CN1302282C - Method for preparing noumenal modification polymethyl methacrylate micro flow control chip - Google Patents
Method for preparing noumenal modification polymethyl methacrylate micro flow control chip Download PDFInfo
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- CN1302282C CN1302282C CNB2004100934830A CN200410093483A CN1302282C CN 1302282 C CN1302282 C CN 1302282C CN B2004100934830 A CNB2004100934830 A CN B2004100934830A CN 200410093483 A CN200410093483 A CN 200410093483A CN 1302282 C CN1302282 C CN 1302282C
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Abstract
The present invention belongs to the technical field of biological detection, more specifically a preparation method for bulk-modified polymethyl methacrylate micro-fluidic chips. Methyl methacrylate and modifiers are mixed, a small amount of thermal initiators and photoinitiator are dissolved in the mixed solution, a monomer solution is prepolymerized by being heated in a water bath, the prepolymerized solution is injected in a chip mold, bulk polymerization is initiated by the irradiation of ultraviolet light, substrates of micro-fluidic chips are obtained through manufacture, and the obtained micro-fluidic chips are encapsulated by cover membranes. The present invention introduces functional modifiers, such as methacrylic acid or p-vinylpyridine, etc. in the main chain of polymethylmethacrylate, regulates the charge property of the surfaces of monomers, and controls electroosmotic flows from the molecular level of chip materials. Thereby, separation is improved, the performance of chips is improved, and the application of chips is widened. The chips have the advantages of simple manufacture technology, low cost and mass production, and has favorable application prospects in the fields of environmental monitoring, clinical diagnosis, life science research, food analysis, industrial on-line analysis, etc.
Description
Technical field
The invention belongs to technical field of biological, be specifically related to a kind of noumenal modification polymethyl methacrylate micro flow control chip preparation method.
Background technology
Since [1] such as nineteen ninety A.Manz has proposed micro-full analytical system (since the μ-TAS) first, as a frontier interdisciplinary, its target is to handle whole microminiaturized, the integrated and portability that detects by micro electronmechanical processing (MEMS) technology and biotechnology realization chemical analysis system from sample, is the important directions and the forward position of present analytical instrument development.Micro-fluidic chip is an architectural feature with the microchannel network then, it is the emphasis of current micro-total analysis system development, and efficient with it, fast, few, the low consumption of reagent dosage and integrated level advantages of higher caused domestic and international analysis and life science circle relevant expert's extensive concern, shown good prospects for application in fields such as environmental monitoring, clinical diagnosis, Pharmaceutical Analysis, legal medical expert and military affairs, various new micro-fluidic chip preparations and detection technique emerge in an endless stream.
Micro-fluidic chip mainly uses glass and polymer chip [2], and the glass-chip process technology requires high, needs specialized apparatus, is difficult to adopt mould to be produced in enormous quantities, and the price comparison costliness has limited its application.So polymer chip is developed, wherein polymethylmethacrylate (PMMA) and dimethyl silicone polymer (PDMS) are two kinds of polymkeric substance commonly used [3].Technology such as injection moulding, die and casting are mainly adopted in the making of polymer chip, but the capillary slot on the plastic chip of making has metaboly, with design load certain difference is arranged, and the reappearance of chip chamber of the same race is not good.The monomer injection molding is directly produced the polymer core chip technology, to there be the silicon chip or the metal anode membrane of chip microfluxion to make mould, again polymer monomer such as methyl methacrylate etc. is injected wherein initiated polymerization, can be made into chip after the demoulding, have that method is easy, raw materials cost is cheap, high repeatability and other advantages is fit to produce in enormous quantities.Polymer chip life-span and stability are poor than glass, but can change its surface physics and chemical property by surface and noumenal modification, improve separation, example enrichment and purpose such as derive to reach.The chemical modification of polymer chip at present still is in the starting stage, relevant research less [3], but can predict, this will be an important research direction.
List of references
[1]Manz?A，Graber?N，Widmer?HM.Sens.Actuators?B?1990，1，244-248.
[2]Verpoorte?E.Electrophoesis?2002，23，677-712.
[3]Becker?H，Locascio，LE.Talanta?2002，56，267-287.
Summary of the invention
The objective of the invention is to propose a kind of preparation method of noumenal modification polymethyl methacrylate micro flow control chip, the charge on regulation and control polymethylmethacrylate surface is controlled electroosmotic flow and is improved chip performance from the molecular level of chip material.
The noumenal modification polymethyl methacrylate micro flow control chip preparation method that the present invention proposes, be that methyl methacrylate is mixed with dressing agent (as methacrylic acid or to vinylpyridine etc.), then thermal initiator azoisobutyronitrile and light trigger styrax are dissolved in this mixed solution, heating is 15-20 minute in 80-90 ℃ of water-bath, makes monomer solution pre-polymerization (becoming glycerine shape clear solution); Above-mentioned pre-gathering solutions is injected by containing micro-fluidic chip protrusion microstructure silicon chip formpiston and two chip moulds that thin glass plate constitutes, shone pre-gathering solutions 30-60 minute with ultraviolet light by the mould glass, cause bulk polymerization, make the micro-fluidic chip substrate.
The advantage that the present invention utilizes bulk polymerization to make, functional dressing agent (as methacrylic acid or to vinylpyridine etc.) is introduced the main chain of polymethylmethacrylate, charge to the polymethylmethacrylate surface is regulated and control, from the molecular level of chip material control electroosmotic flow, separate, improve chip performance and widen the purposes of chip thereby improve.
Among the present invention, adopt computer aided design software design microfluidic chip structure, typical design as shown in Figure 1.Constitute by single right-angled intersection microchannel and solution connection holes, adopt high resolving power (as 3600dpi) laser photocomposing system on transparent membrane, to make the mask negative film, microchannel width on the mask is 40-100 μ m, solution connection holes diameter 1-3mm, wherein separate microchannel 2 and sample intake passage 7 and solution hole 1,4,5 and 6 (Fig. 1) for transparent, remainder is a black.The erect image of design drawing is seen Fig. 1.In silicon chip (p type through oxidation processes, thick 500 μ m, 4 inches of diameters, crystal orientation＜100 〉, surface silica dioxide oxidation bed thickness 100nm) applies one deck negative photoresist (as the SU-8 photoresist) by the spin-coating technology, cover mask (the chip capillary microfluxion that contains design) then, after ultraviolet exposure and baking processing, with supporting developer immersion treatment, flush away is not exposed the photoresist layer of part (kapillary and solution hole beyond zone) in acetone and isopropyl alcohol respectively, toast in baking oven then, the photoresist sclerosis that capillary channel and solution connection holes are partly exposed is in rare HF-NH
4F solution etching off silicon chip surface is not exposed the SiO of part
2Layer, then in 50-70 ℃ with about 1 hour of the exposed silicon chip of 40%KOH aqueous solution (containing 5% isopropyl alcohol) etching, promptly making silicon chip formpiston 12,10 be the chip size cavity, 11 is open slot, 13 microchannels for silicon male mold surfaces protrusion.Silicon chip one side and two blocks of sheet glass 8 and 8 that microchannel will be arranged, clamping a middle hollow out is the aluminium sheet 9 (about 2mm is thick) of the rectangle of chip size, constitutes chip mould 14 (Fig. 2).
Among the above-mentioned preparation method, the consumption of dressing agent is generally the 1-3% of methyl methacrylate monomer weight, the consumption of thermal initiator and light trigger is respectively the 0.1-0.2% of monomer weight, attention will prevent that sealing enters in pre-collecting process, avoid temperature too high simultaneously, otherwise can be sudden and violent poly-because of causing, cause the waste of material.Pre-polymerization later stage polymerization speed is accelerated, and polymerization should be controlled in 20 minutes, otherwise the polymeric solution viscosity of pre-polymerization is excessive, is difficult for pouring into mould and has easily introduced bubble.
During preparation, above-mentioned pre-gathering solutions is injected mould 14 by mould openings groove 11, open slot 11 caused bulk polymerization with 20W uviol lamp (365nm is apart from 4-5 centimetre) in 30-60 minute by mould glass irradiation pre-gathering solutions up.Simultaneously respective liquid potpourri injection molding is about polymerization between the sheet glass of 80-120 μ m in the slit, obtains the epiphragma 17 of same material after the demoulding.After treating the polymeric solution sclerosis of pre-polymerization, with mould demoulding in ultrasonic 10 minutes in 30-50 ℃ of water-bath.The micro-fluidic chip substrate 15 channel ends borings ( solution connection holes 1,4,5 and 6 is seen Fig. 1, aperture 2mm) of the demoulding are used to connect solution.For prevent that microchannel from stopping up in encapsulation process, before the overlay film, in the microchannel of substrate, fill low-melting water-soluble polymers polyglycol (PEG) 1500 by knife coating, fusing point is 41～46 ℃, because of the low fusing easily of fusing point, can under liquid state, be filled in the microchannel, behind the cooling curing, use wet filter paper wiping micro-fluidic chip substrate surface to remove the outer PEG of passage.With chloroform epiphragma 17 is bonded on the micro flow chip substrate 15 again, in 70-80 ℃ of water-bath, the microchannel interpolymer is melted then, by the hot water in the syringe it being released microchannel is also cleaned outward, promptly can be made into noumenal modification polymethyl methacrylate micro flow control chip finished product 18, see Fig. 4.The bonding encapsulation polymethyl methacrylate micro flow control chip of this low melting point water-soluble polymers secondary solvent method belongs to initiative, what widely use for a long time is packaging by hot pressing, because difficult control of temperature, passage is easily stifled when packaging by hot pressing easily, and this method significantly can reduce the rejection rate of micro-fluidic chip preparation.
The noumenal modification polymethyl methacrylate micro flow control chip that the present invention makes is easy and simple to handle, favorable reproducibility, highly sensitive, the range of linearity is wide, amount of samples is few, wherein the methacrylic acid noumenal modification polymethyl methacrylate micro flow control chip can be used for the compartment analysis of phenols environmental contaminants, the analysis efficiency height.Concrete visible following test experiments result:
Use methacrylic acid noumenal modification polymethyl methacrylate micro flow control chip shown in Figure 1,50 μ M2 of acquisition, the electrophoresis pattern of the electrophoretogram of 6-xylenol, phenol and parachlorophenol is seen Fig. 5.Test condition is: separation voltage is+2000V, sample introduction voltage is+2000V, and sample injection time is 3s, and buffer solution is 50mM borate buffer solution (pH 9.2), detecting electrode is the carbon disk electrode of diameter 200 μ m, and the detection current potential is 0.95V (with respect to the Ag/AgCl electrode).The range of linearity is 0.1 μ mol/L-750 μ mol/L, is limited to 0.08-0.1 μ mol/L under detecting.Measure 50 μ M2 10 times, the relative standard deviation of 6-xylenol, phenol and parachlorophenol peak-to-peak signal is respectively 3.6%, 2.9% and 4.1%, show that this noumenal modification polymethyl methacrylate micro flow control chip range of linearity is wide and reappearance is good, fast efficient, in 100 seconds, just can separate and detect simultaneously three kinds of phenolic comp ' ds pollution fully, can be used for the mensuration of actual sample.
Description of drawings
Fig. 1 is the typical micro-fluidic chip design drawing that the present invention relates to.
Fig. 2 is the structural drawing (exploded view) of micro-fluidic chip mould among the present invention.
Fig. 3 makes the noumenal modification polymethyl methacrylate micro flow control chip synoptic diagram for the original position bulk polymerization.
Fig. 4 is the encapsulation of micro-fluidic chip.
Fig. 5 is three kinds of environmental contaminants 2 of methacrylic acid noumenal modification polymethyl methacrylate micro flow control chip separation detection of the present invention, the electrophoresis pattern of 6-xylenol (a), phenol (b) and parachlorophenol (c).
Number in the figure: 1 is the sample solution hole, 2 for separating microchannel, 3 for containing microchannel 2 and 7 and solution hole 1,4,5 and 6 micro-fluidic chip, 4,5 and 6 are the buffer solution hole, 7 is the sample introduction kapillary, 8 and 8 ' is glass plate, and 9 for middle hollow out is the aluminium sheet of the rectangle of chip size, and 10 is the chip size cavity, 11 is open slot, 12 is the silicon formpiston, 13 microchannels for silicon male mold surfaces protrusion, and 14 is the micro-fluidic chip mould, 15 are the surperficial micro-fluidic chip substrate that microchannel is arranged, 16 microchannels that fall in for the micro-fluidic chip substrate surface, 17 is epiphragma, 18 is micro-fluidic chip.
Embodiment
Further describe the present invention below by embodiment and accompanying drawing:
1, the making of methacrylic acid noumenal modification polymethyl methacrylate micro flow control chip
(A) capillary channel and the solution connection holes of Adobe Illustrator 10.0 software design chips adopted in the design of electrophoresis chip, adopt high resolving power (3600dpi) laser photocomposing system on the polyester transparent film, to be printed as the mask negative film, microchannel width on the mask is 50 μ m, solution connection holes is the circular hole of diameter 2mm, wherein microchannel (separation capillary 2 and sample introduction kapillary 7) and solution hole 1,4,5 and 6 (Fig. 1) are transparent, and remainder is a black.The erect image of micro-fluidic chip is seen Fig. 1.Separate microchannel 2 long 6.0cm, sample introduction microchannel 5 long 0.5cm, wherein kapillary 4 and 5 point of crossing are 0.5cm to the distance of three nearest solution connection holes.
(B) silicon chip (the p type of being produced on of silicon negative film and chip mould through oxidation processes, thick 500 μ m, 4 inches of diameters, crystal orientation＜100 〉, surface silica dioxide oxidation bed thickness 100nm) by spin-coating technology (rotating speed 3000rpm, 40 seconds) coating one deck negative photoresist (SU-8 photoresist), baking 40 minutes (preceding baking) in 65 ℃ of baking ovens, cover mask (the chip capillary microfluxion that contains design) then, (1.5-2mm) compresses with quartz glass plate, exposure 30min (365nm under ultraviolet ray, 45W), baking 25 minutes (baking of exposure back) in 65 ℃ of baking ovens, with the supporting developer immersion treatment of SU-8 after 90 seconds, rinsing 20 seconds is with the flush away photoresist layer of portion's exposed portion (zone beyond kapillary and the solution hole) not in acetone and isopropyl alcohol respectively, and baking 10 minutes (back baking) makes the photoresist sclerosis that capillary channel and solution connection holes partly expose on the silicon chip in 150 ℃ of baking ovens then, and silicon chip is dipped in 0.5M HF-0.5M NH
43.5 minutes SiO of F solution with silicon chip surface
2Layer etching off; then in 60 ℃ of about 1 hour of silicon chips (etching speed is 0.35 μ m/ minute) that expose with 40%KOH aqueous solution (containing 5% isopropyl alcohol) etching; form the outstanding capillary channel and the solution pore structure part of anode membrane; the silicon chip not photoresist on etched portions surface can come off in etching process automatically; silicon dioxide layer under the photoresist can not protected the silicon structure under the silicon dioxide layer by 40%KOH aqueous solution etching, promptly gets and makes the silicon chip formpiston.To have the silicon chip one side of kapillary tongue clamp one with one flat plate glass in the middle of hollow out be the aluminium sheet (about 2mm is thick) of the rectangle of chip size, the chip mould.
(C) injection molding and polymerization mix in methyl methacrylate monomer and a certain amount of dressing agent methacrylic acid (1-3% of the amount of monomer) glass container, add small amount of thermal initiating agent azoisobutyronitrile (methyl methacrylate monomer amount 0.15%) and a little light initiating agent styrax (methyl methacrylate monomer amount 0.15%) then, in 50 ℃ of water-baths heating and shake and make its dissolving, in 80-90 ℃ of water-bath, heated 15-20 minute then, shook mixed solution once in per 5 minutes, and made molten this pre-polymerization of monomer become glycerine shape clear solution.Attention will prevent that sealing enters in pre-collecting process, avoid temperature too high simultaneously, otherwise can be sudden and violent poly-because of causing, and causes the waste of material.Pre-polymerization later stage polymerization speed is accelerated, and polymerization should be controlled in 20 minutes in the time of 85 ℃, otherwise the polymeric solution viscosity of pre-polymerization is excessive, is difficult for pouring into mould and has easily introduced bubble.Above-mentioned pre-gathering solutions is injected mould 14 by mould openings groove 11, and open slot 11 caused bulk polymerization with 20W uviol lamp (365nm) in 30-60 minute by mould glass irradiation pre-gathering solutions up.Simultaneously respective liquid potpourri injection molding is about polymerization between the sheet glass of 100mm in the slit, obtains the epiphragma 17 of same material after the demoulding.
(D) after the demoulding and overlay film are treated the polymeric solution sclerosis of pre-polymerization, with mould demoulding in ultrasonic 10 minutes in 40 ℃ of water-baths.( solution connection holes 1,4,5 and 6 is seen Fig. 1 with the boring of micro-fluidic chip substrate 15 channel ends of the demoulding, aperture 2mm) is used to connect solution, for prevent that microchannel from stopping up in encapsulation process, before the overlay film, in the microchannel of substrate, fill low-melting water-soluble polymers polyglycol (PEG) 1500, fusing point is 43-49 ℃, because of the low easy fusing of fusing point can be filled in the microchannel under liquid state, behind the cooling curing, use wet filter paper wiping micro-fluidic chip substrate surface behind the cooling curing to remove the outer PEG of passage.With chloroform epiphragma 17 is bonded on the micro flow chip substrate 15 again, in the water-bath that is higher than the filled polymer fusing point, the microchannel interpolymer is melted then, by the hot water in the syringe it is released microchannel and also cleaned outward, promptly can be made into the noumenal modification polymethyl methacrylate micro flow control chip finished product.This method can reduce the rejection rate of micro-fluidic chip preparation.The one side that epiphragma is arranged is a chip upper surface.The length of capillary electrophoresis chip 1 is 6.5mm, and width is 2.0mm, and thickness is 2.2mm.
The chip of this method preparation successfully is used to detect three kinds of phenolic comp ' ds pollution (see figure 5)s
2, to the making of vinylpyridine noumenal modification polymethyl methacrylate micro flow control chip
The making of the design of electrophoresis chip and silicon formpiston and chip mould is with embodiment 1.Methyl methacrylate monomer is mixed in vinylpyridine (1-3% of the amount of monomer) glass container with a certain amount of dressing agent, add small amount of thermal initiating agent azoisobutyronitrile (methyl methacrylate monomer amount 0.15%) and a little light initiating agent styrax (methyl methacrylate monomer amount 0.15%) then, in 50 ℃ of water-baths heating and shake and make its dissolving, in 80-90 ℃ of water-bath, heated 15-20 minute then, shook mixed solution once in per 5 minutes, and made molten this pre-polymerization of monomer become glycerine shape clear solution.Attention will prevent that sealing enters in pre-collecting process, avoid temperature too high simultaneously.Pre-polymerization later stage polymerization speed is accelerated, and polymerization should be controlled in 20 minutes in the time of 85 ℃, otherwise the polymeric solution viscosity of pre-polymerization is excessive, is difficult for pouring into mould and has easily introduced bubble.Above-mentioned pre-gathering solutions is injected mould 14 by mould openings groove 11, and open slot 11 caused bulk polymerization with 20W uviol lamp (365nm) in 30-60 minute by mould glass irradiation pre-gathering solutions up.Simultaneously respective liquid potpourri injection molding is about polymerization between the sheet glass of 100mm in the slit, obtains the epiphragma 17 of same material after the demoulding.After treating the polymeric solution sclerosis of pre-polymerization, with mould demoulding in ultrasonic 10 minutes in 40 ℃ of water-baths.The micro-fluidic chip substrate 15 channel ends borings ( solution connection holes 1,4,5 and 6 is seen Fig. 1, aperture 2mm) of the demoulding are used to connect solution, and embodiment 1 is seen in the encapsulation of micro-fluidic chip.
It is in 5 the acetate buffer solution that experiment is presented at pH, to vinylpyridine noumenal modification polymethyl methacrylate micro flow control chip electroosmotic flow direction from the negative pole to the positive pole, opposite with the chip electroosmotic flow direction among the embodiment 1, show surperficial lotus positive electricity to the vinylpyridine noumenal modification polymethyl methacrylate micro flow control chip, this be because in the chain of polymethylmethacrylate, introduced lotus positive electricity to the vinylpyridine residue.Based on positive electric group is to the effect of negative ion in the passage, practicality shows that this chip can improve organic acid electrophoretic separation such as acetate, tartrate and oxalic acid.
Claims (3)
1, a kind of preparation method of noumenal modification polymethyl methacrylate micro flow control chip, close by functional dressing agent and methyl methacrylate in-situ copolymerization, dressing agent is introduced the main chain of polymethylmethacrylate, charge to the polymethylmethacrylate surface is regulated and control, it is characterized in that concrete steps are as follows: make the micro-fluidic chip mould earlier: adopt computer aided design software design micro-fluidic chip, make the silicon chip formpiston that contains the microchannel projective structure by photoetching and chemical corrosion technology, it is the aluminium sheet of the rectangle of chip size with two sheet glass clampings, one middle hollow out that the silicon chip one side of microchannel projective structure will be arranged, and constitutes the chip mould; Thermal initiator azoisobutyronitrile and light trigger styrax are dissolved in the mixed solution of monomers methyl methacrylate and dressing agent methacrylic acid or vinylpyridine, and heating is 15-20 minute in 80-90 ℃ of water-bath, makes the monomer solution pre-polymerization; Pre-gathering solutions is injected the said chip mould, use UV-irradiation pre-gathering solutions 30-60 minute, cause bulk polymerization, make the micro-fluidic chip substrate by the mould glass; Adopt bonding encapsulation technology again, epiphragma is bonded on the micro-fluidic chip substrate, promptly can be made into the micro-fluidic chip finished product.
2, preparation method according to claim 1 is characterized in that the dressing agent consumption is the 1-3% of monomer weight, and the consumption of thermal initiator and light trigger is respectively the 0.1-0.2% of monomer weight.
3, preparation method according to claim 1, it is characterized in that adopting the bonding encapsulation technology of water-soluble low melting point polymer secondary solvent, before the chip substrate overlay film, in the microchannel, fill wax shape polyethylene glycol 1500 by knife coating, in 45-55 ℃ of baking oven, make its fusing; The cooling back uses wet filter paper wiping micro-fluidic chip substrate surface to remove the outer polyglycol of passage; With chloroform epiphragma is bonded on the micro flow chip substrate, room temperature was placed after 12-15 hour, in 70-80 ℃ of water-bath, make the polyglycol fusing in the microchannel, by the hot water in the syringe it is released microchannel and also cleaned outward, promptly can be made into the micro-fluidic chip finished product.
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| CN101585508B (en) * | 2009-07-02 | 2011-12-07 | 复旦大学 | Preparation method of organic glass micro-fluidic chip based on photosensitive thixotrope film |
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| CN101424660B (en) * | 2007-10-31 | 2012-01-04 | 南京大学 | Design and production of portable highly effective capillary pipe electrophoresis chip sampling by chink |
| CN101654654B (en) * | 2009-09-16 | 2012-07-04 | 华东师范大学 | Method for modifying DNA microfluidic chip micro-channel with mixed screening medium |
| CN102452639A (en) * | 2010-11-01 | 2012-05-16 | 香港理工大学 | Binding method of plastic microfluidic chip and plastic microfluidic chip |
| CN102166537B (en) * | 2011-01-30 | 2013-05-01 | 南京大学 | Hydrophilic, multifunctional and integrated miniflow control chip easy to optical detection, manufacture method thereof and use thereof |
| CN103437240B (en) * | 2013-09-13 | 2015-10-28 | 中国科学院长春应用化学研究所 | Paper chip and preparation method thereof |
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| CN1464303A (en) * | 2002-06-20 | 2003-12-31 | 中国科学院理化技术研究所 | Process for preparing high polymer micro-flow control chips |
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| US5258503A (en) * | 1987-09-08 | 1993-11-02 | Kanegafuchi Kagaku Kogyo Kabushiki Kaisha | Autoantibody adsorbent and apparatus for removing autoantibodies using the same |
| JP2003098060A (en) * | 2001-09-26 | 2003-04-03 | Olympus Optical Co Ltd | Method for selectively collecting target area using probe microscope, method of making substance contained in target area react, and device therefor |
| CN1464303A (en) * | 2002-06-20 | 2003-12-31 | 中国科学院理化技术研究所 | Process for preparing high polymer micro-flow control chips |
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| CN101585508B (en) * | 2009-07-02 | 2011-12-07 | 复旦大学 | Preparation method of organic glass micro-fluidic chip based on photosensitive thixotrope film |
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