CN105289763A - Multi-index detection micro-fluidic chip capable of quantitatively shunting - Google Patents
Multi-index detection micro-fluidic chip capable of quantitatively shunting Download PDFInfo
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Abstract
The invention discloses a multi-index detection micro-fluidic chip capable of quantitatively shunting and relates to the field of medical detection of micro-fluidic chips. The multi-index detection micro-fluidic chip comprises a substrate and a cover piece, wherein the substrate and the cover piece are bonded to form a sheet-shaped chip; a sample inlet is formed in the cover piece and the substrate is provided with a plurality of grades of shunting branch circuits; each shunting branch circuit is communicated with one end of a corresponding parallel reaction cavity; the sizes of the reaction cavities are the same; the other ends of the shunting branch circuits are communicated with a plurality of buffering tanks through liquid buffering channels; the buffering tanks are connected in series; the sizes of the buffering tanks are gradually reduced along a liquid flowing direction; the tail ends of the liquid buffering channels are communicated with an air outlet formed in the cover piece on the upper layer. When samples are added, by gradually reducing the negative-pressure effect of the buffering tanks, a sample amount in each parallel reaction cavity is accurately controlled and errors between the sample amounts participating in a reaction in each index are reduced. The chip has a simple structure and can be flexibly added according to the requirements on the parallel reaction cavities, so that a multi-index detection reaction is realized and the sample amounts of the reaction cavities are the same; the reaction precision is greatly improved and a result is accurate and reliable.
Description
Technical field
The present invention relates to a kind of liquid shunt device, particularly relate to the part flow arrangement of sample size in accurate quantification reaction chamber in a kind of realized multiple determination be located on micro-fluidic chip.
Background technology
In the micro-fluidic chip of microfluid, microchannel size is in micro nano structure, chip material such as glass, silicon chip, macromolecule polymer material etc. mostly are hydrophobic material, the degree of roughness of microchannel surface is also not quite similar, these factors cause micro-size effect, skin effect and wall slippage effect, and these effects all can impact drag size suffered during microfluidic flow.There is certain difference in microchannel surface characteristics such as each interchannel size, degree of roughness in process, makes the rate of flow of fluid of microfluid in microchannel uneven, and then when carrying out multichannel shunting, each interchannel microfluidic flow property of there are differences.
When multiple determination is carried out to same sample, same sample needs to carry out multiple detection reaction, identical in the example reaction amount realizing needing while sample is once distributed to multiple reaction chamber to control between each reaction, ensure that the sample detection amount in each reaction chamber is unanimously conducive to carrying out the control of course of reaction and the analysis of reaction result, reduce the impact of the error in each reaction chamber between sample size on reaction result.
Patent of invention CN102671729 discloses a kind of micro-fluidic chip for multi objective biochemistry detection, this chip comprises microfluidic channel, several reaction tanks and pneumatic micro valve, reaction tank internal fixtion has the reagent needed for biochemical reaction, several reaction tanks are connected in series or parallel join by microfluidic channel, each reaction chamber can be isolated by the control channel in pneumatic micro valve, and the openend of control channel and gas bomb or syringe are connected.Though this chip by utilizing the isolation of micro-valve to realize all being full of equivalent sample in each reaction tank after once injecting enough samples in formed objects reaction tank connected in series, but the sample size of the participation reaction that the interference between each reaction tank that sample causes in lasting flow process causes in diverse location reaction tank produces comparatively big error, also can cause mixing and polluting between each reaction, be unfavorable for the accurate analysis of reaction result.
A kind of multi-channel structure that sample can be branched to multiple detection reaction chamber is disclosed in patent of invention US20150093760, be provided with the injection port being communicated with many fluid passages, each fluid passage end is connected with the identical circular reaction chamber of a size supplying Indexs measure reaction to occur, reaction chamber internal fixtion has Indexs measure reagent, be communicated with from injection port in this invention structure many fluid passages different with the distance of injection port, sample is subject to the impact of dimensional effect that MCA causes and wall slippage effect in flow process, there will be in each reaction chamber to reach and be full of sample simultaneously, final sample size difference of participating in reaction in each reaction chamber when causing detecting, affect the accurate analysis of testing result, ensure all to be full of in each reaction chamber sample and then must control the sample injection time that gross sample amount reaches excessive and enough, be unfavorable for efficiently carrying out and easily causing sample waste and pollution of reaction.
Summary of the invention
For solving problems of the prior art, the invention provides a kind of part flow arrangement that can be used for quantitative sample size in multiple determination, single injected sampling can realize the reaction of multiple Indexs measure and can ensure that the sample detection amount in each reaction chamber reaches consistent, ensure that gross sample participates in each amount of samples of reacting in parallel reaction chamber after shunting identical, be convenient to the generation and the analysis that accurately control reaction.
The technical scheme that the present invention takes is as follows:
A kind of multiple determination micro-fluidic chip of quantitative shunting, comprise underlying basal (4) and upper strata cover plate (2), bonding forms sheet chip, it is characterized in that cover plate (2) is provided with injection port (1), substrate (4) is provided with multi-stage diffluence branch road (5) and reaction chamber, the one end of the reaction chamber (6) that size is identical is communicated with corresponding each diverter branch (5), the other end is communicated with multiple Buffer Pool (8) by fluid cushion passage (7), contact between Buffer Pool (8), the size of Buffer Pool (8) reduces successively with liquid flow direction, fluid cushion passage (7) end is connected with the gas outlet (3) on cover plate (2).
Described parallel reaction chamber (6) has multiple, and be preferably 4-12, reaction chamber fixes multiple determination reagent in (6) in advance, arranges corresponding reaction chamber size and shape according to differential responses.
Described injection port (1) can be set to different shapes and size according to different sample adding devices or application of sample amount.
The sum of series every grade runner branches of described diverter branch (5) is several to be needed to arrange flexibly according to difference, and preferred progression is 2-4 level, and runner branches number is 4-12.
Described fluid cushion passage (7) is in arcuation, and its radian and length adjust according to sample size size in reaction chamber.
Described Buffer Pool (8) reduces successively along liquid flow direction, and Buffer Pool (8) quantity and size be total amount and the change of part flow arrangement space allocation per sample.
The each section of fluid cushion passage (7) that described parallel reaction chamber (6) and Buffer Pool (8) are communicated with all is accessed from the upper edge of parallel reaction chamber (6) and Buffer Pool (8) and is picked out, the width of diverter branch is greater than fluid cushion passage (7) width, produces dimensional effect and then produce negative pressure in sample flow process.
Described substrate (4) and the Material selec-tion dimethyl silicone polymer (PDMS) of cover plate (2), polymethyl methacrylate (PMMA), Merlon (PC), in any one, realize the surface hydrophilicity modification of chip after described substrate (4) and cover plate (2) micro Process arrange corresponding injection port, reaction chamber, buffer channel, Buffer Pool and gas outlet through surface treatment, rear bonding realizes airtight sealing-in.
Beneficial effect
The present invention is compared to the advantage of prior art: when carrying out sample shunting, total sample size of sample is allowed to more than the sample size summation in each parallel reaction chamber, and the scope of gross sample amount when expanding shunting, is easy to operation.When gross sample amount is excessive, precisely controls the sample size in the rear each parallel reaction chamber of shunting, reduce the error participated in isolated each Indexs measure between the sample size of reaction.Sample storage unnecessary after shunting in apparatus of the present invention, in each Buffer Pool, avoids the pollution of sample, conveniently carries out liquid waste processing.This shunting chip apparatus structure is simple, be easy to control, easy to use, diverter branch can add flexibly according to the demand in parallel reaction chamber, single injected sampling can realize the reaction of multiple Indexs measure and can ensure that the sample detection amount in each reaction chamber is identical, improve the accuracy of reaction, be conducive to the accurate analysis of multiple determination result.
Accompanying drawing explanation
Fig. 1 is the upper strata cover plate schematic diagram of the part flow arrangement that can be used for four kinds of Indexs measure.
Fig. 2 is the underlying basal schematic diagram of the part flow arrangement that can be used for four kinds of Indexs measure.
Fig. 3 is the overall structure top view of the part flow arrangement that can be used for four kinds of Indexs measure.
Fig. 4 is the overall structure top view of the part flow arrangement that can be used for six kinds of Indexs measure.
Fig. 5 is the overall structure top view of the part flow arrangement that can be used for eight kinds of Indexs measure.
1 is injection port, and 2 is upper strata cover plate, and 3 is gas outlet, and 4 is underlying basal, and 5 is diverter branch, and 6 is parallel reaction chamber, and 7 is fluid cushion passage, and 8 is Buffer Pool.
Detailed description of the invention
The present invention will be further described for example below, and this example is only one of embodiment of apparatus of the present invention, does not make any restriction to the present invention.
Embodiment one: multi objective microfluidic chip structure
As shown in Figure 1, 2, 3, a kind of part flow arrangement for four kinds of Indexs measure, can branch to sample size in four reaction chambers by accurate quantification, this part flow arrangement is the plates by upper strata cover plate 2 and underlying basal 4 bonding.Upper strata cover plate 2 is provided with injection port 1, underlying basal 4 is provided with the multi-stage diffluence branch road 5 be communicated with sample feeding mouth, each diverter branch 5 is connected with the one end in corresponding parallel reaction chamber 6, the size in parallel reaction chamber 6 is identical, the other end in parallel reaction chamber is communicated with multiple Buffer Pool 8 by arcuation fluid cushion passage 7, connect in string shape between Buffer Pool 8, size reduces successively along with liquid flow direction, and fluid cushion passage 7 end is connected with the gas outlet 3 on the cover plate of upper strata.
In the present embodiment, parallel reaction chamber number is 4,6,8 (shown in Fig. 1,2,3,4,5), in practice according to response situation, between the preferred 4-12 of reaction chamber number, multiple determination reagent is fixed in advance in simultaneous reactions chamber, arrange corresponding reaction chamber size and shape according to differential responses, usual diverter branch adopts T-shaped fluid passage.
In the present embodiment, preferred reaction cavity shape is circular, and each reaction chamber amount of capacity is consistent, diameter between 6-12mm, height 0.5mm.
In the present embodiment, injection port 1 cross section of device is circular, and aperture is 4mm, is set to different shapes and size in practice according to different sample adding devices or application of sample amount.
In the present embodiment, each section of arcuation fluid cushion passage 7 of four parallel reaction chambeies 6 and two Buffer Pool 8 connections all accesses from the upper edge of parallel reaction chamber 6 and Buffer Pool 8 and picks out,
Being used for the width of the diverter branch at different levels 5 carrying out sample shunting in the present embodiment is greater than the fluid cushion passage 7 of arcuation, the width of diverter branch 5 is 300um, the degree of depth is 100um, the width of fluid cushion passage is 200um, the degree of depth is 100um, in other embodiments, diverter branch and buffering channel width, the degree of depth suitably change according to different demand.Fluid cushion passage is arcuation, and its radian and length adjust according to sample size size in reaction chamber.
Buffer Pool 8 in each diverter branch 5 in the present embodiment is in connected in series; be provided with two Buffer Pools; cross section is circle; highly be 0.5mm; the diameter of first forward Buffer Pool is 4mm; the diameter of second Buffer Pool is 2mm (be only limitted to the present embodiment, other embodiments change according to demand voluntarily, still within scope).
The each section of fluid cushion passage that parallel reaction chamber in microfluidic chip structure of the present invention and Buffer Pool are communicated with all accesses from the upper edge of parallel reaction chamber and Buffer Pool and picks out, the width of diverter branch is greater than fluid cushion channel width, produces dimensional effect and then produce negative pressure in sample flow process.
This part flow arrangement selects any one in the macromolecule polymeric materials such as PDMS, PMMA, PC as the material of chip apparatus, micro Process arranges the surface hydrophilicity modification that corresponding injection port 1, reaction chamber 6, buffer channel 7, Buffer Pool 8 and the process of rear surface, gas outlet 3 realize chip, and bonding realizes airtight sealing-in.
Embodiment two: micro-flow control chip preparation method
Step one: by microchannel and the micro-structural of computer aided design software design part flow arrangement chip, is desired to make money or profit in the chip structure road of drafting with high-resolution printers using graphic printing on SU-8 glue as mask, adopt standard photolithography process to make mould, obtain the SU-8 formpiston of tool part flow arrangement upper strata cover plate and underlying basal microchannel and micro-structural.
Step 2: by PDMS performed polymer and curing agent 10:1 mixing in mass ratio, bleed in vacuum drying chamber, be poured over the die surface that step one obtains, take out after 80 DEG C of baking 1h, cooling is slow afterwards peels off PDMS and mould, gets out sample feeding mouth and gas outlet with special drilling tool in relevant position.
Step 3: the upper strata cover plate after cutting into suitable size and underlying basal are put into plasma cleaner vacuum bombardment 1min, rapidly two-layer PDMS is aimed at laminating, carry out irreversible involution, obtain part flow arrangement chip.
Embodiment three: micro-fluidic chip workflow
The sample solution needing to carry out shunting is injected from sample feeding mouth 1, the upper edge of the one end from each parallel reaction chamber 6 after sample solution flows through diverter branch 5 at different levels flows into, when sample is full of reaction chamber, in the sample arcuation fluid cushion passage 7 that edge picks out on the reaction chamber other end Buffer Pool 8 that slowly inflow size reduces gradually; The each section of arcuation fluid cushion passage 7 be communicated with parallel reaction chamber 6 and Buffer Pool 8 all accesses from the upper edge of reaction chamber 6 and Buffer Pool 8 and picks out, and the sample in reaction chamber 6 can not be flowed out when underfill.In sample flow process, the parallel reaction chamber 6 larger from volume flows into the Buffer Pool 8 of small volume through fluid passage, width is less than the Buffer Pool 8 that the fluid cushion passage 7 of the arcuation of diverter branch 5 and size connected in series reduce successively and produces dimensional effect when sample flow, negative pressure is formed in passage, slow down the speed that sample flows into this sample cavity, sample is made to continue in sample introduction branching process, sample is made to branch in other parallel reaction chamber when being full of sample in this sample cavity of guarantee, solve the sample caused by each interchannel sample flow speed difference and shunt uneven problem, sample flow in each diverter branch all follows this rule, shunting terminate after, in each parallel reaction chamber sample size identical.
Claims (8)
1. the multiple determination micro-fluidic chip quantitatively shunted, comprise underlying basal (4) and upper strata cover plate (2), bonding forms sheet chip, it is characterized in that cover plate (2) is provided with injection port (1), substrate (4) is provided with multi-stage diffluence branch road (5) and reaction chamber, the one end of the reaction chamber (6) that size is identical is communicated with corresponding each diverter branch (5), the other end is communicated with Buffer Pool (8) by fluid cushion passage (7), contact between Buffer Pool (8), the size of Buffer Pool (8) reduces successively with liquid flow direction, fluid cushion passage (7) end is connected with the gas outlet (3) on cover plate (2).
2. micro-fluidic chip according to claim 1, it is characterized in that described reaction chamber (6) has multiple, be preferably 4-12, reaction chamber fixes multiple determination reagent in (6) in advance, arranges corresponding reaction chamber size and shape according to differential responses.
3. micro-fluidic chip according to claim 1, is characterized in that described injection port (1) can be set to different shapes and size according to different sample adding devices or application of sample amount.
4. micro-fluidic chip according to claim 1, it is characterized in that the sum of series every grade runner branches of described diverter branch (5) is several and arrange flexibly according to difference needs, preferred progression is 2-4 level, and runner branches number is 4-12.
5. micro-fluidic chip according to claim 1, it is characterized in that described fluid cushion passage (7) is in arcuation, its radian and length adjust according to sample size size in reaction chamber.
6. micro-fluidic chip according to claim 1, is characterized in that described Buffer Pool (8) reduces successively along liquid flow direction, and Buffer Pool (8) quantity and size be total amount and the change of part flow arrangement space allocation per sample.
7. micro-fluidic chip according to claim 1, it is characterized in that each section of fluid cushion passage (7) that described parallel reaction chamber (6) and Buffer Pool (8) are communicated with all is accessed from the upper edge of parallel reaction chamber (6) and Buffer Pool (8) and picked out, the width of diverter branch is greater than fluid cushion passage (7) width, produces dimensional effect and then produce negative pressure in sample flow process.
8. micro-fluidic chip according to claim 1, it is characterized in that the Material selec-tion dimethyl silicone polymer (PDMS) of described substrate (4) and cover plate (2), polymethyl methacrylate (PMMA), Merlon (PC), in any one, realize the surface hydrophilicity modification of chip after described substrate (4) and cover plate (2) micro Process arrange corresponding injection port, reaction chamber, buffer channel, Buffer Pool and gas outlet through surface treatment, rear bonding realizes airtight sealing-in.
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