift F2; Middle particle 21 and small-particle 22 are then from upwards sample export 7 derivation of last fork runner 5, thus the separation and the purifying of realization particle.Middle particle 21 gets into sorting substrate 2 down with small-particle 22, and following sorting substrate 2 contained main sorting runners down are designed to make remain in two kinds of particles and focus on than macroparticle inertia, and still are randomly distributed in the runner than small-particle.Therefore; After the effect of main sorting runner under two kinds of particle suspensions processes of residue; Derive chips than macroparticle by following sorting outlet 13, remaining smallest particles is by following sample export 14 derivation chips, from accomplishing the whole high purity separation process of three kinds of different size particles.The straight channel of the contained rectangular cross section of main sorting runner only needs rectangular cross section to meet the specified particle balance to focus on and the out-of-focus condition of other particle in the sorting substrate 2 up and down, and flow channel length satisfies the particle that makes all meet above-mentioned requirements and all can migrate to equilbrium position L0 and get final product.If the particle of more different sizes is arranged, can separate and purifying by the corresponding multilayer sorting substrate of corresponding setting.
The sandwich construction sorting device that proposes in the present embodiment does not need labyrinth auxiliary with the power consumption outfield; Have simple in structure, cost is low, the efficient advantages of higher, can be widely used in the Application Research such as environmental monitoring, clinical diagnosis treatment, biology and biochemical analysis.
Claims (10)
1. the micro-fluidic device of micro-size particles high flux sorting; It is characterized in that; Said device comprises function substrate and two-layer at least sorting substrate; Said sorting substrate and function substrate sequential pile buttress, every layer of sorting substrate is provided with sample inlet, main sorting runner, fork runner, sorting outlet and sample export, and 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, and the sample export of every layer of sorting substrate piles up sealing-in with the sample inlet of following one deck sorting substrate.
2. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting; It is characterized in that: said main sorting runner comprises straight channel and flaring runner; Sample inlet and straight channel are communicated with; The miner diameter end of flaring runner and straight channel are communicated with, and the bigger diameter end of flaring runner is communicated with the fork runner.
3. the micro-fluidic device of micro-size particles high flux according to claim 2 sorting is characterized in that: the cross section of said straight channel is the high-aspect-ratio rectangle.
4. the micro-fluidic device of micro-size particles high flux according to claim 2 sorting is characterized in that: the size relationship of the cross sectional dimensions of the straight channel of said every layer of sorting substrate and the maximum particle of this layer is:
a
p/ L
c>=0.07, wherein, a
pDiameter for the maximum particle of this layer; L
cCharacteristic size for straight channel.
5. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting is characterized in that: said every layer of sorting substrate is provided with alignment mark.
6. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting is characterized in that: said sorting substrate material is selected from a kind of in dimethyl silicone polymer, Teflon, polymethyl methacrylate, SU-8 photoresist, glass, silicon and the quartz.
7. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting is characterized in that: said function substrate is slide or electrode.
8. the micro-fluidic device of micro-size particles high flux according to claim 1 sorting is characterized in that: the connection runner between said fork runner and the sample export is a polygon.
9. the preparation method of the micro-fluidic device of micro-size particles high flux sorting is characterized in that, may further comprise the steps:
(1) every layer of on-chip each fluid channel of sorting made through micro-processing technology;
(2) get the inlet of through hole at the sample inlet place of every layer of sorting substrate as particle;
(3) pass through stacking and the encapsulation that alignment mark and bonding techniques are realized multilayer sorting substrate;
(4) the on-chip sorting outlet of each layer sorting is got the outlet of through hole as particle after encapsulating with upper strata sorting substrate or multilayer sorting substrate bonding;
(5) with the multilayer sorting substrate bonding encapsulation after function substrate and the encapsulation.
10. the preparation method of the micro-fluidic device of a kind of micro-size particles high flux according to claim 9 sorting; It is characterized in that; In the said step (4); The sorting of the superiors' sorting substrate outlet is got through hole before the bonding encapsulation, the sample export of bottom sorting substrate with top multilayer sorting substrate bonding after get the outlet of through hole as 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 |
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| 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 kind of micro-fluidic pipettor gun head |
| CN107159334A (en) * | 2017-06-16 | 2017-09-15 | 东南大学 | A kind of micro-fluidic pipettor gun head |
| CN108160126A (en) * | 2017-11-30 | 2018-06-15 | 东南大学 | Micro particles high throughput is enriched with micro-fluidic chip |
| 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 |
| CN112771364A (en) * | 2018-09-27 | 2021-05-07 | 京瓷株式会社 | Particle separation measuring device and particle separation measuring apparatus |
| CN112771364B (en) * | 2018-09-27 | 2024-04-26 | 京瓷株式会社 | Particle separation device and particle separation measurement device |
| CN110124757A (en) * | 2019-04-29 | 2019-08-16 | 南京师范大学 | A kind of micro particles sorting micro-fluidic device |
| CN110639627A (en) * | 2019-09-06 | 2020-01-03 | 东南大学 | Quickly assembled spliced microfluidic chip |
| CN111569964A (en) * | 2020-05-22 | 2020-08-25 | 苏州研材微纳科技有限公司 | Microfluidic device and preparation method thereof |
| 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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