CN105536895B - A kind of openable micro-fluidic chip and preparation method thereof - Google Patents
A kind of openable micro-fluidic chip and preparation method thereof Download PDFInfo
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- CN105536895B CN105536895B CN201510915541.1A CN201510915541A CN105536895B CN 105536895 B CN105536895 B CN 105536895B CN 201510915541 A CN201510915541 A CN 201510915541A CN 105536895 B CN105536895 B CN 105536895B
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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
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Abstract
The invention discloses a kind of openable micro-fluidic chip, including the first substrate and the second substrate, first substrate has depressed part, second substrate has lug boss, the lug boss of second substrate is covered in the depressed part surface of first substrate, and surrounding is combined by polymer with first substrate, the polymer is used for the reversible substrate of connection first and the second substrate, so as to form openable micro-fluidic structure.The invention also discloses the preparation method of the micro-fluidic chip.By the invention it is possible to carry out the capture of cell or particulate using micro-fluidic chip, and the cell or particulate for capturing and obtaining are collected, to be further analyzed.
Description
Technical field
The invention belongs to micro-fluidic chip field, more particularly, to a kind of openable micro-fluidic chip and its preparation
Method.
Background technology
Micro-fluidic chip is the core of micro-total analysis system, and the invention of micro-fluidic chip promotes to be applied to chemistry, food, ring
The analytical instrument of border, medical science and life science develops to miniaturization, automation, integrated and portability direction, the advantage is that
Various chemical reactions are detected by micro-fluidic chip, reagent consumption is significantly reduced, and substantially increases analysis efficiency, reduction takes
With.Designing and producing the micro-fluidic chip constituted with microfluidic channel network can be applied to different analysis fields, including chemistry point
The multiple fields such as analysis, food hygiene, environmental monitoring, medical science chemical examination, life science, Forensic science and national defence.
At present in micro-fluidic chip, cell and microparticle are manipulated using photoelectric technology or mechanically actuated more.Light
The instrument that method is related to is complicated, and manipulation is single, limits more.Electrical method generally requires the integrated fine electrode on chip,
Complicated power supply is needed, while voltage on cell has detrimental effect.And mechanically actuated then lack operation flexibility and
Invertibity；For example, conventional such as the micro- dam of cell mechanical positioning element and micropore are fixed due to its adjusting height, cell is once be captured
Then be difficult to it is fully erased, while to capture discharge after next step research there is also certain difficulty.Meanwhile, existing skill
Micro-fluidic chip in art is constituted after being generally bonded with two panels substrate, and substrate is combined very closely, even if capturing cell and micro-
Grain is also difficult to open, to obtain required cell and particulate, or open reluctantly and it cannot be guaranteed that accurate capture, it is difficult to pinpoint
Or orientation obtains the cell or particulate needed for micro-fluidic chip.
The content of the invention
For the disadvantages described above or Improvement requirement of prior art, the invention provides a kind of openable micro-fluidic chip,
Its object is to pass through the substrate of connection first and the second substrate of reversible polymer, so as to form openable micro-fluidic knot
Structure, so as to orient cell or particulate needed for obtaining.
To achieve the above object, according to one aspect of the present invention, there is provided a kind of openable micro-fluidic chip, including
First substrate and the second substrate, first substrate have depressed part, and second substrate has lug boss, second base
The lug boss of piece is covered in the depressed part surface of first substrate, and surrounding is combined by polymer with first substrate,
The polymer is used for the reversible substrate of connection first and the second substrate, so as to form openable micro-fluidic structure.
Preferably, the width of the lug boss of second substrate and the lug boss junction of first substrate for 1mm~
2mm。
Preferably, the height of the lug boss of second substrate is more than 20 μm.
Preferably, the depressed part of first substrate be rectangle, and the depressed part rectangle length direction depth by
Decrescence small, the micro-fluidic chip is used for the capture of cell or particulate.
As it is further preferred that the depth of the depressed part of first substrate is 1 μm~200 μm.
As it is further preferred that first substrate depressed part length be 50mm~70mm, width be 10mm~
22mm。
As it is further preferred that be provided with entrance and exit on the micro-fluidic chip, its respectively with first base
The bosom of the depressed part of piece and most shallow place are connected.
As it is further preferred that the entrance and exit is arranged on the first substrate.
As it is further preferred that the surface of the lug boss of second substrate is coated with 1 μm~5 μm of gel layer.
As it is further preferred that the gel layer is agarose, shitosan or polyvinyl alcohol.
Preferably, first substrate and the second substrate are glass.
Preferably, the polymer is dimethyl silicone polymer or epoxy resin.
It is another aspect of this invention to provide that there is provided a kind of preparation method of above-mentioned micro-fluidic chip, comprising the following steps：
(1) depressed part is obtained in the first substrate surface etching, obtains lug boss in the second substrate surface etching so that described
The lug boss of the second substrate is identical with the shape of the depressed part of first substrate, the profile ratio of the lug boss of second substrate
Big 1mm~the 2mm of profile of the depressed part of first substrate；
(2) lug boss of second substrate is directed at laminating with the depressed part of the first substrate so that second substrate
Lug boss be completely covered the depressed part of first substrate, and fitted with first substrate；
(3) fill prefabricated polymer to the surrounding of the lug boss of second substrate so that second substrate it is convex
The portion of rising is combined with first substrate, and the micro-fluidic chip is obtained after the polymer solidification.
Preferably, between the step (1) and the step (2), also include, in the lug boss table of second substrate
Face covers 1 μm~5 μm of gel layer.
In general, by the contemplated above technical scheme of the present invention compared with prior art, due to by polymer
The reversible substrate of connection first and the second substrate, can obtain following beneficial effect：
1st, it is bulge-structure by the second substrate designs, the surface of the first substrate depressed part is covered in, while using polymer
The circumference of the second substrate lug boss is filled in so that the combination that the first substrate and the second substrate can be reversible, so that convenient open micro-
Fluidic chip is obtaining cell therein or particulate；
2nd, depressed part is preferably set to the uneven rectangle of depth, so that by different size of cell or particle capture in recessed
The different parts in the portion of falling into；
3rd, preferably using glass as the first substrate and the material of the second substrate, because glass is transparent material, cell and micro-
The acquisition procedure of grain is more controllable；
4th, the preparation method of the micro-fluidic chip is simple, efficiency high, is easy to orientation to obtain thin needed for micro-fluidic chip
Born of the same parents and particulate.
Brief description of the drawings
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 prepared using embodiment 1 captures cell schematic diagram；
Fig. 5 is paraffin encapsulation capture cell schematic diagram in micro-fluidic chip prepared by embodiment 1；
Fig. 6 is to shift cell schematic diagram after micro-fluidic chip prepared by embodiment 1 is opened；
In all of the figs, identical reference be used for represent identical element or structure, wherein：The substrates of 1- first,
The substrates of 2- second, 3- chip entrances, the outlet of 4- chips, 5- the first substrate depressed parts, 6- the second substrate lug bosses, 7-PDMS, 8-
Raceway groove bosom, the most shallow place of 9- raceway grooves, 11- marks, 12- cells.
Specific embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, it is right below in conjunction with drawings and Examples
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.As long as additionally, technical characteristic involved in invention described below each implementation method
Not constituting conflict each other can just be mutually combined.
The invention provides a kind of openable micro-fluidic chip, including the first substrate and the second substrate, described first
Substrate has depressed part, and the lug boss that second substrate has lug boss, second substrate is covered in first substrate
Depressed part surface, and surrounding combined by polymer with first substrate, and the width of its junction is 1mm~2mm；It is described
Polymer is dimethyl silicone polymer or epoxy resin, for the reversible substrate of connection first and the second substrate, so as to be formed
Openable micro-fluidic structure.First substrate and the second substrate are preferably glass so that the acquisition procedure of cell and particulate
It is more controllable.The height of the lug boss of the second substrate is more than 20 μm so that the binding site of the second substrate and the first substrate has one
Fixed space, to separate the first substrate and the second substrate using instrument when opening.
Micro-fluidic chip for capturing cell and particulate may be by the design.Cell and particulate are carried out in chip
After capture experiment, opening chip, so as to obtain required cell or particulate.
For example, the depressed part of the first substrate can be designed as into length for 50mm~70mm, width is 10mm~22mm's
Rectangular configuration, the depth of the rectangular configuration is 1 μm~200 μm, and its depth is gradually reduced in length direction so that different-diameter
Cell and particulate can be fixed on the diverse location of rectangle, entrance and exit is provided with micro-fluidic chip, its respectively with it is recessed
The bosom and most shallow place in the portion of falling into are connected.The cell or the size of particulate that its specific depth can be captured as needed are carried out
Design, for example, it is desired to when capturing the circulating tumor cell for being mixed with normal cell, the most shallow place of depth can be designed as into 5 μm, bosom
60 μm are designed as, so as in the diverse location in rectangle length direction, capture obtains 15 μm~30 μm and the cell for differing in size.Can
To cover the gel layer of 1 μm~5 μm of the thickness that the materials such as such as agarose, shitosan, polyvinyl alcohol are formed on lug boss surface, make
During chip use, cell or particulate preferably adhere to one side of lug boss, are easy to further microfluidic control chip to open
When, carry out transfer separation.
After make use of chip and carrying out capture experiment, cell or particulate are typically attached to the side of the second substrate lug boss,
Now the specific open method of the micro-fluidic chip is as follows：
(1) alcohol is passed through by gas (bubble) emptying in micro-fluidic chip from porch；
(2) from porch phosphate buffer solution PBS (formulas：Weigh NaCl 8g, KCl 0.2g, Na2HPO4·12H2O
3.63g,KH2PO4 0.24g, are dissolved in 900ml distilled waters, adjust pH value to 7.4 with hydrochloric acid, add water and are settled to 1L, and normal temperature is preserved
It is standby) alcohol in micro-fluidic chip is emptied；
(3) paraffin is passed through from porch, paraffin is full of micro-fluidic chip；
(4) after after paraffin solidification, the polymeric layer of the second substrate lug boss surrounding is removed with instruments such as blades, separates first
Substrate and the second substrate lug boss peripheral region, you can chip is opened, the cell or micro- being encapsulated in paraffin is obtained
Grain, then the paraffin of cell or particulate containing needed for is separated, heating causes paraffin dissolving, you can cell or particulate needed for taking out
To carry out the research of next step.
The preparation method of the micro-fluidic chip, comprises the following steps：
(1) depressed part is obtained in the first substrate surface etching, obtains lug boss in the second substrate surface etching so that described
The lug boss of the second substrate is identical with the shape of the depressed part of first substrate, the profile ratio of the lug boss of second substrate
Big 1mm~the 2mm of profile of the depressed part of first substrate；
(2) lug boss of second substrate is directed at laminating with the depressed part of the first substrate so that second substrate
Lug boss be completely covered the depressed part of first substrate, and fitted with first substrate；
(3) prefabricated polymer (usually polymer monomer and friendship is filled to the surrounding of the lug boss of second substrate
Join the mixture of agent) so that the lug boss of second substrate is combined with first substrate, is obtained final product after the polymer solidification
To the micro-fluidic chip.
Embodiment one kind can open cavity heights controllable precise cell one side transfer micro flow chip
Step 1 prepares the first substrate
From 75mm long, raceway groove 5 is designed as 60mm long by the slide of 25mm wide as the material of the first substrate 1, wide
15mm, its depth is gradually reduced from left to right, and left side bosom is 60 μm, and the most shallow place in right side is 5 μm, with the side of wet etching
Method is performed etching with the speed of 1mm/min and obtains corresponding pattern.And obtained with the central finishing impression at most shallow place respectively in bosom
Entrance 3 and outlet 4, side view and the top view difference of the first substrate for obtaining are as illustrated in figs. 1A and ib.
Step 2. prepares the second substrate
Second substrate 2 is designed as 61mm × 16mm, the boss 6 that about 80 μm of height, with the side similar with the first substrate
Method prepares the second substrate, and its side view and top view difference are as shown in Figure 2 a and 2 b.
Step 3. configures the agarose solution of 2% mass fraction
In the presence of magnetic stirring apparatus, be well mixed for solution by 60 degree of temperature, then takes 500 with the pipettor of 1000 μ l
The solution of μ l drips and is put into sol evenning machine when agarose also uncolled agglutination on boss, is taken advantage of, and uses 800rpm 20s, 1400rpm
The parameter of 40s gets rid of even, finally can about 2 μm of agarose film with 50 DEG C of drying in thermal station.
Step 4. chip is fitted
The first substrate that will be made is aligned with membrane second substrate, the ditch for making boss be completely covered on the first substrate
Road, stacks and make together together with the first substrate is brought into close contact with the second substrate.
Step 5. polymer solidifies
From prefabricated PDMS, (quality of polymer monomer and crosslinking agent is 10:1), treat that PDMS is embedded into around boss
Afterwards, chip completes after being heated to 85 DEG C and making PDMS crosslinking curings in 10 minutes, the chip side view and top view for obtaining point
Not 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.
The test that controllable cell one side shifts micro flow chip can be carried out with the micro-fluidic chip, its specific experiment process is such as
Under：
(1) alcohol is passed through by gas (bubble) emptying in micro-fluidic chip with the speed of 100 μm/min in chip；
(2) phosphate buffer solution PBS (formulas are passed through with the speed of 100 μm/min in chip：Weigh NaCl 8g, KCl
0.2g, Na2HPO4·12H2O 3.63g,KH2PO4 0.24g, are dissolved in 900ml distilled waters, and pH value is adjusted to 7.4 with hydrochloric acid, plus
Water is settled to 1L, and normal temperature is saved backup) alcohol in raceway groove is emptied；
(3) it is passed through concentration as 10 to the speed in chip with 100 μm/min3/ ml and diameter is in 10 μm~20 μm and size
The target cell not waited, is kept for 2 minutes；
(4) phosphate buffer solution PBS is passed through with the speed of 100 μm/min in chip so that cell is according to the big of itself
It is small to stay at the micro-fluidic chip different depth；
(5) examine under a microscope the position of the stop of cell 11 and do mark 12, as shown in Figure 4；
(6) paraffin is passed through with the speed of 100 μm/min in chip；
(7) treat that paraffin solidifies, then the several positions marked with micro- sem observation, at this moment it can be seen that the position of cellular retention
Put and quantity does not change, as shown in Figure 5；
(8) at this moment, then by chip put into 47 degrees Celsius of water and soak, soften the gel rubber material of bottom, ten minutes
The PDMS around boss is removed with specific purpose tool again afterwards, chip is opened and is obtained with being bonded at the paraffin section of boss face, then arrive
Several positions that basis of microscopic observation is marked, at this moment it can be seen that the position of cellular retention and quantity still do not change
Become；
(9) cell or particulate being encapsulated in paraffin layer can be obtained after paraffin is opened, required cell place is further taken out
Partial paraffin, and be transferred in culture dish and add PBS to be heated to paraffin dissolving, then with miniature suction pipe supplemented by microscope
Required cell is taken out to carry out the research of next step, as shown in Figure 6.
As it will be easily appreciated by one skilled in the art that the foregoing is only presently preferred embodiments of the present invention, it is not used to
The limitation present invention, all any modification, equivalent and improvement made within the spirit and principles in the present invention etc., all should include
Within protection scope of the present invention.
Claims (10)
1. a kind of openable micro-fluidic chip, it is characterised in that including the first substrate and the second substrate, first substrate
With depressed part, the lug boss that second substrate has lug boss, second substrate is covered in the recessed of first substrate
Portion surface is fallen into, and surrounding is combined by polymer with first substrate so that the binding site of the second substrate and the first substrate
There is certain space.
2. micro-fluidic chip as claimed in claim 1, it is characterised in that the lug boss of second substrate and first base
The width of piece junction is 1mm~2mm.
3. micro-fluidic chip as claimed in claim 1, it is characterised in that the height of the lug boss of second substrate is more than 20
μm。
4. micro-fluidic chip as claimed in claim 1, it is characterised in that the depressed part of first substrate is rectangle, and institute
Rectangle is stated to be gradually reduced in length direction depth.
5. micro-fluidic chip as claimed in claim 3, it is characterised in that the depth of the depressed part of first substrate is 1 μm
~200 μm.
6. micro-fluidic chip as claimed in claim 3, it is characterised in that the length of the depressed part of first substrate is 50mm
~70mm, width is 10mm~22mm.
7. micro-fluidic chip as claimed in claim 3, it is characterised in that entrance is provided with the micro-fluidic chip and is gone out
Mouthful, it is connected with the bosom and most shallow place of the depressed part of first substrate respectively.
8. micro-fluidic chip as claimed in claim 1, it is characterised in that first substrate and the second substrate are glass.
9. micro-fluidic chip as claimed in claim 1, it is characterised in that the polymer is dimethyl silicone polymer or epoxy
Resin.
10. as described in any one in claim 1-9 micro-fluidic chip preparation method, it is characterised in that including following step
Suddenly：
(1) depressed part is obtained in the first substrate surface etching, obtains lug boss in the second substrate surface etching so that described second
The lug boss of substrate is identical with the shape of the depressed part of first substrate, and the profile of the lug boss of second substrate is than described
Big 1mm~the 2mm of profile of the depressed part of the first substrate；
(2) lug boss of second substrate is directed at laminating with the depressed part of the first substrate so that second substrate it is convex
The portion of rising is completely covered the depressed part of first substrate, and fitted with first substrate；
(3) prefabricated polymer is filled to the surrounding of the lug boss of second substrate so that the lug boss of second substrate
Combined with first substrate, the micro-fluidic chip is obtained after the polymer solidification.
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105363505B (en) * | 2015-12-11 | 2017-05-31 | 武汉纺织大学 | A kind of cell capture of three-dimensional structure and release chip and preparation method thereof |
| CN110730690A (en) * | 2017-04-21 | 2020-01-24 | 爱贝斯股份有限公司 | Systems, devices, and methods for microfluidic analysis |
| 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 |
| 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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