CN107335478A - Hard modifying interface is relied on to reduce the cheap micro fluidic device of cholera diagnosis of flow resistance - Google Patents

Abstract

The present invention relates to a kind of cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis, belong to analysis testing field.Serial problem be present with dimethyl silicone polymer i.e. PDMS that is cheap and easily processing come the substrate for making cholera diagnosis micro-fluidic chip；The PDMS has spontaneous slowly deformation tendency, and the spontaneous slowly deformation tendency can cause its pipeline of chip to become narrow gradually, and PDMS surfaces are strongly hydrophobic, and surface chemical modification modified effect is difficult to persistently, its surface and can also adsorb, swallow up large biological molecule.This case main points are that substrate material is the PDMS for having ecosystem surface, and micron silica particle is filled with its pipeline of chip；Its function of the micron silica granule filler includes supporting pipeline inwall, and, with the set of hydrophily fluid channel come the hydrophobic pipeline of compensatory script, and, by hard squeeze, cover the chance for thereby reducing large biological molecule and being contacted with the internal face in inner-walls of duct face.

Description

Hard modifying interface is relied on to reduce the cheap micro fluidic device of cholera diagnosis of flow resistance

Technical field

The present invention relates to a kind of cholera for relying on hard modifying interface to reduce flow resistance diagnosis cheap micro fluidic device, the micro fluidic device It is that analysis testing field is belonged to come the special purpose device of cholera diagnosis antigen based on antigen/antibody specific reaction.

Background technology

The micro-fluidic cholera diagnostic techniques background of associated multi-channel, it may refer to the grade applications for a patent for invention of CN 200910150930.4 Case.

Only for the microflow control technique overall general picture of itself, it may refer to famous micro-fluidic expert Mr. Lin Ping Cheng and go out not long ago Monograph " diagram Microfluid based Lab on a chip ", the monograph is published via Science Press, and the monograph is to microflow control technique Past, now, and, vision of the future etc. etc., suffer from detail, be deep into long discussion of detail.

So, the Important Problems of this case that to have a talk below concern.

The basic framework of micro-fluidic chip, including it is etched with the substrate of conduit and the cover plate to fit together therewith, the substrate On fluid course, before upper cover plate is assembled, it is apparent on see to be exactly some conduits, wait until cover cover plate thereon after, Just really closure forms the fluid course, and the conduit inner surface of the conduit is together with the part cover plate structure that surround the conduit Into described fluid course；So, it is clear that assemble the fluid course after completing, the main portion of its inner surface area Point it is the inner surface area of that conduit, in other words, the state or property of the conduit inner surface substantially determine that the liquid stream is led to The integrality or property in road；Therefore say, the inner surface state or inner surface property of this conduit of the structure on substrate are crucial Factor；In principle, any material that can be kept or keep its solid forms substantially, can be used to make substrate and lid Piece, such as, the material that can act as substrate and cover plate can be for example poly- diformazan of monocrystalline silicon piece, quartz plate, sheet glass, high polymer Radical siloxane, polymethyl methacrylate, makrolon etc.；Certainly, the selection of substrate and the selection of cover plate can with identical, It can also differ；From the point of view of material consumption, manufacture difficulty and application popularization prospect etc. etc., exist between these materials Not small difference, the especially selection of that substrate, have a great influence.

In various substrate making materials, dimethyl silicone polymer, i.e. PDMS, comparatively very easily shaping, so Substrate on to make conduit extremely simple, and the lower cost for material, substrate is made with the polydimethyl siloxane material, Conduit is suppressed or etched thereon, and is engaged with the cover plate that the cheap material such as glass or polypropylene or other plastic sheets makes, seemingly It is a kind of more satisfactory selection；Certainly, patch material can also select to use cheap polydimethyl siloxane material：So, This substrate selection is the scheme of polydimethyl siloxane material, and material is extremely cheap, and making is extremely simple, seems and should also be as being extremely easy to Popularization, promote.

But thing is really not so simple.

First, this polydimethyl siloxane material, that is, the material that alphabetical PDMS is referred to of abridging, itself are a kind of strong Hydrophobic material, conduit is built on this material, if without being operated for the modified of the conduit surface, then, it is overall After assembling is completed, that is, after covering cover plate, because the conduit its inner surface in structure is occupied in most fluid course Surface, then, its strong hydrophobic property of the PDMS conduits inner surface, is deciding factor, and it can be similar to water-soluble The fine liquid stream of polar liquid of liquid by becoming very difficult, its flow resistance is big, or even in general Micropump is all difficult to promote, Certainly, if cover plate also selects to use the PDMS material, then, problem is substantially identical, similar；Therefore, existing Among having technology, modification is modified particular for the conduit inner surface on the PDMS material, is necessary operation；So, This is pretty troublesome for the modified operation of PDMS conduit inner surfacesThat falls nor this problem, forms serious technical puzzlement , it is another problem：PDMS polymer molecules inside its body phase of this PDMS material substrate, which have from trend surface, to be expanded Dissipate, the characteristic of migration, the characteristic that PDMS polymer molecules are diffused to the surface, migrated automatically inside this substrate body phase, will make The sufficiently long time must can not be maintained by the state after the modification of its inner surface of that conduit of surface modifying and decorating, that Holding time for conduit its inner surface state after surface-modified is substantially only sufficient to the time for completing laboratory internal test experiments Need；In other words, by surface is modified or the PDMS conduit inner surfaces of surface modification, after it is modified or modification is said The surface state formed afterwards can not be lasting, but soon automatically tends to or say the surface state before becoming surface modification again, The strong hydrophobic surface state of that script is returned in the shorter time, then, just think, such micro-fluidic chip energy Enough a large amount of making, mass storages, it is widely popularized, answer is it is obvious that is, impossible.On this PDMS material Conduit, if not doing surface modification, it can not pump and pass through similar to the fine liquid stream of polar solvent of the aqueous solution, chip also just cannot Use；And if having done surface modification, its state after modifying can not be persistently kept again, or equally can not popularization and application.

According to the literature, there is a kind of expedient solution, be that a small amount of surfactant is added in sample to be tested solution, with Just during test, dynamic, temporary transient effective surface hydrophilic layer is built temporarily in the PDMS conduits inner surface；However, Due to the amphipathic characteristic of surfactant, the introduced surfactant of the program is inevitable simultaneously also can be with being tested in sample solution Combination occurs for composition so that tested composition can not be detected normally or it is detected ratio and dubiously reduced；Live on the surface Property agent is possibly even fully wrapped around by tested composition with micelle form so that tested composition can not be detected or identify；Therefore, should Kind adds the scheme of surfactant into sample solution, is far from a preferable solution.

So, how to accomplish that substrate can either be made using cheap PDMS material, and the conduit inner surface can be released Decorating state can not persistently, chip can not largely make, largely lay in and then be widely popularized and such a make the numerous specialties in this area The puzzlement that personnel are entangled with for a long time, the highly difficult problem that exactly one its obvious technology barrier can not despise.

Be present many year in the highly difficult problem, so far, not yet properly settled.

Second, the PDMS material of non-surface modification, it is stated that, its surface is strongly hydrophobic above, this strong hydrophobic Material surface and also another problem, that is, this strong hydrophobic PDMS surfaces can adsorb large biological molecule, and And these adsorbed large biological molecules can also further depression further on PDMS surfaces, gradually fall into it is gradually deep, directly It is trapped within the body phase of PDMS substrates to heavy, in fact, this process, is partly also due to inside PDMS material body phase Polymer molecule, which has, to be diffused to the surface, caused by travel motion；Such case, it can also be explained from another angle, i.e. Continuously from inside PDMS body phases to those polymer molecules of its diffusion into the surface, migration, its result moved, be by Gradually those are involved within the body phase of PDMS substrates by the large biological molecule of adsorption, briefly, these are inhaled Attached large biological molecule is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up large biological molecule Phenomenon, the influence caused by it, necessarily cause the severe deviations of all kinds of test data of experiment for being related to large biological molecule.

As described above, the problem of PDMS substrates, is, its not only adsorption large biological molecule, and swallow up large biological molecule, So, as the large biological molecule of experiment test object, its disappearance will not stop because surface saturation is adsorbed, but, no It is disconnected adsorbed, also constantly swallowed up.

On PDMS substrates in related experiment test process its body phase constantly swallow up test associated biomolecule macromolecular phenomenon, separately It is to say that one kind, which is explained, substantial amounts of Minute pores in PDMS body phases be present, and associated biomolecule macromolecular is by after adsorption, depression Into these Minute pores, and then swallowed up；However, inventor thinks, those can allow the air point of miniature scale Son squeezes into the Minute pores therebetween, not equal to saying that they also can directly allow the large biological molecule of relative large scale to enter, two Person's difference on yardstick is huge, must not make sweeping generalizations.Explanation is bypassed, in any case, the biology as dependence test analysis object Macromolecular is adsorbed by PDMS substrate conduits inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is known objective deposits Phenomenon.

, can be from containment PDMS surfaces to life in order to prevent swallow up effect of this PDMS substrate bodies relative to large biological molecule The absorption of thing macromolecular addresses, and method is chemically modified modification aiming at the PDMS material surface, for For PDMS is the situation of substrate material, modification exactly is chemically modified to the surface of described channel portion, by chemistry The conduit inner surface of modification, can contain its absorption to large biological molecule, and then avoid large biological molecule quilt PDMS substrate body phases are swallowed up；But or that old problem, that is, the chemical modification on PDMS material surface changes Surface state after property can not persistently be kept, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface automatically, The process of migration, soon it can become that conduit inner surface state being modified by surface chemical modification again script strong hydrophobic And the state of strong adsorption large biological molecule, in other words, no matter how professionals in the field turn from side to side, the PDMS bases Its conduit inner surface of piece is always rapidly to strong hydrophobic surface state evolution.

So, how can either obtain that PDMS material price is extremely cheap, substrate makes extremely easy benefit, and can enough reaches Growth stage contains the absorption process of the PDMS substrate conduit inner surfaces to large biological molecule, and then prevents PDMS substrate bodies relative The effect of swallowing up of large biological molecule so that related chip manufactured goods are able to maintain that a prolonged enough, rational shelf-life, It is exactly a very intractable problem.The problem equally makes the numerous specialties in this area as another problem addressed above Personnel are entangled with, perplexed for a long time, and the problem is equally the highly difficult problem that its obvious technology barrier can not despise.The hardly possible Also be present many year in topic, so far, also not yet properly settled.

Third, the polydimethyl siloxane material, namely PDMS material, the active force very little wherein between polymer molecule, its The polymer molecule of low polymerization degree inside body phase is in constantly into the dynamic process of surface migration, therefore, includes PDMS bases After it has been formed, the surface topography in its inner-walls of duct face of chip can be because of described low polymerization degree polymer molecule not for the chip of sheet material Migrate disconnectedly, pour into the surface and change, the change of its pattern is somewhat similarly to Rheological Deformation, trickling deformation or deformation of creep institute Pattern changes caused by possible；In the higher PDMS material of some low polymerization degree component ratios, this kind is somewhat similarly to flow The phenomenon of deformation, trickling deformation or the deformation of creep can show must be than more significant；Its shaping of chip of this kind comprising PDMS substrates The dynamic of inner-walls of duct surface topography afterwards changes property, namely the described property for being somewhat similarly to Rheological Deformation or trickling deformation Property or the deformation of creep property, this case applies mechanically the word of rheology one, integration, also referred to as the rheological equationm of state or Rheological Deformation property, The dynamic of corresponding inner-walls of duct face pattern changes, and this case is also referred to as rheology；As described above, because this kind of material its own The certain Rheological Deformation property as described above having, its manufactured goods actually can at leisure Rheological Deformation and cause the manufactured goods whole Individual structural form changes；Influence caused by its Rheological Deformation property, in terms of macroscopic perspective be it is subtle, still, For the very fine micro-fluidic chip of its internal structure, the rheological equationm of state can but cause bigger influence, therefore, in advance The fluid course namely the pipeline formed when first making, because the wall of its tube chamber of pipeline is mainly PDMS materials Wall, so, the cross-sectional area of the pipeline can actually taper into because of rheology over time, in other words, Due to the rheology, can more it become more for natively finer its tube chamber of the pipeline that liquid sample flows through Narrow, in the case of flow time is sufficiently long, or even the tube chamber of the pipeline can partly be closed or fully closed, and then be caused Sample fluid course is thoroughly blocked, the journey for closing or fully closing with being even also not reaching to its luminal part of pipeline Degree, its fluid passage that can be provided of that pipeline narrow because of rheology, then, in pipeline, its inner surface is exactly originally strong It is strong it is hydrophobic in the case of, aqueous sample liquid is just more difficult to by becoming narrower micro-channel so because of rheology, then, The influence of substrate rheology how is resisted, maintains the basic framework of the pipeline that gross distortion does not occur for a long time, also, keep the pipeline Long-term unobstructed, the problem of being exactly a urgent need to resolve.

The content of the invention

The technical problem to be solved by the invention is to provide a package solution, solve what is addressed above totally The problem of three aspects, also, the solution is applied to cholera diagnosis multichannel micro-fluidic chip field, formed a kind of Multichannel micro-fluidic device is used in new cholera diagnosis.

The present invention solves the technical problem by following scheme, and the device that the program provides is a kind of hard modifying interface drop of support The cholera of low flow resistance diagnoses includes multichannel micro-fluidic chip, the miniflow with cheap micro fluidic device, the structure of the micro fluidic device The structure of control chip includes being bonded to each other the substrate and cover plate of installing together, and the substrate and cover plate are plate object or sheet The channel structure via mould pressing process or etching technics formation is contained in thing, that face towards the cover plate of the substrate, and the substrate is also Containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or simple drilling technology formed window Mouth structure, it is bonded to each other the substrate that is installed together and the cover plate and has been built into containing pipeline configuration and has been attached thereto jointly The micro-fluidic chip of liquid pool structure, the locations of structures of the pipeline are located at the interface zone that the substrate is bonded to each other with the cover plate, the window Its side of mouth is blocked by the cover plate and opposite side opens, and the locations of structures of the window is exactly the locations of structures of the liquid pool, the liquid The quantity in pond is three, and its locations of structures of two liquid pools therein is located at the sample introduction end of the micro-fluidic chip, a remaining liquid pool Its locations of structures is located at the terminal of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal and the sample introduction end Be located remotely from each other, one end of the pipeline via be similarly positioned in the manifold shape turnout of the interface zone being bonded to each other respectively with positioned at sample introduction The Liang Ge liquid pools UNICOM at end, the other end of the pipeline and a remaining liquid pool for the terminal positioned at the micro-fluidic chip UNICOM, and, the working electrode that is sequentially respectively installed in the pipeline on diverse location and to electrode and reference electrode, institute It is sensitive by conductive electrode and the gold size for having embedded cholera specific antibody being attached on the conductive electrode to state working electrode Film is formed, and the construction of the pipeline is in parallel construction, and the pipeline in parallel construction is made up of four lateral parallel connections, described The quantity of working electrode is four, and the installation position of four working electrodes is located in four laterals respectively, and, Specific antibody in its top layer gold size sensitivity membrane structure of four working electrodes can be specifically bound to cholera antigen respectively Four kinds of cholera antibody materials, four kinds of antibody materials are cholera TP0821 antibody and cholera TP0319 antibody and suddenly respectively Random TP0624 antibody and cholera O139 mycoprotein antibody, its material of the working electrode are that gold material or thermal decomposition are led Sheet or thread is presented in electric macromolecule material, the working electrode its pattern, and, Micropump, the Micropump and the micro-fluidic chip Sample introduction end connection, the Micropump is the Micropump of external form or the Micropump of built in version, and emphasis is that its material of the substrate is poly- Dimethyl siloxane material, its surface of the substrate are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not have By any surface chemical modification or the surface of the primary form of the material of any surface chemical modification；And many dioxies Silicon carbide particle, many silica dioxide granules are filled in the pipeline, and its tube chamber of the pipeline is by many silica Particle is filled up, and between 10 microns and 200 microns, the silica dioxide granule is its particle size range of the silica dioxide granule Its shared percentage by weight of silica crystalline particles, silica glass granule or silica composition is inorganic more than 80% Glass particle.

Its particle diameter of the silica dioxide granule can be that the basis between 10 microns and 200 microns is actually needed and is arbitrarily designated Particle diameter, such as 10 microns, 30 microns, 70 microns, 100 microns, 150 microns or 200 microns of the particle diameter, etc..

The scope of further preferred its particle diameter of the silica dioxide granule is between 100 microns and 200 microns.

Its pattern of the silica dioxide granule is unlimited, and the silica dioxide granule for example can be spheric granules, rod-shaped particles, rectangular Shaped particles or any amorphous granular.

The various silica dioxide granules manufacturing technology of itself is known technology.

Various forms, the silica dioxide granule market of all size are on sale.

The silica dioxide granule can also customize to specialized factory.

The preferred scope of its internal diameter of pipeline is between 500 microns and 1000 microns；It is but excellent compared to above-mentioned The internal diameter of the pipeline scope of choosing, more tiny internal diameter of the pipeline or thicker internal diameter of the pipeline are also what this case was allowed.

The word of Micropump one art-recognized meanings of itself are known for the professional in micro-fluidic chip field.

The Micropump both can be the Micropump of external form；The Micropump can also be done into or say inside the embedded micro-fluidic chip The Micropump of the built in version of its sample introduction end structure position or the sample introduction end Near-neighbor Structure position, its basic structure key element and external form Micropump it is identical.

For example miniature piezoelectric pump of the Micropump, miniature peristaltic pump or miniature air driven pump, etc..

Involved Micropump can customize to specialized factory.

Its connected mode with various micro-fluidic chip of various Micropump is known.

Fluid course described in pipeline described in this case and this case, this two kinds to express its art-recognized meanings identical.

The gold size sensitive membrane is to be sufficiently mixed chitosan gold size solution and cholera specific antibody solution uniformly, uses point sample instrument Point sample is coated on specified structure position, and forms its drying and forming-film.Cholera specific antibody in the gold size sensitive membrane is equal The cholera antibody marked for horseradish peroxidase or glucose oxidase, the gold size sensitive membrane have been included as fixing above-mentioned each Cholera specific antibody and introduce complementary medium therein, the complementary medium such as chitosan, cellulose acetate, gelatin Therein a kind of or their mixture.

The pipeline and the lateral and the manifold shape turnout in the microfluidic chip structure, its internal diameter chi It is very little to may each be arbitrarily selected size, still, for prepare liquid sample and reducing the consideration of reagent loss etc. less as far as possible, The passage of capillary level, the capillary level are preferably selected in the pipeline and the lateral and the manifold shape turnout Passage imply that its internal diameter passage suitable with the internal diameter of the capillary on ordinary meaning.Its inner passage of capillary it is transversal Face shape can be arbitrary shape, the shape of cross section is for example circular, oval, square, rectangle, bar shaped, certainly Can the linear of bending arbitrarily be present, also, the interior shape of the capillary is with the extension of pipeline, different parts Shape of cross section can also allow to be different shapes.Only for the word of capillary one, its art-recognized meanings is known.

What is be related in structure is the electrode of microsize to electrode and reference electrode, and its electrode shape may each be any choosing Fixed shape, the arbitrarily selected shape such as square piece shape, rectangular patch, strip or round sheet etc..It is described to electricity The art-recognized meanings of pole and the reference electrode vocabulary of itself are known.

It is related to several liquid pools in this case microfluidic chip structure, the liquid pool is the pond shape or scrotiform structure for transitional liquid storage Make, its shape of the inner chamber of each liquid pool may each be arbitrarily selected shape, the cavity shape such as cylindrical empty Cavity-like, square column type cavity-like, oblong cavity shape or spherical hollow space shape etc..The size of the liquid pool can be arbitrarily selected Size, still, in order to which less with prepare liquid sample and reduction reagent loss, the liquid pool is preferably capable of and capillary as far as possible The liquid pool of the microminiature of matching.

Only with regard to naked PDMS substrates itself conduit molding or lithographic technique for, be open-and-shut known technology；Equally Ground, the technology of hole-opening is even more known simple technique on naked PDMS substrates.This case is not also to this expansion superfluous words.

This case device can further include some annexes, the annex such as multiple tracks electrochemical workstation etc., institute certainly The art-recognized meanings for stating multiple tracks electrochemical workstation are known.Each working electrode for being related in this case microfluidic chip structure and , can be respectively via corresponding special the corresponding interface got lines crossed with the multiple tracks electrochemical workstation to electrode and reference electrode etc. Coupled.It is described that special to get lines crossed be for each the corresponding interface of each electrode and the multiple tracks electrochemical workstation is carried out into phase The private cable being mutually coupled with.The micro-fluidic chip in this case device, its structure can also include micro-valve, the number of the micro-valve Measure unlimited, according to being actually needed, the micro-valve can be installed in the position of any required installation in the microfluidic chip structure； For the word of micro-valve one for the professional of micro fluidic chip technical field, the art-recognized meanings of itself are known；The micro-valve The manufacturing technology of itself and the use of technology is also known；The component that the micro-valve is not required.

The diameter of the working electrode can allow to be that any setting is easily installed the suitable diameter used, it is however recommended to Or say preferable its scope of the diameter between 0.1 micron to 2000 microns；The length of the working electrode can permit Permitted to be that any setting is easily installed the length used, it is however recommended to or to say the preferable length its scope be at 1 micron To between 15000 microns.

The gold size that the working electrode surface layer is installed in by spraying or point sample instrument point sample or the coating of other appropriate process is quick Feel film, its thicknesses of layers can allow be any setting treat sample measuring liquid occur electrical signals response thickness, still, push away Preferable thickness is between 10 nanometers and 200 nanometers to the thickness recommended in other words.

The cover plate in chip structure, its material can allow to be any electrical insulating property material, such as：Polypropylene, glass Glass, polymethyl methacrylate, dimethyl silicone polymer, etc..

Positioned at the sample introduction end of the micro-fluidic chip two liquid pools its with positioned at the micro-fluidic chip terminal this is remaining Air line distance between one liquid pool can be arbitrarily selected suitable distance as needed, and still, the preferred value of the distance is Between 3 centimetres and 7 centimetres.

Described its thickness of cover plate and substrate can allow be any setting be easy to assembling thickness, the thickness of recommendation or say preferably Thickness be between 1.0 millimeters and 5.0 millimeters.Less thickness is advantageous to save material.

The application method of this case micro-fluidic chip：

This case is flowed with Micropump driving liquid stream in the capillary channel of the four-way micro-fluidic chip, utilizes four-way electrification credit Analyzer device is detected to four kinds of cholera diagnostic antigens respectively.

The specific detection of this case micro-fluidic chip is as follows using step：

1st, blood serum sample liquid is added in micro-pipe road, under Micropump driving, various cholera antigen molecules are by electric in each passage The cholera specific antibody capture for the corresponding horseradish peroxidase-labeled that gold size sensitive membrane embeds on the surface of pole.

2nd, the cholera specific antibody of horseradish peroxidase-labeled forms immune complex with the cholera antigen in blood serum sample.

3rd, using multi-channel electrochemical analyzer, the electron mediators such as catechol are added, using the above-mentioned reaction of amperometric detection Caused curent change, it is derived from the species and content of various analytes.

4th, result is subjected to comprehensive analysis, comprehensive diagnos is carried out to cholera antigen.

It is an advantage of the invention that its scheme of this case is will be all vacant in addition to electrode installation takes up space in the pipeline Space is all filled up with the silica dioxide granule.Those are filled in the silica dioxide granule in pipeline, and this case is overall referred to as to it Silica dioxide granule filler, its function include support, withstand the tube chamber of the pipeline its inwall so that the tube chamber of the pipeline is exempted from Narrow in because of the rheology, prevent its reduced cross-sectional area of the tube chamber of the pipeline or even close, so as to maintain the pipeline for a long time Basic framework serious deformation does not occur.

Numerous silica dioxide granule entities are mutually banked up under this case framework together at random in the pipeline, the silica The function of grain filler, also includes with the hydrophilic surface of its silica dioxide granule, utilizes each adjacent silicon dioxide particle certainly Mutual formed complications are continuous and its inner surface that is mutually close together by water-wetted surface has the sky of hydrophilic nmature Gap, in the hydrophilic fluid channel of the pipeline Inner Constitution network morphology, the having all the time although this complications can be continuous The fluid channel of hydrophilic nmature, then, inside the tube chamber of the pipeline simultaneously and a plurality of fluid channel deposited its synthesis, The effect of integrated, cumulative superposition in other words, is the equal of the relatively fine hydrophilic capillary channel of caliber；The complications Its presence of continuous hydrophilic fluid channel, counteracts its surface of the PDMS substrates of the primary form to a large extent The flow resistance for liquid sample caused by strong hydrophobic property；In other words, because of liquid sample its account for maximum ratio into It is water to divide, and liquid sample is substantially exactly the aqueous solution, and therefore, the presence of the silica dioxide granule filler can be significantly Overcome with the unmodified PDMS substrates of hydrophobic property and surface its to the aqueous solution it is incompatible, repel and barrier effect, And then be greatly reduced belong to properties of Aqueous Solution sample liquid stream its be advanced through the resistance of pipeline.

Under this case framework, inside the tube chamber of the pipeline, the area of its lumen wall wall of former pipeline, because of the titanium dioxide The rigid of silicon grain filler ties up, on lumen wall wall still can also be exposed its face of the surface of part hydrophobic property Product is already dramatically reduced, and the area on its still exposed part surface for belonging to hydrophobic property does not have far smaller than The wall surface area of the hydrophobic property of the lumen wall wall in the presence of silica dioxide granule filler；So, at this Under the case framework, in its lumen of the pipeline, its remaining hydrophobic surface increases surface newly with hydrophilic part, that it is superimposed, comprehensive , summation, cumulative technique effect, be to form the surface for tending to be hydrophilic on the whole；In other words, in the pipeline Inside tube chamber, its interfacial tension between sample solution of the part surface of remaining hydrophobic property, plus that of hydrophilic nmature Newly-increased its interfacial tension between sample solution of surface in part, that it is superimposed, comprehensive, summation, cumulative technique effect, Be so that its solid-liquid interfacial tension inside the tube chamber of the pipeline synergy level off to its tube chamber inner surface of capillary glass tube with Interfacial tension between sample solution.

Therefore, exist in the pipeline and stated under this case framework of silica dioxide granule filler, this case scheme just can be with Relatively low Micropump pump power and relatively low Micropump pumping pressure reach the driving to sample liquid stream.

On the other hand, due to its rheological equationms of state of PDMS as substrate material, the institute being filled in the pipeline State silica dioxide granule filler, can be gradually by constantly Rheological Deformation and the PDMS materials that are promoted to arest neighbors free space Pipeline wall more tightly wrap up, this process eventually causes those part silica for pressing close to the lumen wall wall Particle is stuck in original place or embedded in original place, the part be stuck in original place or embedding in-situ silica dioxide granule is mutual with remaining The silica dioxide granule banked up with being closely packed together each other incarceration together, due to this reason, the silica Grain filler just will not move in the pipeline easily, and silica dioxide granule therein, which is also substantially all, is locked in original place , therefore the stream framework of the micro-fluidic chip can be kept for a long time.

The particle size range of its particle of this case silica dioxide granule filler is between 10 microns and 200 microns, compared to general For organic molecule, general large biological molecule, the particle diameter of its particle of this case silica dioxide granule filler can be rated as it is huge incomparable, Due to its huge particle diameter, and the inner surface of the surface of its polarity and the pipeline of strong hydrophobic PDMS materials can not phase Perhaps say and mutually melt, then, the huge silica dioxide granule of this kind of particle diameter can not be by the pipeline of PDMS materials Surface is adsorbed, and can not more be swallowed up by the inner surface of the pipeline of PDMS materials, the huge dioxy of this kind of particle diameter Silicon carbide particle surely not be fully sunk in easily or sink to completely PDMS materials its among intrinsic many Minute pores, And because above it is stated that the rheology the reason for, the silica dioxide granule filler can be stuck in original place in other words Embedded in original place without moving easily.

Inside the tube chamber of the pipeline, its lumen wall wall of former pipeline is entirely the surface of hydrophobic property originally, as described above, Under this case framework, rigid because of the silica dioxide granule filler ties up, still can also be exposed on lumen wall wall Its area of the surface of part hydrophobic property be already dramatically reduced, its still exposed part for belonging to hydrophobic property The area on surface be far smaller than no silica dioxide granule filler in the presence of the lumen wall wall dredge The wall surface area of aqueous nature；The reason for because of the rheology, it is close to those silica dioxide granules of lumen wall wall, gradually Ground is partially embedded into or is partly absorbed among the lumen wall wall, its direction pipe of the silica dioxide granule among the state The part surface of cavity wall wall is with having occurred and that the lumen wall wall of matching deformation is brought into close contact, then, in tube chamber The part wall that the matching is deformed and is brought into close contact with silica particles of having occurred and that in wall wall is actually It can not be contacted with flowing through the sample solution of pipeline, that is to say, that the wall of this part can not be to flowing through the examination of pipeline Sample solution large biological molecule therein and organic molecule produce suction-operated.Based on above-mentioned mechanism, then because filling up described Its presence of the silica dioxide granule filler of pipeline, the overwhelming majority of its lumen wall wall of the pipeline are tightly hidden Lid, these described walls tightly covered can not contact with flowing through the sample solution of the pipeline, be only remainder The hydrophobic surface for not being brought into close contact by its surface of silica dioxide granule, tightly covering of fraction may be connect with sample solution Touch, in other words, under this case framework, large biological molecule and organic molecule in sample solution its with hydrophobic PDMS walls Direct touch opportunity significantly reduce, thus, the lumen wall faces of PDMS materials its to large biological molecule and organic molecule Adsorptive hindrance and swallow up interference be greatly reduced；As described above, this case scheme be advantageous to exclude the adsorptive hindrance and Swallow up interference, be advantageous to be lifted the reliability of correlation analysis test data.

As described above, this case silica dioxide granule filler, its function actually includes being similar to the pipe for PDMS materials Road inner surface carries out the effect of chemical modification, and functioning as its this respect is the inner surface of pipeline by the PDMS materials by dredging The surface modification of aqueous nature is into the surface of hydrophilic nmature, and still, its is hydrophilic with general, usual PDMS surface chemical modifications The chemical modification layer retention time be short, hydrophilic effect can not keep situation prolonged enough different, particle described in this case it is huge two Silicon oxide particle filler, its particle being stuck by rheology can not both be moved easily, and the particle of its huge particle diameter more can not Swallowed up easily by the inner-walls of duct of PDMS materials, therefore, its from this respect is similar to the technique effect of surface chemical modification On see, its technique effect of this case, be form it is permanent, can not erase, can not consume, not be corroded, not swallowed up, The hydrophilic surface reforming layer being not dissolved, it is just comparable to a kind of build in PDMS base materials its relevant surfaces in effect Permanent hydrophilic modifying superficial layer.

The technical scheme of this case dissolved totally address above to dimethyl silicone polymer substrate its apply related system Row technical barrier.Based on this case scheme, the very cheap polydimethyl siloxane material of this kind is just possible in micro-fluidic chip system Standby, production, using etc. field play bigger effect.

Brief description of the drawings

Fig. 1 is the schematic diagram of this case micro fluidic device embodiment chip part construction, it is shown that this structure is bowed The structural form of chip internal pipeline and each electrode and each related liquid pool under angle, do not depict in the figure Micropump with And other annexes.

In figure, 1,2,10 be the different liquid pool of three installation positions respectively, and 3 be manifold shape turnout, 4,7,11,14 It is that installation position is different but four laterals parallel with one another for forming UNICOM's structure in parallel respectively, 5 be to be installed in lateral 4 Specific antibody material in its interior top layer gold size sensitivity membrane structure is the working electrode of cholera TP0821 antibody, and 6 be installing The specific antibody material in its top layer gold size sensitivity membrane structure in lateral 7 is electric for the work of cholera TP0319 antibody Pole, 12 be that specific antibody material in its top layer gold size sensitivity membrane structure being installed in lateral 11 is cholera The working electrode of TP0624 antibody, 13 be the specificity in its top layer gold size sensitivity membrane structure being installed in lateral 14 Antibody materials are the working electrode of cholera O139 mycoprotein antibody, and 8 be to electrode, and 9 be reference electrode.

Embodiment

In this case that Fig. 1 is shown embodiment, the structure of the micro fluidic device includes multichannel micro-fluidic chip, and this is micro- The structure of fluidic chip includes being bonded to each other the substrate and cover plate of installing together, and the substrate and cover plate are plate object or sheet The channel structure via mould pressing process or etching technics formation is contained in thing, that face towards the cover plate of the substrate, and the substrate is also Containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or simple drilling technology formed window Mouth structure, it is bonded to each other the substrate that is installed together and the cover plate and has been built into containing pipeline configuration and has been attached thereto jointly The micro-fluidic chip of liquid pool structure, the liquid pool are liquid pool 1, liquid pool 2, liquid pool 10, and the locations of structures of the pipeline is located at the base The interface zone that piece is bonded to each other with the cover plate, its side of the window is blocked by the cover plate and opposite side opens, the structure of the window Position is exactly the liquid pool 1, liquid pool 2, the locations of structures of liquid pool 10, and the quantity of the liquid pool is three, two liquid therein Pond is the sample introduction end that liquid pool 1 and liquid pool 2 its locations of structures are located at the micro-fluidic chip, a remaining liquid pool be liquid pool 10 its Locations of structures is located at the terminal of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal and the sample introduction end phase It is mutually remote, one end of the pipeline via be similarly positioned in the manifold shape turnout 3 of the interface zone being bonded to each other respectively with positioned at sample introduction The liquid pool 1 at end and the UNICOM of liquid pool 2, this of the terminal of the other end of the pipeline with being located at the micro-fluidic chip are remaining One liquid pool is the UNICOM of liquid pool 10, and, the working electrode that is sequentially respectively installed in the pipeline on diverse location and to electricity Pole 8 and reference electrode 9, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded cholera The gold size sensitive membrane of specific antibody is formed, and the construction of the pipeline is in parallel construction, and the pipeline in parallel construction is by four points Branch pipe(tube) is in parallel to be formed, and the quantity of the working electrode is four, and the installation position of four working electrodes is located at described four respectively In bar lateral, four working electrodes are working electrode 5, working electrode 6, working electrode 12, working electrode 13 respectively, And the specific antibody in four working electrodes its top layer gold size sensitivity membrane structures be respectively can specificity knot to cholera antigen The four kinds of cholera antibody materials closed, four kinds of antibody materials be cholera TP0821 antibody and cholera TP0319 antibody respectively with And cholera TP0624 antibody and cholera O139 mycoprotein antibody, for specifically deploying, 5 be to be installed in lateral The specific antibody material in its top layer gold size sensitivity membrane structure in 4 is the working electrode of cholera TP0821 antibody, and 6 be dress The specific antibody material being located in its top layer gold size sensitivity membrane structure in lateral 7 is the work of cholera TP0319 antibody Electrode, 12 be that specific antibody material in its top layer gold size sensitivity membrane structure being installed in lateral 11 is cholera The working electrode of TP0624 antibody, 13 be the specificity in its top layer gold size sensitivity membrane structure being installed in lateral 14 Antibody materials be cholera O139 mycoprotein antibody working electrode, the working electrode 5, working electrode 6, working electrode 12nd, its material of working electrode 13 be gold material or thermal decomposition conducting polymer material, the working electrode 5, working electrode 6, Sheet or thread is presented in working electrode 12, working electrode 13 its pattern, and, Micropump, the Micropump and the micro-fluidic chip Sample introduction end connects, and the Micropump is the Micropump of external form or the Micropump of built in version, and emphasis is that its material of the substrate is poly- two Methylsiloxane material, its surface of the substrate are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not pass through Cross the surface of the primary form of the material of any surface chemical modification or any surface chemical modification；And many titanium dioxides Silicon grain, many silica dioxide granules are filled in the pipeline, and its tube chamber of the pipeline is by many silica Grain is filled up, and for its particle size range of the silica dioxide granule between 10 microns and 200 microns, the silica dioxide granule is two Inorganic glass of its shared percentage by weight of silica crystalline particles, silica glass granule or silica composition more than 80% Glass particle.

Micropump is depicted without in Fig. 1.

Also the associate members such as multiple tracks electrochemical workstation are depicted without in legend.

Each working electrode in this example structure and electrode and reference electrode via each special cable or can be said respectively Get lines crossed respectively with the corresponding cable interface of the multiple tracks electrochemical workstation as annex or saying interface connection of getting lines crossed.

The word of Micropump one art-recognized meanings of itself are known for the professional in micro-fluidic chip field.

The Micropump both can be the Micropump of external form；The Micropump can also be done into or say inside the embedded micro-fluidic chip The Micropump of the built in version of its sample introduction end structure position or the sample introduction end Near-neighbor Structure position, its basic structure key element and external form Micropump it is identical.

For example miniature piezoelectric pump of the Micropump, miniature peristaltic pump or miniature air driven pump, etc..

Involved Micropump can customize to specialized factory.

Its connected mode with various micro-fluidic chip of various Micropump is known.

Claims (9)

1. relying on hard modifying interface to reduce the cheap micro fluidic device of cholera diagnosis of flow resistance, the structure of the micro fluidic device is including more Channel microfluidic chip, the structure of the micro-fluidic chip include being bonded to each other the substrate and cover plate of installing together, the substrate and Cover plate is plate object or tablet, and that face towards the cover plate of the substrate is contained to be formed via mould pressing process or etching technics Channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or The window structure that simple drilling technology is formed, it is bonded to each other the substrate being installed together and has been built into jointly containing pipe with the cover plate The micro-fluidic chip of road structure and the liquid pool structure being attached thereto, the locations of structures of the pipeline is located at the substrate and the cover plate is mutual The interface zone of fitting, its side of the window is blocked by the cover plate and opposite side opens, and the locations of structures of the window is exactly the liquid The locations of structures in pond, the quantity of the liquid pool is three, and its locations of structures of two liquid pools therein is located at entering for the micro-fluidic chip Sample end, its locations of structures of a remaining liquid pool are located at the end of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested End, the terminal are located remotely from each other with the sample introduction end, and one end of the pipeline is via the manifold for being similarly positioned in the interface zone being bonded to each other Shape turnout respectively with the Liang Ge liquid pools UNICOM positioned at sample introduction end, the other end of the pipeline with positioned at the micro-fluidic chip the end A remaining liquid pool UNICOM at end, and, the working electrode that is sequentially respectively installed in the pipeline on diverse location and right Electrode and reference electrode, the working electrode by conductive electrode and be attached on the conductive electrode embedded cholera spy The gold size sensitive membrane of heterogenetic antibody is formed, and the construction of the pipeline is in parallel construction, and the pipeline in parallel construction is by four branches Pipeline is in parallel to be formed, and the quantity of the working electrode is four, and the installation position of four working electrodes is located at described four respectively In lateral, and, the specific antibody in its top layer gold size sensitivity membrane structure of four working electrodes is that cholera is resisted respectively Four kinds of cholera antibody materials of proper energy specific binding, four kinds of antibody materials are cholera TP0821 antibody and cholera respectively TP0319 antibody and cholera TP0624 antibody and cholera O139 mycoprotein antibody, its material of the working electrode are Sheet or thread is presented in gold material or thermal decomposition conducting polymer material, its pattern of the working electrode, and, Micropump should Micropump is connected with the sample introduction end of the micro-fluidic chip, and the Micropump is the Micropump of external form or the Micropump of built in version, its feature It is, its material of the substrate is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, the primary form Surface its be intended to refer to the primary form of the not material by any surface chemical modification or any surface chemical modification Surface；And many silica dioxide granules, many silica dioxide granules are filled in the pipeline, the pipeline Its tube chamber is filled up by many silica dioxide granules, and its particle size range of the silica dioxide granule is micro- with 200 between 10 microns Between rice, the silica dioxide granule is silica crystalline particles, silica glass granule or silica composition its shared weight Measure inorganic glass particles of the percentage more than 80%.

2. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is that the pipeline and the lateral and the manifold shape turnout are capillary channel.

3. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is that the thermal decomposition conducting polymer is the electric conductivity material formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated Material.

4. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the working electrode Length between 1 micron to 15000 microns.

5. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is that the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is, the cover plate its material in structure is dimethyl silicone polymer material.

7. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is, positioned at the sample introduction end of the micro-fluidic chip two liquid pools its with the remainder positioned at the micro-fluidic chip terminal A liquid pool between air line distance between 3 centimetres and 7 centimetres；Described its thickness of cover plate and substrate in structure is situated between Between 1.0 millimeters and 5.0 millimeters.

8. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is that its particle size range of the silica dioxide granule is between 100 microns and 200 microns.

9. the cheap micro fluidic device of the cholera for relying on hard modifying interface to reduce flow resistance diagnosis according to claim 1, it is special Sign is that its inside diameter ranges of the pipeline are between 500 microns and 1000 microns.