CN105170206B - A kind of micro-fluidic chip of multiple determination - Google Patents
A kind of micro-fluidic chip of multiple determination Download PDFInfo
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Abstract
The invention discloses a kind of micro-fluidic chip of multiple determination, it is related to micro-fluidic chip medical inspection field.Multiple determination micro-fluidic chip includes gas controlled passageway layer, and organic high molecular polymer film layer, microfluidic channel layer, the integrated bonding sealing-in of layers of material forms chip structure, it is characterised in that：Gas controlled passageway layer includes the top half of sample holes, venthole and two pneumatic micro valves and multiple series connection liquid storage tanks；Organic high molecular polymer film is provided with the hole of sample holes, pneumatic micro valve structure, liquid storage tank and venthole correspondence position size；Microfluidic channel layer sets multiple series connection liquid storage tank the latter half and multiple reaction tanks, connect each liquid storage tank between fluid passage, connect the fluid passage of liquid storage tank and reaction tank.Simple structure of the present invention, it is easy to use, make liquid storage tank and reaction tank independence using pneumatic micro valve, it is to avoid liquid phase mutually pollutes between each reaction tank, improves the precision of sample size, it is ensured that the reliability of testing result.
Description
Technical field
The present invention relates to microfluidic chip analysis field, more particularly to a kind of micro-fluidic chip of multiple determination.
Background technology
Micro-fluidic chip refers to a kind of biology built on one piece several square centimeters of chip also known as chip lab
Or chemical laboratory.It is the basic operation list such as reaction involved in biological and chemical field, separation, culture, sorting, detection
The component of micro-/ nano magnitude is made respectively by unit, is integrated on one piece of chip of very little, network is formed by microchannel, with controllable flow
Body runs through whole system, is used to realize the various functions of standard biologic or chemistry.Carried out for the purpose of biological sample analysis
The reaction such as enzyme reaction, immune response, PCR reactions, ELISA can all be realized on micro-fluidic chip.Due to micro-fluidic core
The characteristics of chip technology has that sample size is small, integrated level is high, easily realizes Automated condtrol and high throughput analysis, on micro-fluidic chip
Carry out sample pre-treatments for biochemical reaction operates more conventional analysis more convenient, quick, with low cost.And, high specific
Biochemical reaction be combined with the powerful separation of micro-fluidic chip, detectability and would indicate that significantly more advantage.
When carrying out multi objective biochemistry detection using micro-fluidic chip, due to the diversity of Testing index, need to be set on chip
Put multiple reaction tanks occurred for detection reaction to detect same sample, how rapidly and efficiently sample to be carried out to refer to more
Ensureing the sample size for independently carrying out and participating in each detection reaction of each detection reaction while mark detection unanimously turns into micro-fluidic
Key link in chip design.
Patent of invention CN102671729 discloses a kind of micro-fluidic chip for multi objective biochemistry detection, the chip bag
Include microfluidic channel, several reaction tanks and pneumatic micro valve, the reagent needed for biochemical reaction is fixed with reaction tank, several are anti-
Ying Chi is connected by microfluidic channel serial or parallel.Although the chip can be realized each anti-by controlling the switch of pneumatic micro valve
Connection and isolation between Ying Chi, but the cross pollution of reaction reagent, sample are easily caused in the flow process of sample feeding
Needing to flow through multiple connected reaction tanks after product injection could be full of each reaction tank, and flow rate of liquid is each variant in passage, leads
There is larger difference in the sample size that each detection reaction is participated between causing the different reaction tank in position, be unfavorable for the accurate of each detection reaction
Control and the accurate analysis of testing result.
Based on background above, the support multi objective detection simultaneously of a kind of simple structure, simple operation is invented, and between each index
In the absence of cross pollution, each reaction tank sample size identical micro-fluidic chip is very necessary.
The content of the invention
It is an object of the invention to provide a kind of micro-fluidic chip of multiple determination, while ensureing to participate in each detection reaction
Sample size is identical and the reaction of each Indexs measure between independently there is no cross contamination, realize it is easy, efficiently, low cost it is many
Indexs measure.
To achieve the above object, the present invention is adopted the following technical scheme that：
A kind of micro-fluidic chip of multiple determination, including gas controlled passageway layer（100）, organic high molecular polymer is thin
Film（200）, microfluidic channel layer（300）, the integrated bonding sealing-in formation chip structure of layers of material, it is characterised in that：Gas control
Channel layer processed includes sample holes（1）, venthole（6）With two pneumatic micro valves（2,3）With multiple series connection liquid storage tank（4）The first half
Point；Organic high molecular polymer film is provided with sample holes（1）, pneumatic micro valve structure（2,3）, liquid storage tank（4）And venthole（6）
The hole of correspondence position size；Microfluidic channel layer sets multiple series connection liquid storage tanks（4）The latter half and multiple reaction tanks（9）, even
Lead to each liquid storage tank（4）Between fluid passage（7）, connection liquid storage tank（4）And reaction tank（9）Fluid passage（8）.
Two described micro-valves（2,3）Including valve seat（10,12）With micro-valve gas channels（11,13）, by controlling air pressure,
Control micro-valve open and close, and then liquid flow direction in microfluidic channel layer is controlled, after outside air enters micro-valve, extrude intermediate layer
Film is allowed to raised and blocks lower channel, reaches control effect；Wherein micro-valve（2）Tool valve seat（10）With multiple branches gas channels
（11）, passage is intersecting to be located at each liquid storage tank（4）Interface channel top, controls each liquid storage tank（4）Between fluid passage（7）Switch；It is micro-
Valve（3）Tool valve seat（12）With gas channels normal thereto（3）, it is intersecting to be arranged at liquid storage tank（4）And reaction tank（9）Interface channel
（8）On, control interface channel（8）Switch.
The pneumatic micro valve（2,3）Valve seat（10）With（12）Can be connected with the outer source device such as gas bomb or syringe
It is logical, and then to the micro-valve gas channels（11）With（13）Interior offer positive and negative pressure.
Described liquid storage tank（4）For size is identical, parallel series connection, fluid passage（7）Topmost accessed by it and picked out, most
End Connection Pool（5）, store excessive sample.
Described micro-fluidic chip material may be selected glass, dimethyl silicone polymer（PDMS）, polymethyl methacrylate
（PMMA）, polyethylene terephthalate（PET）Or makrolon（PC）Middle any one.
Described sample reservoir（4）The material of upper half side elastic film can be dimethyl silicone polymer（PDMS）, silicon
Rubber, memorial alloy or polytetrafluoroethylene (PTFE).
The reaction tank（9）It is interior in advance be fixed with multiple determination needed for reagent, such as enzyme and substrate, antigen or antibody
And PCR primer etc., can be embedded by physical absorption, chemical crosslinking, sol-gel, microballon be fixed or the mode such as film is fixed
It is fixed.
A kind of method of work of the micro-fluidic chip of multiple determination, comprises the following steps：
Step one：During sample-adding, micro-valve（2）Open, micro-valve（3）Close.By valve seat（12）To micro-valve gas channels（13）
Gas-pressurized is inside passed through, liquid storage tank is closed to reaction tank direction fluid passage.Sample is from sample holes（1）Injection, sample flows through liquid
Body passage（7）Flow into each sample liquid storage tank（4）In, unnecessary sample flows into Sample Buffer pond（5）In.
Step 2：When each sample liquid storage tank（4）When being each filled with, micro-valve（2）Close, micro-valve（3）Open, by valve seat（10）
To micro-valve gas channels（11）Inside it is passed through gas-pressurized, each liquid storage tank of closing control（4）Between fluid passage（7）Interior liquid flowing
Micro-valve, realizes each sample liquid storage tank（4）Between isolation；Open the fluid passage of control each sample liquid storage tank and corresponding reaction tank
Micro-valve, press each sample liquid storage tank（4）The elastic film of top, makes sample from each sample liquid storage tank（4）Middle inflow is corresponding
Reaction tank（9）In, sample flows into reaction tank（9）Afterwards be fixed on reaction tank in advance（9）In multiple determination carried out with reagent
Reaction.
Beneficial effect
The invention provides a kind of micro-fluidic chip of multiple determination, based on microfluidic chip technology platform, collect sample
Sample introduction, multiple determination need to only combine sampling device and inject sample in one, during detection, using additional source of the gas control micro-valve
Open and close, manual or other modes pressing cavity wall is to be capable of achieving detection, and simple structure is easy to use, with low cost；Utilize
The sample reservoir of the formed objects of multiple serial connection makes the sample size that each detection of participation is reacted identical, and sample is stored and reaction
Different regions are located at, each reaction tank is formed independent conversion zone using integrated pneumatic micro valve, it is to avoid sample feeding
And by the liquid reagent contamination that causes of flowing and interfered in course of reaction, improve the essence for participating in detection response sample amount
Degree, it is ensured that the reliability of testing result.
Brief description of the drawings
Fig. 1 multiple determination chip overall schematics.
Top tank air control passage Rotating fields schematic diagram in Fig. 2 multiple determination chips.
Intermediate layer PDMS film structural representation in Fig. 3 multiple determination chips.
Lower floor's microfluidic channel Rotating fields schematic diagram in Fig. 4 multiple determination chips.
Overall structure composition schematic diagram in Fig. 5 multiple determination chips.
Micro-valve in Fig. 6 multiple determination chips opens and closes schematic diagram.
1 is sample holes, and 2 is liquid storage tank pneumatic micro valve, and 3 is reaction tank pneumatic micro valve, and 4 is sample reservoir, and 5 is sample
Buffer Pool, 6 is venthole, and 7 is liquid storage tank fluid passage, and 8 is reaction tank fluid passage, and 9 is reaction tank, and 10 is liquid storage tank micro-valve
Valve seat, 11 liquid storage tank micro-valve gas channels, 12 is reaction tank micro-valve valve seat, and 13 is reaction tank micro-valve gas channels, and 100 is gas
Control passage layer, 200 is organic high molecular polymer film, and 300 is microfluidic channel layer.
Specific embodiment
The present invention will be further described for following example, and the example is only one of implementation method of apparatus of the present invention, and
Any limitation is not made to the present invention.
Embodiment one：Microfluidic chip structure
As shown in Fig. 1,2,3,4,5：A kind of micro-fluidic chip for the detection of many biochemical indicators, by the core for being integrated with micro-valve
Sheet material bonding sealing-in is formed, including gas controlled passageway layer 100, organic high molecular polymer film 200, microfluidic channel layer
300, wherein：
Gas controlled passageway layer 100 is provided with sample feeding hole 1,6, two pneumatic micro valves 2,3 of venthole and multiple series connection
The top half of sample reservoir 4, the side of the top half of sample reservoir 4 is for elastic film and in air；
Microfluidic channel layer 300 includes the fluid passage 7 of connection sample holes and each sample liquid storage tank, connects each sample liquid storage
The latter half of pond and the fluid passage 8, sample reservoir of corresponding reaction tank, the sample connected with sample injection channel end
Buffer Pool 5 and reaction tank 9, are fixed with the reaction reagent of multiple determination in reaction tank；
Organic high molecular polymer film 200 is sandwiched between gas controlled passageway layer 100 and microfluidic channel layer 300, thin
The position of correspondence sample holes 1, valve seat 10 and 12, venthole 6 and sample reservoir 4 is provided with correspondingly sized hole on film；
In the above-mentioned micro-fluidic chip for multiple determination, the material of described micro-fluidic chip is PC, PDMA,
One kind in PMMA, PET, the present embodiment the selection of material is PC, and the thickness of control passage layer is 3 ~ 5mm, the thickness of fluid passage layer
It is 120 ~ 180um to spend（Can change different-thickness size according to actual conditions）；
In the above-mentioned micro-fluidic chip for multiple determination, the bullet of the top half side wall of the sample reservoir 4
Property film material be silicon rubber, PDMS, the one kind in memorial alloy or polytetrafluoroethylene (PTFE), the present embodiment selects silicon rubber, bullet
Property film thickness be 60 ~ 100um；
In the above-mentioned micro-fluidic chip for multiple determination, valve seat 2 and 11 and the gas of the pneumatic micro valve 12 are noted
Emitter is connected, and then can make film 14 that elastic deformation to occur to gas-pressurized is passed through in the micro-valve gas channels 3 and 10,
The thickness of film is 60 ~ 100um；
Be fixed with advance respectively in the above-mentioned micro-fluidic chip for multiple determination, in 4 reaction tanks 9 for
Enzyme and substrate needed for 4 kinds of biochemical indicator detections；
The cross section of sample holes is circle, a diameter of 0.75mm；The rank street face of venthole is circle, a diameter of 0.3mm；Sample
The cross section of product liquid storage tank is circle, and a diameter of 7.5mm is highly 0.2mm；The cross section of reaction tank is ellipse, a diameter of
15mm × 8mm, is highly 0.2mm；Sample Buffer pond is rectangle, and 10mm long, 6mm wide are highly 0.2mm；Fluid passage is high
100um, 300um wide, cross section are semicircle；Gas controlled passageway 100um high, 300um wide, cross section is semicircle, above-mentioned
Data are only limitted to the present embodiment, and the invention is not limited in data in the present embodiment, data are according to actual feelings in other embodiments
Condition can suitably adjust change.
Embodiment two：Micro-fluidic chip workflow
Step one：During sample-adding, micro-valve 2 is opened, and micro-valve 3 is closed, by valve seat 12 to being passed through in micro-valve gas channels 13 plus
Calm the anger body, close liquid storage tank to reaction tank direction fluid passage, sample injects from sample holes 1, and sample flows through fluid passage 7 and flows into
In each sample liquid storage tank 4, unnecessary sample is flowed into Sample Buffer pond 5；
Step 2：When each sample liquid storage tank 4 is each filled with, micro-valve 2 is closed, and micro-valve 3 is opened, by valve seat 10 to micro-valve gas
Gas-pressurized is passed through in paths 11, the micro-valve of liquid flowing, realizes various kinds in closing control 4 fluid passages 7 of each liquid storage tank
Isolation between product liquid storage tank 4；The micro-valve of control each sample liquid storage tank and the fluid passage of corresponding reaction tank is opened, various kinds is pressed
The elastic film of the top of product liquid storage tank 4, in sample is flowed into corresponding reaction tank 9 from each sample liquid storage tank 4, sample flows into anti-
Reacted with reagent with the multiple determination being fixed in advance in reaction tank 9 after answering pond 9.
Embodiment three：Facture of microchip method
（One）Prepare the mould with micro-structural on micro-fluidic chip
Microchannel and the micro-structural of micro-fluidic chip are designed with computer aided design software, according to design drawing chromium plate point
Not Zhi Zuo gas controlled passageway layer and microfluidic channel layer mask.
Photoetching is carried out from silicon chip and etching makes mould, first, Wafer Cleaning is clean, the silicon chip nitrogen for cleaning up
It is put on sol evenning machine after air-blowing is dry, takes appropriate photoresist and be applied on silicon chip, start sol evenning machine spin coating photoresist, mask plate is put
It is placed in litho machine after on the silicon chip for scribble photoresist and is exposed, dry etching is utilized after development dries technique with after
Obtain the mould of the micro-structural with top tank air control passage layer and lower floor's microfluidic channel layer.
（Two）Pressure sintering makes chip
Will be a kind of in PDMS, PMMA, PET, PC sheet material（The present embodiment selects PC）Small pieces are cut into by die size, are cleaned, dried
It is placed on mould after dry, at 180 DEG C on hot press, plus 1000lb pressure 4.5min, the demoulding, be obtained has microfluidic channel respectively
With the PC chips of gas controlled passageway.
（Three）By dimethyl silicone polymer（PDMS）, it is silicon rubber, a kind of in memorial alloy or polytetrafluoroethylene (PTFE)（The present embodiment
Use PDMS）Performed polymer and curing agent are by volume 10：1 ratio mixing, by mixed liquor pour with chip formed objects
On several, glass, heat up after being crosslinked 2 hours, it is the PDMS film of 80um to form the good thickness of elasticity.
（Four）With ultrasonic drilling machine in step（Two）The top tank air control passage layer and lower floor's microfluidic channel for obtaining
Relevant position on layer PC chips carries out ultrasonic drilling, sample holes, valve seat and venthole is obtained, in step（Three）Obtained
Ultrasonic drilling on PDMS film, obtains and sample holes, sample reservoir and outlet hole site size identical hole.
（Five）Step（Four）The chips of PC up and down for obtaining are put into plasma cleaner vacuum and bombard 1min, micro- in lower floor rapidly
The reagent of each reaction is loaded in reaction tank on fluid channel layer, rapidly by step（Four）The PDMS for obtaining is placed in top tank air
Between control passage layer and lower floor's microfluidic channel layer, by step（Three）The PDMS film for obtaining is cut to be stored up with diameter greater than sample
Liquid pool size, the correspondence position being placed on top tank air control passage layer, carries out irreversible sealing-in and obtains being integrated with the micro- of micro-valve
Fluidic chip.
Claims (7)
1. a kind of micro-fluidic chip of multiple determination, including gas controlled passageway layer（100）, organic high molecular polymer film
（200）, microfluidic channel layer（300）, the integrated bonding sealing-in formation chip structure of layers of material, it is characterised in that：Gas is controlled
Channel layer（100）Including sample holes（1）, venthole（6）With liquid storage tank pneumatic micro valve（2）, reaction tank pneumatic micro valve（3）And it is many
Individual series connection sample reservoir（4）Top half；Organic high molecular polymer film（200）It is provided with sample holes（1）, liquid storage tank
Pneumatic micro valve（2）, reaction tank pneumatic micro valve（3）, sample reservoir（4）And venthole（6）The hole of correspondence position size；Microfluid
Channel layer（300）Multiple series connection sample reservoirs are set（4）The latter half and multiple reaction tanks（9）, connection each sample liquid storage tank
（4）Between liquid storage tank fluid passage（7）, connection liquid storage tank（4）And reaction tank（9）Reaction tank fluid passage（8）；Described storage
Liquid pool pneumatic micro valve（2）, reaction tank pneumatic micro valve（3）Include corresponding liquid storage tank micro-valve valve seat respectively（10）, reaction tank micro-valve
Valve seat（12）With liquid storage tank micro-valve gas channels（11）, reaction tank micro-valve gas channels（13）, by controlling air pressure, control micro-valve
Open and close, and then control microfluidic channel layer（300）Middle liquid flow direction, after outside air enters micro-valve, extruding intermediate layer is thin
Film is allowed to raised and blocks lower channel, reaches control effect；Wherein liquid storage tank pneumatic micro valve（2）With liquid storage tank micro-valve valve seat
（10）With multiple branches liquid storage tank micro-valve gas channels（11）, passage is intersecting to be located at each sample liquid storage tank（4）Fluid passage top,
Control each sample liquid storage tank（4）Between liquid storage tank fluid passage（7）Switch；Reaction tank pneumatic micro valve（3）With reaction tank micro-valve
Valve seat（12）With reaction tank micro-valve gas channels normal thereto（13）, it is intersecting to be arranged at sample reservoir（4）And reaction tank（9）
Reaction tank fluid passage（8）On, control reaction tank fluid passage（8）Switch.
2. the micro-fluidic chip of multiple determination according to claim 1, it is characterised in that：The liquid storage tank pneumatic micro valve
（2）, reaction tank pneumatic micro valve（3）Liquid storage tank micro-valve valve seat（10）With reaction tank micro-valve valve seat（12）With gas bomb or injection
Device is connected, and then to the liquid storage tank micro-valve gas channels（11）With reaction tank micro-valve gas channels（13）It is interior to provide positive and negative
Pressure.
3. the micro-fluidic chip of multiple determination according to claim 1, it is characterised in that：Described sample reservoir
（4）Size is identical, parallel series connection, liquid storage tank fluid passage（7）Most gone up along accessing and picking out by it, end connects Sample Buffer pond
（5）, store excess sample.
4. the micro-fluidic chip of multiple determination according to claim 1, it is characterised in that：Described micro-fluidic chip material
Matter selection glass, dimethyl silicone polymer（PDMS）, polymethyl methacrylate（PMMA）, polyethylene terephthalate
（PET）Or makrolon（PC）Middle one kind.
5. the micro-fluidic chip of multiple determination according to claim 1, it is characterised in that：Described sample reservoir
（4）The material of upper half side elastic film is dimethyl silicone polymer, silicon rubber, memorial alloy or polytetrafluoroethylene (PTFE).
6. the micro-fluidic chip of multiple determination according to claim 1, it is characterised in that：The reaction tank（9）It is interior pre-
Reagent needed for being first fixed with multiple determination is selected from the one kind in enzyme and substrate, antigen or antibody and PCR primer, by thing
Reason is adsorbed, is chemically crosslinked, sol-gel is embedded, microballon is fixed or film fixed form is fixed.
7. a kind of method of work of the micro-fluidic chip of multiple determination as claimed in claim 1, it is characterised in that：Including such as
Lower step：
Step one：During sample-adding, liquid storage tank pneumatic micro valve（2）Open, reaction tank pneumatic micro valve（3）Close, by reaction tank micro-valve
Valve seat（12）To reaction tank micro-valve gas channels（13）Gas-pressurized is inside passed through, sample reservoir is closed（4）To reaction tank（9）Side
To reaction tank fluid passage（8）, sample is from sample holes（1）Injection, sample flows through liquid storage tank fluid passage（7）Flow into each sample storage
Liquid pool（4）In, unnecessary sample flows into Sample Buffer pond（5）In；
Step 2：When each sample liquid storage tank（4）When being each filled with, liquid storage tank pneumatic micro valve（2）Close, reaction tank pneumatic micro valve（3）
Open, by liquid storage tank micro-valve valve seat（10）To liquid storage tank micro-valve gas channels（11）Gas-pressurized is inside passed through, closing control is each
Sample reservoir（4）Between liquid storage tank fluid passage（7）The micro-valve of interior liquid flowing, realizes each sample liquid storage tank（4）Between isolation；
Open control each sample liquid storage tank（4）With corresponding reaction tank（9）Reaction tank fluid passage（8）Micro-valve, press each sample
Liquid storage tank（4）The elastic film of top, makes sample from each sample liquid storage tank（4）It is middle to flow into corresponding reaction tank（9）In, sample stream
Enter reaction tank（9）Afterwards be fixed on reaction tank in advance（9）In multiple determination reacted with reagent.
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| CN2831115Y (en) * | 2005-05-19 | 2006-10-25 | 复旦大学 | Multi-channel microflow controlled chip |
| CN102614948A (en) * | 2012-04-05 | 2012-08-01 | 北京金智捷生物科技有限公司 | Micro-fluidic chip and manufacturing method thereof |
| CN102671729A (en) * | 2012-05-07 | 2012-09-19 | 博奥生物有限公司 | Micro-fluidic chip for multi-index biochemical detection |
| CN203842597U (en) * | 2014-05-26 | 2014-09-24 | 博奥生物集团有限公司 | Micro-fluidic chip with deformable membrane capable of preventing reflux |
| CN205127987U (en) * | 2015-09-24 | 2016-04-06 | 基蛋生物科技股份有限公司 | Micro -fluidic chip for multi -index detection |
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| CN109731620A (en) * | 2018-12-29 | 2019-05-10 | 哈尔滨工业大学 | A kind of Pneumatic horizontal micro-fluidic biological biomedical chip with transition cavity |
| CN109731620B (en) * | 2018-12-29 | 2021-01-12 | 哈尔滨工业大学 | Pneumatic horizontal micro-fluidic biomedical chip with transition cavity |
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