CN105536895A - Openable micro-fluidic chip and preparation method thereof - Google Patents
Openable micro-fluidic chip and preparation method thereof Download PDFInfo
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- CN105536895A CN105536895A CN201510915541.1A CN201510915541A CN105536895A CN 105536895 A CN105536895 A CN 105536895A CN 201510915541 A CN201510915541 A CN 201510915541A CN 105536895 A CN105536895 A CN 105536895A
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- substrate
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- fluidic chip
- lug boss
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
Abstract
The invention discloses an openable micro-fluidic chip. The openable micro-fluidic chip comprises a first substrate and a second substrate, wherein the first substrate is provided with a recessed part, the second substrate is provided with a protruding part, the protruding part of the second substrate covers the surface of the recessed part of the first substrate, the periphery of the second substrate is bonded with the first substrate through a polymer, and the polymer is used for reversible connection with the first substrate and the second substrate, thereby forming an openable micro-fluidic structure. The invention further discloses a preparation method for the openable micro-fluidic chip. The openable micro-fluidic chip provided by the invention can be used for capturing cells or particulates and collecting the captured cells or particulates, so subsequent analysis is facilitated.
Description
Technical field
The invention belongs to micro-fluidic chip field, more specifically, relate to a kind of openable micro-fluidic chip and preparation method thereof.
Background technology
Micro-fluidic chip is the core of micro-total analysis system, the invention of micro-fluidic chip impels the analytical instrument being applied to chemistry, food, environment, medical science and life science to microminiaturization, automation, integrated and portability future development, its advantage is to detect various chemical reaction by micro-fluidic chip, remarkable reduction reagent consumes, and substantially increase analysis efficiency, reduce expense.Design and produce and can be applied to different analysis fields with the micro-fluidic chip of microfluidic channel network composition, comprise the multiple fields such as chemical analysis, food hygiene, environmental monitoring, medical science chemical examination, life science, Forensic science and national defence.
At present in micro-fluidic chip, utilize photoelectric technology or mechanically actuated to manipulate cell and microparticle more.The instrument that optical means relates to is complicated, manipulates single, limits more.Electrical method often needs integrated fine electrode on chip, needs complicated Energy control, and voltage on cell has adverse influence simultaneously.Mechanically actuated then lacks flexibility and the invertibity of operation; Such as, conventional cell mechanical positioning element is as micro-dam and micropore because its adjusting height is fixed, and cell, once captured, is difficult to remove completely, also there is certain difficulty to catching next step research after discharging simultaneously.Simultaneously, micro-fluidic chip of the prior art is formed with after two panels substrate bonding usually, substrate combines very tight, even if capture cell and particulate to be also difficult to open, to obtain required cell and particulate, or open reluctantly and can not ensure to catch accurately, being difficult to fix a point or required cell or particulate in directed acquisition micro-fluidic chip.
Summary of the invention
For above defect or the Improvement requirement of prior art, the invention provides a kind of openable micro-fluidic chip, its object is to connection first substrate by reversible polymer and the second substrate, thus form openable micro-fluidic structure, thus directed obtain needed for cell or particulate.
For achieving the above object, according to one aspect of the present invention, provide a kind of openable micro-fluidic chip, comprise the first substrate and the second substrate, described first substrate has depressed part, and described second substrate has lug boss, the lug boss of described second substrate is covered in the depressed part surface of described first substrate, and surrounding is combined with described first substrate by polymer, described polymer is used for reversible connection first substrate and the second substrate, thus forms openable micro-fluidic structure.
Preferably, the width of the lug boss of described second substrate and the lug boss junction of described first substrate is 1mm ~ 2mm.
Preferably, the height of the lug boss of described second substrate is greater than 20 μm.
Preferably, the depressed part of described first substrate is rectangle, and described depressed part reduces gradually in the length direction degree of depth of rectangle, and described micro-fluidic chip is used for catching of cell or particulate.
As further preferably, the degree of depth of the depressed part of described first substrate is 1 μm ~ 200 μm.
As further preferably, the length of the depressed part of described first substrate is 50mm ~ 70mm, and width is 10mm ~ 22mm.
As further preferably, described micro-fluidic chip is provided with entrance and exit, it is connected with the bosom of the depressed part of described first substrate and the most shallow place respectively.
As further preferably, described entrance and exit is arranged on the first substrate.
As further preferably, the surface coverage of the lug boss of described second substrate has the gel layer of 1 μm ~ 5 μm.
As still more preferably, described gel layer is agarose, shitosan or polyvinyl alcohol.
Preferably, described first substrate and the second substrate are glass.
Preferably, described polymer is dimethyl silicone polymer or epoxy resin.
According to another aspect of the present invention, provide a kind of preparation method of above-mentioned micro-fluidic chip, comprise the following steps:
(1) depressed part is obtained in the first substrate surface etching, lug boss is obtained in the second substrate surface etching, make the lug boss of described second substrate identical with the shape of the depressed part of described first substrate, the profile 1mm ~ 2mm larger than the profile of the depressed part of described first substrate of the lug boss of described second substrate;
(2) lug boss of described second substrate aimed at the depressed part of the first substrate fit, make the lug boss of described second substrate cover the depressed part of described first substrate completely, and with the fitting of described first substrate;
(3) surrounding to the lug boss of described second substrate fills prefabricated polymer, the lug boss of described second substrate is combined with described first substrate, namely obtains described micro-fluidic chip after described polymer cure.
Preferably, between described step (1) and described step (2), also comprise, at the gel layer of the lug boss surface coverage 1 μm ~ 5 μm of described second substrate.
In general, the above technical scheme conceived by the present invention compared with prior art, owing to passing through connection first substrate and second substrate of reversible polymer, can obtain following beneficial effect:
1, be bulge-structure by the second substrate designs, be covered in the surface of the first substrate depressed part, utilize polymer-filled in the circumference of the second substrate lug boss simultaneously, make the combination that the first substrate and the second substrate can be reversible, thus convenient micro-fluidic chip of opening is to obtain cell wherein or particulate;
2, preferably depressed part is set to the rectangle of degree of depth inequality, thus by the cell of different size or particle capture in the different parts of depressed part;
3, preferably using the material of glass as the first substrate and the second substrate, because glass is transparent material, the acquisition procedure of cell and particulate is more controlled;
4, the preparation method of this micro-fluidic chip is simple, and efficiency is high, is convenient to required cell and particulate in directed acquisition micro-fluidic chip.
Accompanying drawing explanation
Fig. 1 is the first substrate structure schematic diagram in embodiment 1;
Fig. 2 is the second substrate structure schematic diagram in embodiment 1;
Fig. 3 is microfluidic chip structure schematic diagram in embodiment 1;
Fig. 4 is that the micro-fluidic chip utilizing embodiment 1 to prepare catches cell schematic diagram;
Fig. 5 is that in the micro-fluidic chip of embodiment 1 preparation, cell schematic diagram is caught in paraffin encapsulation;
Fig. 6 is that micro-fluidic chip prepared by embodiment 1 opens rear transitional cell schematic diagram;
In all of the figs, identical Reference numeral is used for representing identical element or structure, wherein: 1-first substrate, 2-second substrate, 3-chip entrance, 4-chip exports, 5-first substrate depressed part, 6-second substrate lug boss, 7-PDMS, 8-raceway groove bosom, the most shallow place of 9-raceway groove, 11-marks, 12-cell.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.In addition, if below in described each embodiment of the present invention involved technical characteristic do not form conflict each other and just can mutually combine.
The invention provides a kind of openable micro-fluidic chip, comprise the first substrate and the second substrate, described first substrate has depressed part, described second substrate has lug boss, the lug boss of described second substrate is covered in the depressed part surface of described first substrate, and surrounding is combined with described first substrate by polymer, the width of its junction is 1mm ~ 2mm; Described polymer is dimethyl silicone polymer or epoxy resin, for reversible connection first substrate and the second substrate, thus forms openable micro-fluidic structure.Described first substrate and the second substrate are preferably glass, make the acquisition procedure of cell and particulate more controlled.The height of the lug boss of the second substrate is greater than 20 μm, makes the binding site of the second substrate and the first substrate have certain space, to utilize instrument to be separated the first substrate and the second substrate when opening.
Micro-fluidic chip for catching cell and particulate can utilize this design.Chip carry out cell and particulate catch experiment after, opening chip, thus cell needed for obtaining or particulate.
Such as, the depressed part of the first substrate can be designed to length is 50mm ~ 70mm, width is the rectangular configuration of 10mm ~ 22mm, the degree of depth of described rectangular configuration is 1 μm ~ 200 μm, its degree of depth reduces gradually at length direction, make the cell of different-diameter and particulate can be fixed on the diverse location of rectangle, micro-fluidic chip is provided with entrance and exit, it is connected with the bosom of depressed part and the most shallow place respectively.The size of the cell that its concrete degree of depth can be caught as required or particulate designs, such as, when needs are caught and are mixed with Normocellular circulating tumor cell, the most shallow for the degree of depth place can be designed to 5 μm, bosom is designed to 60 μm, thus the diverse location in rectangle length direction, catch and obtain 15 μm ~ 30 μm and the cell differed in size.Can in lug boss surface coverage as agarose, shitosan, the gel layer of the thickness 1 μm ~ 5 μm that the materials such as polyvinyl alcohol are formed, in the process that chip is used, cell or particulate better adhere to one side of lug boss, when being convenient to the unlatching of further microfluidic control chip, carrying out transfer and are separated.
Make use of after chip carries out catching experiment, cell or particulate can adhere to the side of the second substrate lug boss usually, and now the concrete open method of this micro-fluidic chip is as follows:
(1) alcohol is passed into from porch by emptying for the gas (bubble) micro-fluidic chip;
(2) from porch phosphate buffer solution PBS (formula: take NaCl8g, KCl0.2g, Na
2hPO412H
2o3.63g, KH
2pO40.24g, be dissolved in 900ml distilled water, with hydrochloric acid adjust pH to 7.4, add water and be settled to 1L, normal temperature saves backup) by emptying for the alcohol in micro-fluidic chip;
(3) pass into paraffin from porch, make paraffin be full of micro-fluidic chip;
(4) after paraffin solidification, the polymeric layer of the second substrate lug boss surrounding is removed with instruments such as blades, be separated the first substrate and described second substrate lug boss peripheral region, chip can be opened, obtain being encapsulated in the cell in paraffin or particulate, be separated by paraffin containing required cell or particulate, heating makes paraffin dissolve again, and can take out required cell or particulate to carry out next step research.
The preparation method of this micro-fluidic chip, comprises the following steps:
(1) depressed part is obtained in the first substrate surface etching, lug boss is obtained in the second substrate surface etching, make the lug boss of described second substrate identical with the shape of the depressed part of described first substrate, the profile 1mm ~ 2mm larger than the profile of the depressed part of described first substrate of the lug boss of described second substrate;
(2) lug boss of described second substrate aimed at the depressed part of the first substrate fit, make the lug boss of described second substrate cover the depressed part of described first substrate completely, and with the fitting of described first substrate;
(3) surrounding to the lug boss of described second substrate fills prefabricated polymer (being generally the mixture of polymer monomer and crosslinking agent), the lug boss of described second substrate is combined with described first substrate, after described polymer cure, namely obtains described micro-fluidic chip.
Embodiment one can open cavity heights controllable precise cell one side transfer micro flow chip
Step 1 prepares the first substrate
Select long 75mm, the slide of wide 25mm is as the material of the first substrate 1, raceway groove 5 is designed to long 60mm, wide 15mm, its degree of depth reduces from left to right gradually, bosom, left side is 60 μm, and the most shallow place, right side is 5 μm, uses the method for wet etching to carry out etching with the speed of 1mm/min and obtains corresponding pattern.And obtaining entrance 3 and outlet 4 at the central finishing impression at bosom and the most shallow place respectively, the side view of the first substrate obtained and top view are respectively as illustrated in figs. ia and ib.
Step 2. prepares the second substrate
Second substrate 2 is designed to 61mm × 16mm, is highly about the boss 6 of 80 μm, with and the similar method of the first substrate prepare the second substrate, its side view and top view are distinguished as shown in Figure 2 a and 2 b.
Step 3. configures the agarose solution of 2% mass fraction
Under the effect of magnetic stirring apparatus, solution mixes by 60 degree of temperature, then drip on boss with the solution that the pipettor of 1000 μ l gets 500 μ l, take advantage of agarose also uncolled agglutination time put into sol evenning machine, with 800rpm20s, the parameter of 1400rpm40s is got rid of even, finally in thermal station, dries to obtain the agarose film of about 2 μm with 50 DEG C.
Step 4. chip is fitted
The first substrate made is aimed at membrane second substrate, makes boss cover the first on-chip raceway groove completely, stack and make together with the first substrate fits tightly with the second substrate together.
Step 5. polymer cure
Select prefabricated PDMS (quality of polymer monomer and crosslinking agent is 10:1), be embedded into after around boss until PDMS, be heated to 85 DEG C within 10 minutes, make PDMS crosslinking curing after chip namely complete, the chip side view obtained and top view are respectively as shown in Figure 3 a and Figure 3 b shows, its raceway groove bosom 8 is 60 μm, and the most shallow place 9 of raceway groove is 5 μm.
Can carry out the test of controllable cell one side transfer micro flow chip with this micro-fluidic chip, its specific experiment process is as follows:
(1) in chip, alcohol is passed into by emptying for the gas (bubble) in micro-fluidic chip with the speed of 100 μm/min;
(2) in chip, phosphate buffer solution PBS (formula: take NaCl8g, KCl0.2g, Na is passed into the speed of 100 μm/min
2hPO412H
2o3.63g, KH
2pO40.24g, be dissolved in 900ml distilled water, with hydrochloric acid adjust pH to 7.4, add water and be settled to 1L, normal temperature saves backup) by emptying for the alcohol in raceway groove;
(3) in chip, concentration is passed into for 10 with the speed of 100 μm/min
3/ ml and diameter at 10 μm ~ 20 μm and the target cell differed in size, keep 2 minutes;
(4) in chip, pass into phosphate buffer solution PBS with the speed of 100 μm/min, make cell stay in described micro-fluidic chip different depth place according to the size of self;
(5) examine under a microscope the position of cell 11 stop and do mark 12, as shown in Figure 4;
(6) in chip, paraffin is passed into the speed of 100 μm/min;
(7) treat that paraffin solidifies, then with several positions that microscopic examination is marked, at this moment can see that the position of cellular retention and quantity do not change, as shown in Figure 5;
(8) at this moment, again chip is put in the water of 47 degrees Celsius and soak, the gel rubber material of bottom is softened, the PDMS around boss is removed with specific purpose tool again after ten minutes, open chip and just can obtain the paraffin section being bonded at boss face, arrive several positions that basis of microscopic observation is marked again, at this moment can see that the position of cellular retention and quantity still do not change;
(9) can obtain after paraffin being opened being encapsulated in the cell in paraffin layer or particulate, take out the paraffin of required cell place part again, and be transferred in culture dish add PBS be heated to paraffin dissolve, be that auxiliary miniature suction pipe can take out required cell to carry out next step research with microscope again, as shown in Figure 6.
Those skilled in the art will readily understand; the foregoing is only preferred embodiment of the present invention; not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. an openable micro-fluidic chip, it is characterized in that, comprise the first substrate and the second substrate, described first substrate has depressed part, described second substrate has lug boss, the lug boss of described second substrate is covered in the depressed part surface of described first substrate, and surrounding is combined with described first substrate by polymer.
2. micro-fluidic chip as claimed in claim 1, it is characterized in that, the lug boss of described second substrate and the width of described first substrate junction are 1mm ~ 2mm.
3. micro-fluidic chip as claimed in claim 1, it is characterized in that, the height of the lug boss of described second substrate is greater than 20 μm.
4. micro-fluidic chip as claimed in claim 1, it is characterized in that, the depressed part of described first substrate is rectangle, and described rectangle reduces gradually in the length direction degree of depth.
5. micro-fluidic chip as claimed in claim 3, it is characterized in that, the degree of depth of the depressed part of described first substrate is 1 μm ~ 200 μm.
6. micro-fluidic chip as claimed in claim 3, it is characterized in that, the length of the depressed part of described first substrate is 50mm ~ 70mm, and width is 10mm ~ 22mm.
7. micro-fluidic chip as claimed in claim 3, is characterized in that, described micro-fluidic chip is provided with entrance and exit, and it is connected with the bosom of the depressed part of described first substrate and the most shallow place respectively.
8. micro-fluidic chip as claimed in claim 1, it is characterized in that, described first substrate and the second substrate are glass.
9. micro-fluidic chip as claimed in claim 1, it is characterized in that, described polymer is dimethyl silicone polymer or epoxy resin.
10., as the preparation method of micro-fluidic chip as described in any one in claim 1-9, it is characterized in that, comprise the following steps:
(1) depressed part is obtained in the first substrate surface etching, lug boss is obtained in the second substrate surface etching, make the lug boss of described second substrate identical with the shape of the depressed part of described first substrate, the profile 1mm ~ 2mm larger than the profile of the depressed part of described first substrate of the lug boss of described second substrate;
(2) lug boss of described second substrate aimed at the depressed part of the first substrate fit, make the lug boss of described second substrate cover the depressed part of described first substrate completely, and with the fitting of described first substrate;
(3) surrounding to the lug boss of described second substrate fills prefabricated polymer, the lug boss of described second substrate is combined with described first substrate, namely obtains described micro-fluidic chip after described polymer cure.
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| CN105363505A (en) * | 2015-12-11 | 2016-03-02 | 武汉纺织大学 | Three-dimensional structural cell capture and release chip and preparation method thereof |
| CN107597219A (en) * | 2017-09-13 | 2018-01-19 | 北京理工大学 | A kind of micro-fluidic chip sample introduction and its bonding techniques |
| CN108671971A (en) * | 2018-05-11 | 2018-10-19 | 北京科技大学 | A kind of circulating tumor cell and the micro fluidic device and method of the separation of cluster feminine gender |
| CN109701298A (en) * | 2018-12-28 | 2019-05-03 | 莆田学院 | Super affine super close-super thin gas copper foil of wetability gradual change and preparation method thereof and Positioning collection device |
| CN110108623A (en) * | 2019-04-30 | 2019-08-09 | 武汉纺织大学 | A kind of greasy dirt grain testing apparatus and method based on micro-fluidic chip |
| CN110643502A (en) * | 2019-10-27 | 2020-01-03 | 苏州济研生物医药科技有限公司 | Single-cell microfluidic detection chip and preparation method and application method thereof |
| EP3612307A1 (en) * | 2017-04-21 | 2020-02-26 | Abaxis, Inc. | Systems, devices and methods for microfluidic analysis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP3612307A1 (en) * | 2017-04-21 | 2020-02-26 | Abaxis, Inc. | Systems, devices and methods for microfluidic analysis |
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| CN109701298A (en) * | 2018-12-28 | 2019-05-03 | 莆田学院 | Super affine super close-super thin gas copper foil of wetability gradual change and preparation method thereof and Positioning collection device |
| CN109701298B (en) * | 2018-12-28 | 2020-12-22 | 莆田学院 | Super-affinity super-hydrophilic-super-hydrophobic gas copper foil with gradually-changed wettability, preparation method thereof and positioning and collecting device |
| CN110108623A (en) * | 2019-04-30 | 2019-08-09 | 武汉纺织大学 | A kind of greasy dirt grain testing apparatus and method based on micro-fluidic chip |
| CN110643502A (en) * | 2019-10-27 | 2020-01-03 | 苏州济研生物医药科技有限公司 | Single-cell microfluidic detection chip and preparation method and application method thereof |
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