CN109701671A - A kind of microlayer model array chip and production and preparation method thereof - Google Patents
A kind of microlayer model array chip and production and preparation method thereof Download PDFInfo
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- CN109701671A CN109701671A CN201811611242.9A CN201811611242A CN109701671A CN 109701671 A CN109701671 A CN 109701671A CN 201811611242 A CN201811611242 A CN 201811611242A CN 109701671 A CN109701671 A CN 109701671A
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- microlayer model
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Abstract
The invention discloses a kind of microlayer model array chips and production and preparation method thereof, include: substrate；The microwell array being made of several micropores is provided in the substrate, which has 3D shape, and pore openings are gradually big, and micropore side wall at least one side is inclined at an angle, and the contact angle of the substrate and liquid is greater than 80 °.The present invention can be conducive to that gas in micropore excludes and microlayer model leaves using pore openings gradually big technology, guarantee the formation efficiency of microlayer model, the microlayer model of generation is small in size, stability is high, small with base material contact area.Manufacturing method is simple, at low cost.Application method is simply various, low to external equipment and operator's technical requirements, and adaptation range is extensive.
Description
Technical field
The present invention relates to a kind of microfluidic fields, and in particular to a kind of chip for generating microlayer model array and its manufacture and makes
With method, apply in fields such as analysis tests.
Background technique
Microlayer model array in fields such as biomolecular science, chemical analysis and drug screenings using very extensive, efficiently
Microlayer model array is particularly significant for high-throughput test analysis.
There is also many technical problems, such as drop formation of micro-/nanoliter volumes for efficient microlayer model array technique；A large amount of batches
Array is measured to generate；Simply, it quickly generates and stability etc..
Currently, many microlayer model production methods: 1, the device is complicated for control, generally requires lasting external force, such as pump pressure, vacuum
Negative pressure etc.；2, micro-fluidic chip generally requires the composite construction of multilayer for constituting closed cavity；3, microlayer model and substrate material
Expect that contact area is big, volume is uneven, and position is inaccurate etc.；4, usage mode is complex, and the time is long；5, chip needs chemistry
A possibility that handling, increasing pollution effect drop characteristics.
(R.J.Jackman, D.C.Duffy, E.Ostuni, the N.D.Willmore and such as Jackman
G.M.Whitesides, Anal.Chem.1998,70,2280.) when generating droplet array, in such a way that painting is scraped
It can not solve the problems, such as that liquid of the contact angle greater than 80 ° or more generates droplet array, the gas in micropore can not be successfully discharge,
Must by big quantity of fluid carry out lateral dipping in the way of could generate droplet.This mode causes the wave for being filled liquid
Take and pollutes.
The droplet that Yasuga etc. (WO2018/003856 A1) is generated generates, the property of oily phase by the shearing of oily phase
The uniformity of drop is affected, device work limits large area droplet array dependent on the interface channel between micropore
It generates and its applies.
Complexity is constituted, using time-consuming, the deficiency of microlayer model controllability difference for system in the generation of microlayer model array, urgently
Wait need, a kind of microlayer model array is raw quickly, droplets stable is accurate, using simple generation method.
Summary of the invention
In order to solve prior art problem, it is an object of the present invention to overcome the deficiencies of the prior art, and to provide one kind
Microlayer model array chip and production and preparation method thereof, by changing micropore on micro-fluidic chip (i.e. microlayer model array chip)
Shape keeps its opening gradually big, so that microfluid is easier to enter, microfluid is trapped in micro- under the contact action with base material
In hole, even if under external force, liquid to be filled flows through microwell array, micro- liquid can be also retained down in micropore, then on surface
Under the smaller oily phase separation of tension, microlayer model array is formed.
In order to achieve the above object, the present invention provides a kind of microlayer model array chip, which includes: substrate；The base
The microwell array being made of several micropores is provided on bottom, the micropore has 3D shape, and pore openings are gradually big, micropore
Side wall at least one side is obliquely installed, and the contact angle of the substrate and drop is greater than 80 °.
Preferably, the 3D shape of the micropore is prismatoid, appointing in pyramid, hemisphere, semiellipsoid, drops
The combination for one or more of anticipating.
Preferably, the volume of each micropore is same or different.
Preferably, the microwell array shape selects rectangle, triangle, trapezoidal or irregular shape.
Preferably, the base material and liquid to be filled meet liquid slide angle greater than 10 °, Liquid contact angle is big
In 80 °.It is preferred that using PDMS base material, liquid to be filled is usually aqueous solution.
The present invention also provides a kind of preparation methods of above-mentioned microlayer model array chip, using bionical reverse mould method or light
It carves reverse mould method and prepares microlayer model array chip.
Preferably, the photoetching reverse mould method includes:
Step 1, lithography process prepares several grooves, forms the mold with groove array；
Step 2, the solution of base material is poured into the groove of mold, and carries out heating cure, form basal layer；
Step 3, the basal layer after solidifying is removed, with groove；
Step 4, in the groove of the basal layer by the solution spin coating of the base material after dilution after hardening；
Step 5, heating cure forms the microlayer model array chip with several microwell arrays.
Preferably, the bionical reverse mould method includes:
Step a) takes new new lotus leaf to clean, drying；
Step b), lotus leaf is fixed；
Step c) pours into the solution of base material, and carries out heating cure, forms basal layer；
Step d) takes out the basal layer after solidifying.
The present invention also provides a kind of application methods of above-mentioned microlayer model array chip, and this method includes:
Liquid to be filled is added to any one or more micropores of microlayer model array chip in S1；
S2, under external force traction, liquid to be filled flows through microwell array through substrate, is evenly distributed to microlayer model array chip
Micropore in；
S3 is added dropwise oil droplet and is rinsed under external force traction to substrate, liquid to be filled remaining in substrate is washed down,
Oil droplet dispersed encapsulated liquid to be filled is simultaneously distributed to microwell array, and liquid in micropore is made to become a sphere, with greatly reduce with
The long-time of micro-pore wall contacts, and avoids pollution；
S4 seals microlayer model array chip using coverslip.
In S3, the purpose that oil droplet is added is as follows: 1, to stay in the liquid in micropore and form drop spherical shape；2, drop is prevented
Volatilization；3, substrate surface residual liquid is washed out；4, from environmental pollution.
Preferably, the external force traction includes the group of any one or more in mechanical external force, gravity, centrifugal force
It closes.
Technical concept of the invention are as follows: the effect at sluggish angle is utilized, so that miniflow in design pore openings gradually big form
Body is trapped in micropore the extensive generation, it can be achieved that batch microlayer model under the contact action with base material, adds oil
Phase, further such that microlayer model and base material contact area reduce, it is small on being influenced caused by micro- liquid biochemical reaction and analysis.
Micro-fluidic chip is used for using the production of the micro-processing technologies such as bionical reverse mould, photoetching reverse mould, is generated for microlayer model array, microlayer model
Stability is high, and micro-fluidic chip is at low cost.
The effect at the sluggish angle is similar to a drop and drips in one plane, when plane tilts very big angle ability
Slide drop.The sluggish angle of the intrinsic property of PDMS layer itself is bigger, in addition opening is gradually big, it is more with drop contact area,
Sluggish angle effect is more obvious, and liquid once enters to be difficult to skid off.
The present invention compared with prior art, has the following obvious advantages:
1. microwell array can be different using shape, size, array manner in micro-fluidic chip of the present invention, micro- liquid of generation
More application scenarios can be used in size, the array manner of drop.
2. microwell array pore openings are gradually big in micro-fluidic chip of the present invention, it is ensured that liquid to be filled can smoothly enter into micro-
Kong Zhong, to generate micro- liquid array.This kind of mode produces that microlayer model is high-efficient, and microlayer model is uniform and stable.
3. the microlayer model shape that micro-fluidic chip of the present invention is formed is close to spheroidal, small with base material contact area,
Influence of the base material property to micro- liquid component is reduced, the performance of biochemical test analysis is improved.
4. being used for micro-fluidic chip using the production of the micro-processing technologies such as bionical reverse mould, photoetching reverse mould, it to be used for microlayer model array
It generates.
5. application method of the present invention is simple and efficient, liquid to be filled can be driven using mechanical external force, gravity, centrifugal force etc.
The mode of microwell array is flowed through, it is not only easy to operate, but also can be used in different application scene.
Detailed description of the invention
Fig. 1 is the pore openings gradually big structure schematic diagram of microlayer model array chip of the invention.
Fig. 2 is the bionical reverse mould manufacturing method of microlayer model array chip of the invention.
Fig. 3 shows for the drop of the lotus leaf reverse mould PDMS of the invention microlayer model array chip formed, statistics, pattern, section
It is intended to.
Fig. 4 a-4f is the photoetching reverse mould manufacturing method of microlayer model array chip of the invention.
Fig. 5 is drop, statistics, pattern, section of the present invention using the microlayer model array chip of photoetching reverse mould technology preparation
Schematic diagram.
Fig. 6 is microlayer model array chip application method schematic diagram of the invention.
Specific embodiment
Below in conjunction with drawings and examples, the following further describes the technical solution of the present invention.
" contact angle " as described herein (contact angle) refers to the solution-air made by gas, liquid, solid three-phase point of intersection
The tangent line at interface, this tangent line are the measurements of wetness degree in the angle theta between solid-liquid boundary line of one side of liquid.
" slide angle " i.e. as described herein roll angle, when referring to that drop just rolls on the sloped surface, nauropemeter
Face and horizontal plane are formed by critical angle, are indicated with α, are the important methods for characterizing a particular surface wetability.
The slide angle of base material of the present invention and liquid to be filled is greater than 10 °, and contact angle is greater than 80 °, may be selected
PDMS (Polydimethylsiloxane, dimethyl silicone polymer), POE (Polyolefin elastomer, polyolefin elastic
Body), EPDM (Ethylene Propylene Diene Monomer, ethylene propylene diene rubber), PMMA
(Polymethylmethacrylate, polymethyl methacrylate), and PTFE (Poly tetra fluoroethylene, poly- four
Vinyl fluoride), any one or any two or more composite materials in PR (Polystyrene, polystyrene).
" opening is gradually big " as described herein refers to that the bore of the opening of micropore is gradually increased from bottom to top.
As shown in Figure 1, micro-fluidic chip is the base material that the microwell array being made of several micropores 1 is distributed with, micropore 1
Opening it is gradually big, micropore side wall at least one side is inclined at an angle.
The micropore of micro-fluidic chip can be different 3D shape, and the 3D shape of micropore is including but not limited to trapezoidal
Any one in body, pyramid, hemisphere, semiellipsoid, drops etc., the side wall at least one side of micropore favours substrate material
Material.
In some preferably embodiments, the microwell array shape of micro-fluidic chip can be different flat shape, including
But be not limited to rectangle, triangle, trapezoidal or irregular shape etc..
As shown in Fig. 2, the base material of micro-fluidic chip can use bionical reverse mould technology comprising the steps of: a) take
One piece of fresh lotus leaf cleans impurity, drying moisture；B) lotus leaf (face-up) is fixed on culture dish by double-sided adhesive；
C) PDMS containing premixed liquid is poured into, and carries out heating cure；D) PDMS after solidifying is taken out, there is similar lotus leaf surface
Microwell array.Since there are many minute protrusions structures in lotus leaf front, the surface PDMS has reformed into small bowl configurations after reverse mould.
The drop of microlayer model array chip that the lotus leaf reverse mould PDMS is formed, statistics, pattern, section signal as shown in figure 3,
Wherein, a is microwell array part shape appearance figure, and b is the volume distribution statistics figure of 2297 drops, and c is the micropore shape of partial enlargement
Shape, d are micropore schematic cross-section.
As shown in Fig. 4 a-4f, microlayer model array chip can be prepared using photoetching reverse mould technology:
Step 1, lithography process SU-8 substrate 10 prepares several grooves 11, as shown in fig. 4 a.
Step 2, the PDMS containing premixed liquid is poured into the groove of SU-8 substrate, and carries out heating cure, form PDMS
Layer 20, as shown in Figure 4 b.
Step 3, the PDMS layer 20 after solidifying is removed, with groove 21, as illustrated in fig. 4 c.
Step 4, by the groove 21 of the PDMS layer 20 of the PDMS spin coating containing premixed liquid after hardening after dilution, such as scheme
Shown in 4d；
Step 5, heating cure, during heating, diluent (n-hexane, toluene etc.) are volatilized (see Fig. 4 e), and formation is opened
The micropore 1 of mouth gradually big shape, forms the micro-fluidic chip with several microwell arrays, as shown in fig. 4f.
Illustrated using the drop of the microlayer model array chip of photoetching reverse mould technology preparation, statistics, pattern, section such as Fig. 5 institute
Show, wherein a is microwell array part shape appearance figure, and b is the volume distribution statistics figure of 576 drops, and c is the micropore of partial enlargement
Shape, d are micropore schematic cross-section.
The application method of micro-fluidic chip provided by the invention as shown in fig. 6, comprising:
Reagent is added into micro-fluidic chip by S1；
S2 is exerted a force using coverslip or scraper or scraping blade in micro-fluidic chip upper surface level, under external force traction, wait fill out
Filling liquid body flows through microwell array with certain speed, so that reagent is evenly distributed in the microwell array of micro-fluidic chip；Described
External force refers to be driven using mechanical external force, gravity, centrifugal force etc.；
S3 is added oil droplet, is rinsed, and acts on as follows: 1, making the liquid stayed in micropore form drop spherical；2, prevent
Only drop volatilizees；3, substrate surface residual liquid is washed out；4, protect drop from environmental pollution.
S4 covers the microwell array of sealing micro-fluidic chip using coverslip, it is ensured that drop is fixed on the inside, from outer
The pollution of boundary's impurity.
In conclusion the base material intrinsic property sluggishness angle that microlayer model array chip of the invention uses is bigger, add
Upper pore openings are gradually big, more with drop contact area, and sluggish angle effect is more obvious, liquid once enter difficulty skid off, it can be achieved that
The extensive generation of batch microlayer model adds oily phase, and microlayer model and base material contact area are small, to micro- liquid biochemical reaction
It is small with influence caused by analysis.The present invention has the advantages that chip structure is simple, microlayer model formation efficiency is high, easy to operate.
It is discussed in detail although the contents of the present invention have passed through above preferred embodiment, but it should be appreciated that above-mentioned
Description is not considered as limitation of the present invention.After those skilled in the art have read above content, for of the invention
A variety of modifications and substitutions all will be apparent.Therefore, protection scope of the present invention should be limited to the appended claims.
Claims (10)
1. a kind of microlayer model array chip, which is characterized in that the chip includes: substrate；It is provided in the substrate by several micropores
The microwell array of composition, the micropore has 3D shape, and pore openings are gradually big, and the inclination of micropore side wall at least one side is set
It sets, the contact angle of the substrate and drop is greater than 80 °.
2. microlayer model array chip as described in claim 1, which is characterized in that the 3D shape of the micropore be prismatoid,
Pyramid, hemisphere, semiellipsoid, any one or the combination of several of them in drops.
3. microlayer model array chip as described in claim 1, which is characterized in that the volume of each micropore is same or different.
4. microlayer model array chip as described in claim 1, which is characterized in that the microwell array shape selects rectangle, three
Angular, trapezoidal or irregular shape.
5. microlayer model array chip as described in claim 1, which is characterized in that the slide angle of the base material and drop
Greater than 10 °.
6. a kind of preparation method of the microlayer model array chip as described in any one of claim 1-5, which is characterized in that adopt
Microlayer model array chip is prepared with bionical reverse mould method or photoetching reverse mould method.
7. the preparation method of microlayer model array chip as claimed in claim 6, which is characterized in that the photoetching reverse mould method packet
Contain:
Step 1, lithography process prepares several grooves, forms the mold with groove array；
Step 2, base material solution is poured into the groove of mold, and carries out heating cure, form basal layer；
Step 3, the basal layer after solidifying is removed, with groove；
It step 4, will be in the groove of the basal layer of base material solution spin coating after hardening after dilution；
Step 5, heating cure forms the microlayer model array chip with several microwell arrays.
8. the preparation method of microlayer model array chip as claimed in claim 6, which is characterized in that the bionical reverse mould method packet
Contain:
Step a) takes new new lotus leaf to clean, drying；
Step b), lotus leaf is fixed；
Step c) pours into base material solution, and carries out heating cure, forms basal layer；
Step d) takes out the basal layer after solidifying.
9. a kind of application method of the microlayer model array chip as described in any one of claim 1-5, which is characterized in that should
Method includes:
Solution to be filled is added to any one or more micropores of microlayer model array chip in S1；
S2, under external force traction, liquid to be filled flows through microwell array through substrate, is evenly distributed to the micro- of microlayer model array chip
Kong Zhong；
S3 is added dropwise oil droplet and is rinsed under external force traction to substrate；
S4 seals microlayer model array chip using coverslip.
10. the application method of microlayer model array chip as claimed in claim 9, which is characterized in that the external force traction packet
Containing the combination of any one or more in mechanical external force, gravity, centrifugal force.
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Cited By (6)
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| CN110643483A (en) * | 2019-09-06 | 2020-01-03 | 上海交通大学 | Method for generating liquid drop array on microfluidic chip |
| CN110653015A (en) * | 2019-10-30 | 2020-01-07 | 潍坊医学院 | Microbial detection chip with high sample filling rate and filling method thereof |
| CN111330660A (en) * | 2020-03-10 | 2020-06-26 | 中国科学院苏州生物医学工程技术研究所 | Centrifugal high-flux micro-droplet preparation chip |
| CN112816706A (en) * | 2021-01-06 | 2021-05-18 | 上海理工大学 | Digital ELISA system and using method thereof |
| CN113083386A (en) * | 2021-04-02 | 2021-07-09 | 重庆大学 | Simple and rapid liquid sample discretization chip and using method thereof |
| CN113477285A (en) * | 2021-07-21 | 2021-10-08 | 杜林� | Micro-droplet array chip system and method |
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| CN113083386A (en) * | 2021-04-02 | 2021-07-09 | 重庆大学 | Simple and rapid liquid sample discretization chip and using method thereof |
| CN113477285A (en) * | 2021-07-21 | 2021-10-08 | 杜林� | Micro-droplet array chip system and method |
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