CN105689026A - Separating preparation method of micro droplets and microarrays - Google Patents
Separating preparation method of micro droplets and microarrays Download PDFInfo
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- CN105689026A CN105689026A CN201410681689.9A CN201410681689A CN105689026A CN 105689026 A CN105689026 A CN 105689026A CN 201410681689 A CN201410681689 A CN 201410681689A CN 105689026 A CN105689026 A CN 105689026A
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Abstract
The invention belongs to the technical field of material interfacial wetting, and discloses a method used for preparing micro droplets and microarrays via separating single droplets. According to the method, droplets are dropped onto a patterned hydrophilic/hydrophobic substrate, contact force of the droplets with the patterned hydrophilic/hydrophobic substrate is controlled, and relative slip velocity between the droplets and the patterned hydrophilic/hydrophobic substrate is controlled, so that size accurate control separation of the droplets can be realized so as to obtain the micro droplets and microarrays. The method can be used for preparing a large amount of micro droplets from one drop of a sample solution, the size of obtained micro droplets is small, requirement on the sample solution is low, less sample is needed, separation efficiency is high, equipment is simple, operation is convenient, preparation cost is low, and application range is wider. The method possesses a promising application prospect in the fields of surface patterning, biological analysis and detecting, chemical reaction, and drug development.
Description
Technical field
The invention belongs to material interface imbibition technique field, particularly to separating the method that single drop prepares microlayer model and microarray。
Background technology
Microlayer model preparation is with a wide range of applications in bioanalysis and detection, chemical reaction, medicament research and development, patterned surface field。Current microlayer model preparation method mainly includes physics confinement (list of references: R.J.Jackman, D.C.Duffy, E.Ostuni, N.D.Willmore, G.M.Whitesides, Anal.Chem.1998,70,2280-2287；E.Ostuni, C.S.Chen, D.E.Ingber, G.M.Whitesides, Langmuir2001,17,2828-2834.), T-shaped duct microfluidic control standby (Patents number: CN104084247A, CN104107734A, CN104130932A), printing prepare (Patents number: CN104004652A, CN103974547A) etc.。But, there is a lot of defect in currently used method, big including required sample size, equipment needed thereby is costly or complicated, the microlayer model of preparation be in oil phase surround among be difficult to separate, and the microlayer model volume of preparation is big。Much more expensive for sample or amount and few in the case of, utilize current techniques to be difficulty with the separation of a large amount of microlayer model and the simple and easy preparation of microarray。
Summary of the invention
For solving above-mentioned the problems of the prior art, the present invention provides a kind of and utilizes surface differential to realize the method that microlayer model and microarray are prepared in drop cutting。
The cutting liquid dripping that the present invention proposes for the technical scheme of microlayer model and microarray is: by drop in the hydrophilic/hydrophobic substrate of patterning, by controlling the contact force between drop and substrate, and the Relative sliding speed between drop and substrate, the volume of drop can be carried out accurately control separation and obtain microlayer model and microarray。
Microlayer model separation method of the present invention is:
1) hydrophilic region and the water repellent region of patterning, is constructed at substrate surface；
2), drop is dragged with the pressure of 0-650 μ N and the speed of 1-20mm/s at substrate surface。
Described water repellent region contact angle is more than 100 °, it is preferable that 140-180 °；Described hydrophilic region contact angle is less than 60 °, it is preferable that 0-20 °。
Described drop is the liquid between polarity 28-80mN/m, and the volume of described drop is more than the resolution of pattern。
One or more in water, ethylene glycol, propylene glycol, glycerol, dimethyl sulfoxide of described drop, or selected from inorganic salt solution, nano-particle solution, cell solution, protein solution, DNA solution, blood plasma。
Described hydrophilic region is made up of hydrophilic material or hydrophilic base, or obtains through Hydrophilic modification；Described water repellent region is made up of hydrophobic material or hydrophobic substrate, or obtains through hydrophobically modified。
One or more in hydrophilic metal, hydrophilic metal oxide, glass, silicon chip, hydrophilic polymer film, hydrophilic timber, paper, hydrophilic fiber fabric of described hydrophilic base。
One or more in hydrophobic metals, hydrophobic metal oxide, hydrophobic polymer film of described hydrophobic substrate。
One or more in hydrophilic nanoparticles, hydrophilic micron particle, hydrophilic polymer, hydrophilic molecule of described hydrophilic material。
Described hydrophobic material selected from polydimethylsiloxane, polystyrene, politef, poly(perfluoropropene), hydrophobic nano particle, silane, silicon fluoride, chlorosilane, fluorosilicone, chlorine siloxanes, polrvinyl chloride, in one or more。
Described Hydrophilic modification method is one or more in uv-exposure, ion beam etching, Cement Composite Treated by Plasma, the process of Piranha washing liquid, spin-coating method, spraying process, electrospinning, printing, electrochemical corrosion。
Described hydrophobically modified method is that gas phase modifies method, liquid phase modifies one or more in method, spin-coating method, spraying process, electrospinning, printing, electrochemical corrosion。
After solvent volatilization in the above-mentioned microlayer model prepared, remaining solute forms microarray。
Above-mentioned microlayer model separation method bioanalysis and detection, chemical reaction, medicament research and development, sample separation field application。
Microlayer model of the present invention separates and microarray preparation method, a sample solution can be utilized to prepare substantial amounts of fine droplet, the microlayer model volume with separation is little, requiring low to sample solution, required sample is few, and separation efficiency is high, equipment is simple, the series of advantages such as easy and simple to handle, preparation cost is low, and range of application is wider。Drop separation of the present invention and microarray preparation method are with a wide range of applications at patterned surface, bioanalysis and detection, chemical reaction, Field of Drug Discovery。
Accompanying drawing explanation
Fig. 1. in the present invention, drop separation prepares the home-built equipment schematic diagram used by microlayer model array；
Fig. 2. the schematic diagram of microlayer model array prepared by drop separation of the present invention；
Accompanying drawing labelling: 1-X-Y mobile controller, 2-ten thousand/balance, 3-has the hydrophobic substrate of hydrophilic area of the pattern, and 4-prepares the original slip drop of microlayer model for drop cutting, the microlayer model of 5-drop cutting preparation。
Detailed description of the invention
Embodiment 1
An ethylene glycol solution is utilized to prepare ethylene glycol microlayer model array。It is cleaned silicon chip processing, specifically comprises the following steps that with acetone and alcoholic solution to silicon chip ultrasonic three times respectively, rinse well with deionized water。Then silicon chip is immersed in the toluene solution of the octadecyl trichlorosilane alkane that volume fraction is 1%, soak 10 minutes, dry up with nitrogen after taking-up。Whole process all carries out in clean room, it is prevented that dust in air etc. pollute silicon chip。Now the contact angle of silicon chip is 106 °。Then the preparation of hydrophilic region is carried out。Silicon chip is placed under photomask board, is then placed under the ultraviolet light that wavelength is 365nm, is exposed 10 minutes。In exposure process, being in the octadecyl trichlorosilane alkane molecular breakdown of transmission region, this region becomes hydrophilic region simultaneously。Fix an ethylene glycol solution with suction pipe, make ethylene glycol drop slide at the hydrophobicity silicon chip surface containing hydrophilic region with the speed of the pressure of 650 μ N and 10mm/s。Owing to hydrophilic region is different to the adhesiveness of ethylene glycol solution with water repellent region, the microlayer model array of ethylene glycol therefore can be obtained at hydrophilic region。
Embodiment 2
Dyestuff pattern is prepared with a dye solution。The filter paper that laboratory is conventional is carried out hydrophobization modification。Its experimental technique is: is placed in by filter paper in the vacuum desiccator of cleaning, places a piece of microscope slide wherein, drip perfluoro decyl trimethoxy silane on microscope slide。Then with oil pump to vacuum desiccator evacuation 10 minutes。Then exsiccator is placed 2 hours in 60 degrees Celsius of baking ovens。After taking-up, the static contact angle of filter paper is 140 °。The hydrophilic region preparation method of hydrophobicity filter paper is as follows: by 5 milliliters of water and 5 milliliters of ethylene glycol mix homogeneously, be added thereto to the PAA (molecular weight 3000) that mass fraction is 1%。After stirring, as the ink of inkjet printing。Printing on hydrophobicity filter paper with ink-jet printer, in printing, the region of ink becomes hydrophilic region, it does not have in printing, the region of ink remains as water repellent region。A droplet dye solution dropper is fixed, patterns its surface with the speed of 0 pressure and 20mm/s in hydrophobe and roll。Owing to hydrophilic region is high to the adhesiveness of dye solution, water repellent region is low to the adhesiveness of dye solution, and adhering difference makes dye solution be cut into microlayer model。The hydrophilic region of its surface is prepared into pattern, then can obtain the microlayer model pattern of dye solution。After solvent is evaporated completely, filter paper just obtains dyestuff pattern。
Embodiment 3
The preparation of sodium chloride crystal。Prepared by hydrophilic base: by ultrasonic to microscope slide ethanol and acetone three times, then clean with distilled water flushing, then dries up with nitrogen。Whole process carries out in clean room, it is ensured that glass sheet surface is not by the impurity pollution in air。Then by micro-machined method prepared by surface the pattern of hydrophobicity Teflon。Pattern thickness is between 50 nanometers to 10 microns。Now, the surface being coated with by Teflon is water repellent region, it does not have be hydrophilic region by the surface coated region of Teflon。Being fixed by a droplet saturated nacl aqueous solution dropper, the Teflon that the pressure of 200 μ N and the speed of 1mm/s pattern in hydrophobe/hydrophilic glass surface slides。Teflon surface is little to saturated nacl aqueous solution adhesion, and hydrophilic glass sheet is big to saturated nacl aqueous solution adhesion。Saturated nacl aqueous solution is cut in the hydrophilic glass panel region that adhesion is big, becomes microlayer model。Along with evaporation of water in the microlayer model under cutting, sodium chloride gradually forms nucleus and starts to grow up to crystal。This kind of method prepares sodium chloride crystal efficiency height, and the time is short, easily operates。
Embodiment 4
The controllable ration of microsphere separates。Prepared by hydrophobe patterned substrate: being cleaned silicon chip processing, concrete operations are as follows: with acetone and alcoholic solution to silicon chip ultrasonic three times respectively, then cleans with distilled water, and dries up with nitrogen。Then the positive glue of spin coating one layer photoetching glue on silicon chip, the positive glue thickness of photoresist is between 200 nanometers to 3 microns。Under photomask board is protected, carry out selecting uv-exposure to photoresist with ultraviolet light。The region being mapped to by ultraviolet lighting can developed dose wash, it does not have still there is photoresist in the region being mapped to by ultraviolet lighting。Processing after completely with developing agent, carry out hydrophobically modified by the method for vapour deposition to silicon chip, concrete operation method is with example 2。After modification completes, part is covered ultrasonic 1 hour of the silicon chip acetone of photoresist, remove remaining photoresist。Now, the region of new removing photoresist, owing to not being modified in hydrophobically modified process, is therefore hydrophilic region。The wafer sections exposed in hydrophobically modified process becomes water repellent region。Then the silicon chip surface patterned in hydrophobe with the pressure of 500 μ N and the speed of 15mm/s with the polystyrene microsphere solution (mass fraction 5%) that dropper droplet diameter of control is 5 microns slides。Owing to hydrophilic and hydrophobic region is different to the adhesion of microspheres solution, therefore microspheres solution drop is cut, and becomes microlayer model。After solvent is evaporated in microlayer model, then can realize the separation to microsphere。The speed slided at silicon chip surface by the size of capping hydrophilic region and microspheres solution drop and pressure, it is possible to achieve the quantity isolating microsphere is controlled, thus realizing the separation of the controlled quantity of microsphere。
Embodiment 5
Unicellular separation。The preparation of hydrophobe patterned substrate: by butyl methacrylate, dimethacrylate time ethyl ester, 1-decanol, Hexalin, 2,2-dimethoxy-2-phenyl acetophenone 10:16:24:49:1 in mass ratio ratio mix homogeneously。Then mixture is placed between two parallel glass sheets that distance is 1mm, adds photomask board, then irradiate 10 minutes under 365nm ultraviolet light, carry out selectivity light initiation polymerization。Removing sheet glass after polymerization, the polymeric film of formation is hydrophilic and hydrophobic patterned film。On photomask board, photopolymerization can be caused in the region of printing opacity, and after the polymer polymerizing below this region, contact angle is 155 °。On photomask board, photopolymerization can not be caused in other lighttight regions, and the polymeric film contact angle below this region is 20 °。Fix a Cell sap with dropper, slide with the substrate surface that the speed of the pressure of 300 μ N and 5mm/s patterns in hydrophobe。Owing to hydrophilic region is different to the adhesiveness of Cell sap with water repellent region, therefore Cell sap drop can be cut into the microlayer model containing cell。The size of concentration and hydrophilic region by controlling Cell sap, it is possible to achieve single celled separation。
Claims (8)
1. a microlayer model separation method, it is characterised in that its concrete operation step is:
1) hydrophilic region and the water repellent region of patterning is constructed at substrate surface；
2) drop is dragged with the pressure of 0-650 μ N and the speed of 1-20mm/s at substrate surface。
2. microlayer model separation method according to claim 1, it is characterised in that described water repellent region contact angle is more than 100 °, it is preferable that 140-180 °；Described hydrophilic region contact angle is less than 60 °, it is preferable that 0-20 °。
3. microlayer model separation method according to claim 1, it is characterised in that described drop is the liquid between polarity 28-80mN/m, and the volume of described drop is more than the resolution of pattern。
4. microlayer model separation method according to claim 1, it is characterized in that, one or more in water, ethylene glycol, propylene glycol, glycerol, dimethyl sulfoxide of described drop, or selected from inorganic salt solution, nano-particle solution, cell solution, protein solution, DNA solution, blood plasma。
5. microlayer model separation method according to claim 1, it is characterised in that described hydrophilic region is made up of hydrophilic material or hydrophilic base, or obtains through Hydrophilic modification；Described water repellent region is made up of hydrophobic material or hydrophobic substrate, or obtains through hydrophobically modified。
6. microlayer model separation method according to claim 5, it is characterized in that, one or more in hydrophilic metal, hydrophilic metal oxide, glass, silicon chip, hydrophilic polymer film, hydrophilic timber, paper, hydrophilic fiber fabric of described hydrophilic base；
One or more in hydrophobic metals, hydrophobic metal oxide, hydrophobic polymer film of described hydrophobic substrate；
One or more in hydrophilic nanoparticles, hydrophilic micron particle, hydrophilic polymer, hydrophilic molecule of described hydrophilic material；
Described hydrophobic material selected from polydimethylsiloxane, polystyrene, politef, poly(perfluoropropene), hydrophobic nano particle, silane, silicon fluoride, chlorosilane, fluorosilicone, chlorine siloxanes, polrvinyl chloride, in one or more；
Described Hydrophilic modification method is one or more in uv-exposure, ion beam etching, Cement Composite Treated by Plasma, the process of Piranha washing liquid, spin-coating method, spraying process, electrospinning, printing, electrochemical corrosion；
Described hydrophobically modified method is that gas phase modifies method, liquid phase modifies one or more in method, spin-coating method, spraying process, electrospinning, printing, electrochemical corrosion。
7., after the solvent volatilization in the microlayer model prepared according to the arbitrary described microlayer model separation method of claim 1-6, remaining solute forms microarray。
8. microlayer model separation method according to claim 1 bioanalysis and detection, chemical reaction, medicament research and development, sample separation field application。
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| CN110082063A (en) * | 2019-03-04 | 2019-08-02 | 中国科学院化学研究所 | The method and purposes of rotary motion after a kind of control droplet collision |
| CN110465337A (en) * | 2018-05-11 | 2019-11-19 | 中国石油化工股份有限公司 | Chip and its liquid stream control method and application |
| CN111139517A (en) * | 2020-01-03 | 2020-05-12 | 浙江工业大学 | Preparation method and device of array type liquid drop crystal |
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| CN112275332A (en) * | 2020-09-17 | 2021-01-29 | 厦门大学 | Self-powered digital microfluidic chip and droplet control method |
| CN113073029A (en) * | 2021-03-17 | 2021-07-06 | 长春长光辰英生物科学仪器有限公司 | Infiltration modified cell sorting chip for laser induced transfer and sorting method |
| CN114471397A (en) * | 2021-12-22 | 2022-05-13 | 天津大学 | Nucleic acid synthesis-oriented microreactor array chip and preparation method thereof |
| CN114471397B (en) * | 2021-12-22 | 2024-04-02 | 天津大学 | Micro-reactor array chip for nucleic acid synthesis and preparation method thereof |
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