Background technology
The micro-fluidic AIDS diagnosis technical background of associated multi-channel, it is special to may refer to the grade inventions of CN 200910151219.0Sharp application case.
Only for the microflow control technique overall general picture of itself, famous micro-fluidic expert Mr. Lin Ping Cheng may refer to notThe monograph " diagram Microfluid based Lab on a chip " gone out before long, the monograph is published via Science Press, and the monograph is for miniflowPast of control technology, now, and, vision of the future etc. etc., suffer from detail, be deep into long opinion of detailState.
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, instituteState the fluid course on substrate, before upper cover plate is assembled, it is apparent on see to be exactly some conduits, wait until lid is covered thereonAfter piece, just really closure forms the fluid course, and the conduit inner surface of the conduit is together with the part that surround the conduitCover plate forms described fluid course together；So, it is clear that assemble the fluid course after completing, its inner surface faceLong-pending major part is the inner surface area of that conduit, and in other words, the state or property of the conduit inner surface are substantially determinedThe integrality or property of the fluid course are determined；Therefore say, the inner surface state or interior of this conduit of the structure on substrateSurface nature is key factor；In principle, any material that can be kept or keep its solid forms substantially, can be usedTo make substrate and cover plate, such as, the material that can act as substrate and cover plate can be monocrystalline silicon piece, quartz plate, sheet glass, heightPolymers such as dimethyl silicone polymer, polymethyl methacrylate, makrolon etc.；Certainly, the selection of substrate and the choosing of cover plateMaterial be able to can also be differed with identical；From the point of view of material consumption, manufacture difficulty and application popularization prospect etc. etc., theseNot small difference, the especially selection of that substrate between material be present, have a great influence.
In various substrate making materials, dimethyl silicone polymer, i.e. PDMS, comparatively very easily shaping, at thisIt is extremely simple that conduit is made on the substrate of sample, and the lower cost for material, substrate is made with the polydimethyl siloxane material,Conduit is being suppressed or etched thereon, and is being engaged with the cover plate that the cheap material such as glass or polypropylene or other plastic sheets makes,It is a kind of more satisfactory selection seemingly；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 asIt is extremely easy to popularize, promotes.
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 strongStrong hydrophobic material, conduit is built on this material, if without being operated for the modified of the conduit surface, then, it is wholeAfter body assembling is completed, that is, after covering cover plate, because the conduit its inner surface in structure occupies most fluid courseInner surface, then, its strong hydrophobic property of the PDMS conduits inner surface, is deciding factor, it can be similar to waterThe fine liquid stream of polar liquid of solution by becoming very difficult, its flow resistance is big, or even in general Micropump is all difficult toPromote, certainly, if cover plate also selects to use the PDMS material, then, problem is substantially identical, similar；Therefore, existingAmong having technology, modification is modified particular for the conduit inner surface on the PDMS material, is necessary operation；So, thisThe individual modified operation for PDMS conduit inner surfaces is pretty troublesomeThat falls nor this problem, forms serious technical puzzlement,It is another problem：PDMS polymer molecules inside its body phase of this PDMS material substrate have to be diffused to the surface, moves automaticallyThe characteristic of shifting, the characteristic that this substrate body phase inside PDMS polymer molecules are diffused to the surface, migrated automatically, will cause by tableState after the modification of its inner surface of that conduit of face modifying and decorating can not maintain the sufficiently long time, and that is through surfaceHolding time for conduit its inner surface state after modification is substantially only sufficient to the time needs for completing laboratory internal test experiments；In other words, by surface is modified or the PDMS conduit inner surfaces of surface modification, after it is modified or institute's shape after modification is saidInto surface state can not be lasting, but soon automatically tend to or say become again surface be modified before surface state, compared withThe strong hydrophobic surface state of that script is returned in the short time, then, just think, such micro-fluidic chip can be bigAmount making, mass storage, it is widely popularized, answer is it is obvious that is, impossible.Conduit on this PDMS material, does not doIf surface modification, it can not pump and pass through similar to the fine liquid stream of polar solvent of the aqueous solution, chip also cannot just use；And such asFruit has been cooked surface modification, can not persistently keep its state after modifying 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 surface-active is added in sample to be tested solutionAgent, so as to which 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 sample solutionIn tested composition occur combination so that tested composition can not be detected normally or it is detected ratio and dubiously droppedIt is low；The surfactant is possibly even fully wrapped around by tested composition with micelle form so that tested composition can not be detected orIdentification；Therefore, this 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 can be releasedInner surface decorating state can not persistently, chip can not largely make, largely lay in and then be widely popularized and such a make this areaThe puzzlement that numerous professionals 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 hydrophobicMaterial surface and also have another problem, that is, this strong hydrophobic PDMS surfaces can adsorb large biological molecule, andAnd these adsorbed large biological molecules can also further depression further on PDMS surfaces, gradually fall into it is gradually deep, untilIt is heavy to be trapped within the body phase of PDMS substrates, in fact, this process, partly it is also due to PDMS material body phase interior polymerMolecule, which has, to be diffused to the surface, caused by travel motion；Such case, it can also be explained from another angle, i.e. continue notDisconnectedly from inside PDMS body phases to those polymer molecules of its diffusion into the surface, migration, its result moved, be little by little by thatIt has been involved in a bit by the large biological molecule of adsorption within the body phase of PDMS substrates, briefly, these adsorbed biologiesMacromolecular is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up the phenomenon of large biological molecule, its instituteCaused by influence, 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 biological big pointSon, so, as the large biological molecule of experiment test object, its disappearance will not stop because surface saturation is adsorbed, andIt is, it is constantly adsorbed, also constantly swallowed up.
On PDMS substrates, its body phase is constantly swallowed up and tests showing for associated biomolecule macromolecular in related experiment test processAs, it is to say that another kind, which is explained, substantial amounts of Minute pores in PDMS body phases be present, associated biomolecule macromolecular by after adsorption,Depression enters these Minute pores, and then is swallowed up；However, inventor thinks, those can allow the sky of miniature scaleQi leel 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 enterEnter, both difference on yardstick are huge, must not make sweeping generalizations.Explanation is bypassed, in any case, as dependence test analysis objectLarge biological molecule is adsorbed by PDMS substrate conduits inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is known objectiveExisting phenomenon.
, can be from containment PDMS surfaces pair in order to prevent swallow up effect of this PDMS substrate bodies relative to large biological moleculeThe absorption of large biological molecule addresses, and method is chemically modified modification aiming at the PDMS material surface, forFor PDMS is the situation of substrate material, modification exactly is chemically modified to the surface of described channel portion, by chemistryThe conduit inner surface of modification, can contain its absorption to large biological molecule, and then avoid large biological molecule quiltPDMS substrate body phases are swallowed up；But or that old problem, that is, the chemical modification on PDMS material surface is modified itSurface state afterwards can not persistently be kept, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface, migrate automaticallyProcess, can soon by that by surface chemical modification be modified conduit inner surface state become again script it is strong it is hydrophobic simultaneouslyAnd 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 substrates itsConduit inner surface 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 is enoughReach and contain the absorption process of the PDMS substrate conduit inner surfaces to large biological molecule for a long time, and then prevent PDMS substrate bodies relativeThe 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 this area numerous specially as another problem addressed aboveIndustry 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.ShouldAlso be present many year in problem, so far, also not yet properly settled.
Third, the polydimethyl siloxane material, namely PDMS material, the active force very little wherein between polymer molecule,The polymer molecule of low polymerization degree inside its body phase is in constantly into the dynamic process of surface migration, therefore, includes PDMSAfter 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 point for the chip of substrate materialSon moves about, pours into the surface and change, and the change of its pattern is somewhat similarly to Rheological Deformation, trickling deformation or creep and becomePattern caused by shape institute is possible changes；In the higher PDMS material of some low polymerization degree component ratios, this kind somewhat similarCan be showed in the phenomenon of Rheological Deformation, trickling deformation or the deformation of creep must be than more significant；This kind include PDMS substrates chip itsThe dynamic of inner-walls of duct surface topography after shaping changes property, namely the described property or stream that are somewhat similarly to Rheological DeformationDropping down the property of deformation or the property of the deformation of creep, the word of rheology one is applied mechanically in this case, integration, also referred to as the rheological equationm of state or rheology becomeShape property, the dynamic of corresponding inner-walls of duct face pattern change, and this case is also referred to as rheology；As described above, because of this kind of materialExpect certain Rheological Deformation property as described above that its own has, its manufactured goods actually can at leisure Rheological Deformation and causeThe manufactured goods total form generation changes；Influence caused by its Rheological Deformation property, it is to be not easy to feel in terms of macroscopic perspectiveExamine, still, for the very fine micro-fluidic chip of its internal structure, the rheological equationm of state can but cause bigger shadowRing, the fluid course namely the pipeline therefore, during pre-production formed, because the wall of its tube chamber of pipeline is mainIt is the wall of PDMS materials, so, the cross-sectional area of the pipeline actually can gradually become because of rheology over timeIt is small, stated differently, since the reason for the rheology, the natively finer pipeline that is flowed through for liquid sampleIts tube chamber the more can become the more narrow, in the case of flow time is sufficiently long, or even the pipeline tube chamber can partly close or allGround closes, and then make it that sample fluid course is thoroughly blocked, and closes with being even also not reaching to its luminal part of pipelineThe degree closed or fully closed, its fluid passage that can be provided of that pipeline narrow because of rheology, then, in pipelineIts inner surface originally be exactly strongly it is hydrophobic in the case of, aqueous sample liquid is just more difficult to by becoming more so because of rheologyNarrow micro-channel, then, the influence of substrate rheology how is resisted, maintains the basic framework of the pipeline not occur seriously to become for a long timeThe problem of shape, also, keep the long-term unobstructed of the pipeline is 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, solves totally aboveA series of problems addressed, also, the solution is applied to AIDS diagnosis multichannel micro-fluidic chip field, formedA kind of new AIDS diagnosis Multichannel device.
The present invention solves the technical problem by following scheme, and the device that the program provides is a kind of with hydrophobic material workFor the AIDS diagnosis device of its substrate of micro-fluidic chip, the structure of the device includes multichannel micro-fluidic chip, the miniflowThe 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 sheetThe channel structure via mould pressing process or etching technics formation, the substrate are contained in thing, that face towards the cover plate of the substrateAlso contain and be connected and pierce being formed via mould pressing process, etching technics or simple drilling technology for the substrate with the channel structureWindow structure, be bonded to each other the substrate that is installed together and the cover plate and be built into jointly containing pipeline configuration and therewithThe micro-fluidic chip of connected liquid pool structure, the locations of structures of the pipeline are located at the interface area that the substrate is bonded to each other with the cover plateDomain, 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 structure bit of the liquid poolPut, the quantity of the liquid pool is three, and its locations of structures of two liquid pools therein is located at the sample introduction end of the micro-fluidic chip, remainingThe liquid flowing in its chip when being located at the actual sample introduction test of the micro-fluidic chip of liquid pool its locations of structures terminal, the endEnd is located remotely from each other with the sample introduction end, and one end of the pipeline is via the manifold shape turnout for being similarly positioned in the interface zone being bonded to each otherRespectively with the Liang Ge liquid pools UNICOM positioned at sample introduction end, the other end of the pipeline and the terminal positioned at the micro-fluidic chipA remaining liquid pool UNICOM, and, the working electrode that is sequentially respectively installed in the pipeline on diverse location and to electricityPole and reference electrode, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded AIDSThe 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 fourLateral 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 institute respectivelyState in four laterals, and, the specific antibody in its top layer gold size sensitivity membrane structure of four working electrodes is respectivelyThe four kinds of aids antibody materials that can be specifically bound to AIDS antigen, four kinds of antibody materials are aids antibody respectivelyP24 and aids antibody gp41 and aids antibody gp120 and aids antibody gp36, its material of the working electrodeIt is gold material or thermal decomposition conducting polymer material, sheet or thread is presented in its pattern of the working electrode, and emphasis is the baseIts material of piece is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, the surface of the primary form itsThe surface of the primary form for the material for not passing through any surface chemical modification or any surface chemical modification is intended to refer to,The structure of the device also includes hinge-type fixture, and hinge-type fixture its appearance profile likeness in form hinge, the hinge-type fixture is by phaseMutually hinged two hinges and through a fastening screw of two hinges and one and the fastening screwThe nut of fastening and hand break loose manually that is used for matched somebody with somebody and connected together with the fastening screw is formed, and the two of the hinge-type fixtureIndividual hinge is respectively mutually drawn close with its tip and clamps the micro-fluidic chip, and the tip is positioned at the neighbouring institute of the micro-fluidic chipState in the locations of structures of terminal, fixation attached in a hinge at least in and is equiped with miniature ultrasonic transducer units,And higher-order of oscillation electric signal transmission cable, one end of the higher-order of oscillation electric signal transmission cable and the miniature ultrasonic transducingDevice links together；The hinge-type fixture provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units itsMajor function is in the actual sample introduction test of micro-fluidic chip, utilizes the sample introduction end and the terminal and the miniature ultrasonicDifference in the distance between transducer installation position difference and its ultrasonic intensity experienced, enter described in induced synthesisDifference between its interfacial tension of sample end and the terminal its interfacial tension, the interfacial tension between the micro-fluidic chip both endsDifference can form pressure gap between the both ends of the micro-fluidic chip, and the pressure gap can drive sample solution to the endEnd flowing；It is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include with it to the strong suction of ultrasonic waveThe property of receipts, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip terminal to limited between the sample introduction endShort distance within realize the rapid decrement of ultrasonic intensity；And many silica dioxide granules, many silicaIt is particles filled in the pipeline, its tube chamber of the pipeline is filled up by many silica dioxide granules, the silicaFor its particle size range of grain between 10 microns and 200 microns, the silica dioxide granule is silica crystalline particles, silicaGlass particle or silica composition its inorganic glass particles of shared percentage by weight more than 80%.
Its particle diameter of the silica dioxide granule can be that the basis between 10 microns and 200 microns is actually needed and anyThe particle diameter specified, 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, lengthCube shape particle 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-mentionedThe 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.
Described miniature ultrasonic transducer units can certainly be all installed in two hinges；But only install oneIndividual miniature ultrasonic transducer units are dealt with enough to be used.
The word of hinge one art-recognized meanings of itself are known.
The gold size sensitive membrane is to be sufficiently mixed chitosan gold size solution and AIDS specific antibody solution uniformly, is usedPoint sample instrument point sample is coated on specified structure position, and forms its drying and forming-film.AIDS in the gold size sensitive membrane is specialHeterogenetic antibody is the aids antibody of horseradish peroxidase or glucose oxidase mark, and the gold size sensitive membrane has been wrappedContaining complementary medium therein is introduced for the above-mentioned each AIDS specific antibody of fixation, the complementary medium such as shell gathersSugar, cellulose acetate, gelatin be therein a kind of or their mixture.
The pipeline and the lateral and the manifold shape turnout in the microfluidic chip structure, in itFootpath size may each be arbitrarily selected size, still, for as far as possible less with prepare liquid sample and reduction reagent loss etc.Consideration, the pipeline and the lateral and the manifold shape turnout preferably select capillary level passage, instituteThe passage for stating capillary level implies that its internal diameter passage suitable with the internal diameter of the capillary on ordinary meaning.The capillary is in itThe shape of cross section of portion's passage can be arbitrary shape, and the shape of cross section is for example circular, oval, square, rectangle, barShape, naturally it is also possible to it is the linear of bending arbitrarily be present, also, the interior shape of the capillary is with the extension of pipeline,The shape of cross section of different parts can also allow to be different shapes.Only for the word of capillary one, its art-recognized meanings is publicKnow.
What is be related in structure is the electrode of microsize to electrode and reference electrode, and its electrode shape, which may each be, appointsThe selected shape of meaning, the arbitrarily selected shape such as square piece shape, rectangular patch, strip or round sheet etc..It is describedArt-recognized meanings to electrode 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 capsule for transitional liquid storageShape constructs, and its shape of the inner chamber of each liquid pool may each be arbitrarily selected shape, and the cavity shape is for example roundCylindrical cavity shape, square column type cavity-like, oblong cavity shape or spherical hollow space shape etc..The size of the liquid pool can be anySelected size, still, in order to as far as possible less with prepare liquid sample and reduce reagent loss, the liquid pool be preferably capable of withThe liquid pool of the microminiature of capillary matching.
Only for professional of the word of ultrasonic transducer one art-recognized meanings of itself for ultrasonic technology field,It is known.
Various sizes, variously-shaped ultrasonic transducer are commercially available；Its size of commercially available miniature ultrasonic transducer unitsIt may diminish to the magnitude only calculated with millimeter.
Only with regard to miniature ultrasonic transducer units its technique for fixing on general industry application solid body surface itsIt is known general technology for the professional in ultrasonic technology field for body.This case is not to this expansion superfluous words.
Only with regard to naked PDMS substrates itself conduit molding or lithographic technique for, be open-and-shut known technology；Similarly, 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.
The industrial products market of involved its all size of higher-order of oscillation electric signal transmission cable is on sale.
The structure of the device can also include higher-order of oscillation electric signal generator；The higher-order of oscillation electric signal transmission cableIts other end can be connected with the higher-order of oscillation electric signal generator.
The involved higher-order of oscillation electric signal generator technology of itself, come for the professional in ultrasonic technology fieldSay, be simple and known；The higher-order of oscillation electric signal generator can customize to ultrasonic instrument specialized factory.
The preferred scope of its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units is between 5 milliwatts and 9000 milliwattsBetween；The preferred scope of the frequency of its ultrasonic wave operationally launched of the miniature ultrasonic transducer units be between 100KHz withBetween 12MHz.
This case device can further include some annexes certainly, and the annex is such as multiple tracks electrochemical workstationDeng the art-recognized meanings of the multiple tracks electrochemical workstation are known.The each work being related in this case microfluidic chip structureElectrode and to electrode and reference electrode etc., special it can get lines crossed and the multiple tracks electrochemical workstation via corresponding respectivelyThe corresponding interface coupled.It is described that special to get lines crossed be for by each phase of each electrode and the multiple tracks electrochemical workstationAnswer the private cable that interface is coupled to each other.The micro-fluidic chip in this case device, its structure can also include micro-valve,The quantity of the micro-valve is unlimited, and according to being actually needed, the micro-valve can be installed in any need in the microfluidic chip structurePosition to be mounted；The word of micro-valve one is for the professional of micro fluidic chip technical field, the art-recognized meanings of itselfIt is known；The micro-valve itself manufacturing technology 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, still,It is recommending or say preferable its scope of the diameter between 0.1 micron to 2000 microns；The length of the working electrode canTo allow 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 1Micron is between 15000 microns.
It is installed in by spraying or point sample instrument point sample or the coating of other appropriate process described in the working electrode surface layerGold size sensitive membrane, its thicknesses of layers can allow be any setting treat sample measuring liquid occur electrical signals response thickness,It is however recommended to thickness preferable thickness is between 10 nanometers and 200 nanometers in other words.
The cover plate in chip structure, its material can allow to be any electrical insulating property material, such as：Polypropylene,Glass, polymethyl methacrylate, dimethyl silicone polymer, etc., in order to make smaller size of micro-fluidic chip, for example doInto the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and realized in the extremely short distance to ultrasonic waveExtremely fast decay, can preferably dimethyl silicone polymer be used as cover plate.Certainly, selected on large-sized micro-fluidic chipUsing dimethyl silicone polymer it is used as the cover plate, and this case technical scheme is allowed.
Positioned at the sample introduction end of the micro-fluidic chip two liquid pools its with positioned at the micro-fluidic chip terminal shouldAir line distance between a remaining liquid pool can be arbitrarily selected suitable distance, still, the distance as neededPreferred value is between 3 centimetres and 7 centimetres.
Described its thickness of cover plate and substrate can allow be any setting the thickness for being easy to assembling, the thickness of recommendation or sayPreferable thickness is between 1.0 millimeters and 5.0 millimeters.Less thickness is advantageous to save material.
The application method of this case micro-fluidic chip：
Itd is proposed first based on this case and the first public new liquid stream driving principle, its application of this case micro-fluidic chip are transportedAmong work, the new liquid stream driving method is determined completely without involving any additional Micropump.
The interfacial tension difference that this case is formed between micro-fluidic chip both ends caused by the ultrasonic wave,Driving liquid stream flows in the capillary channel of the four-way micro-fluidic chip, right respectively using four-way electrochemical analytical instrumentFour kinds of AIDS diagnosis antigens are detected.
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 ultrasonic wave driving, various AIDS antigen molecules are eachThe AIDS specific antibody for the corresponding horseradish peroxidase-labeled that gold size sensitive membrane embeds is caught on electrode surface in passageObtain.
2nd, the AIDS specific antibody of horseradish peroxidase-labeled is formed with the AIDS antigen in blood serum sample and exempted fromEpidemic disease compound.
3rd, using multi-channel electrochemical analyzer, the electron mediators such as catechol are added, it is above-mentioned using amperometric detectionCurent change caused by reaction, it is derived from the species and content of various analytes.
4th, result is subjected to comprehensive analysis, comprehensive diagnos is carried out to AIDS antigen.
It is an advantage of the invention that its close position positions the hinge-type folder in the terminal of the micro-fluidic chipTool, installation miniature ultrasonic transducer units are attached in the hinge-type fixture its hinge, are launched with the miniature ultrasonic transducer units lowThe ultrasonic wave of power, high-frequency band, meanwhile, using its strong absorbability to ultrasonic wave of dimethyl silicone polymer substrate,In shorter distance, it is, in from the terminal to the very short distance of only several centimeters yardsticks the sample introduction end,Reach the rapid decrement of ultrasonic intensity, the difference of the interfacial tension thereby caused at the both ends of the micro-fluidic chip,And then using the pressure gap between its both ends for being formed of the difference of the interfacial tension between the both ends, drive test liquidStream flows in the pipeline to the terminal direction.By this case liquid stream drive scheme, altogether dispense with traditionalThe equipment of Micropump etc；The micro-fluidic chip of Micropump is eliminated, its structure is more succinct, and its production process is less, its cost of manufactureLower, this kind of succinct structure is more beneficial for the low cost application for reaching micro-fluidic chip.
Its scheme of this case and by the pipeline except electrode installation take up space in addition to all clearance spaces wholeFilled up with the silica dioxide granule.Those are filled in the silica dioxide granule in pipeline, and this case is to its overall abbreviation titanium dioxideSilicon grain filler, its function include support, withstand the tube chamber of the pipeline its inwall so that the tube chamber of the pipeline is from because of instituteState rheology and narrow, prevent its reduced cross-sectional area of the tube chamber of the pipeline or even close, so as to maintain the base of the pipeline for a long timeSerious deformation does not occur for this framework.
Numerous silica dioxide granule entities are mutually banked up under this case framework together at random in the pipeline, the titanium dioxideThe function of silicon grain filler, also include certainly with the hydrophilic surface of its silica dioxide granule, utilize each adjacent titanium dioxideMutual formed complications are continuous and its inner surface that is mutually close together by water-wetted surface of silicon grain has hydrophilicThe space of property, in the hydrophilic fluid channel of the pipeline Inner Constitution network morphology, although this complications can be continuousAll the time the fluid channel with hydrophilic nmature, then, inside the tube chamber of the pipeline simultaneously and a plurality of miniflow depositedThe effect of its synthesis of road, integrated, cumulative superposition in other words, is the equal of the relatively fine hydrophilic capillary of caliberPassage；Its presence of the continuous hydrophilic fluid channel of the complications, the PDMS of the primary form is counteracted to a large extentThe flow resistance for liquid sample caused by the strong hydrophobic property in its surface of substrate；In other words, because of liquid sample, it is accounted forThe composition of maximum ratio is water, and liquid sample is substantially exactly the aqueous solution, therefore, the presence of the silica dioxide granule filler,Can significantly overcome with the unmodified PDMS substrates of hydrophobic property and surface its to the aqueous solution it is incompatible, repel andBarrier effect, so 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 described twoThe rigid of silicon oxide particle filler ties up, the table of part hydrophobic property that still can also be exposed on lumen wall wallIts area of face is already dramatically reduced, and the area on its still exposed part surface for belonging to hydrophobic property is muchLess than the wall surface area of the hydrophobic property of the lumen wall wall in the presence of no silica dioxide granule filler；So, under this case framework, in its lumen of the pipeline, its remaining hydrophobic surface increases surface newly with hydrophilic part, and it is foldedAdding, comprehensive, summation, cumulative technique effect, is to form the surface for tending to be hydrophilic on the whole；In other words, in instituteState inside the tube chamber of pipeline, its interfacial tension between sample solution of the part surface of remaining hydrophobic property, plus parentThe part of aqueous nature increases its interfacial tension between sample solution of surface newly, that it is superimposed, comprehensive, summation, cumulativeTechnique effect, be so that its solid-liquid interfacial tension inside the tube chamber of the pipeline synergy level off to capillary glass tube itsInterfacial tension between tube chamber inner surface and sample solution.
Therefore, exist in the pipeline under this case framework for having stated silica dioxide granule filler, while rely on thisCase sample liquid stream drive scheme, it becomes possible to the driving to sample liquid stream is reached with relatively low ultrasonic power, with this case mechanismThe test liquid is driven to flow to the end flow.
On the other hand, due to its rheological equationms of state of the PDMS as substrate material, it is filled in the pipelineThe interior silica dioxide granule filler, can gradually by constantly Rheological Deformation and to arest neighbors free space promote PDMSThe pipeline wall of material more tightly wraps up, and this process eventually causes those part two for pressing close to the lumen wall wallSilicon oxide particle is stuck in original place or is stuck in original place or embedding in-situ silica dioxide granule and its embedded in original place, the partIncarceration is together, described due to this reason each other for the remaining silica dioxide granule banked up with being mutually closely packed togetherSilica dioxide granule filler just will not move in the pipeline easily, silica dioxide granule therein be also substantially all byOriginal place is locked in, 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 between 10 microns and 200 microns, compared toFor general organic molecule, general large biological molecule, the particle diameter of its particle of this case silica dioxide granule filler can be rated as hugeGreatly, 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 notMutually 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 materialsSurface is adsorbed, and can not more be swallowed up by the inner surface of the pipeline of PDMS materials, the huge titanium dioxide of this kind of particle diameterSilicon grain surely not be fully sunk in easily or sink to completely PDMS materials its among intrinsic many Minute pores, andBecause above it is stated that the rheology the reason for, the silica dioxide granule filler can be stuck in original place in other words it is embeddingIn 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, such asUpper described, under this case framework, rigid because of the silica dioxide granule filler ties up, on lumen wall wall still alsoIts area of the surface of part hydrophobic property that can be exposed is already dramatically reduced, and it is still exposed to belong to hydrophobicityThe area on the part surface of matter has been far smaller than the tube chamber in the presence of no silica dioxide granule fillerThe wall surface area of the hydrophobic property of inwall wall；The reason for because of the rheology, it is close to those dioxies of lumen wall wallSilicon carbide particle, little by little it is partially embedded into or is partly absorbed among the lumen wall wall, the dioxy among the stateSilicon carbide particle its towards the part surface of lumen wall wall with have occurred and that matching deformation the lumen wall wall it is tightClosely connected conjunction, then, the matching is deformed and closely pasted with silica particles having occurred and that in lumen wall wallThe part wall of conjunction actually can not contact with flowing through the sample solution of pipeline, that is to say, that the wall of this part isThrough suction-operated can not be produced to the large biological molecule therein that flows through the sample solution of pipeline and organic molecule.Based on above-mentionedMechanism, then because fill up the pipeline the silica dioxide granule filler its presence, its lumen wall wall of the pipelineThe overwhelming majority in face is tightly covered, and these described walls tightly covered can not be with flowing through the pipelineSample solution contact, only remaining fraction be not brought into close contact by its surface of silica dioxide granule, tightly coverHydrophobic surface may be contacted with sample solution, in other words, under this case framework, large biological molecule in sample solution and haveIts direct touch opportunity with hydrophobic PDMS walls of machine molecule is significantly reduced, thus, the lumen wall faces of PDMS materials itsThe adsorptive hindrance and interference of swallowing up of large biological molecule and organic molecule are greatly reduced；As described above, this case schemeBe advantageous to exclude or weaken 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 being directed to PDMS materialsInner surface of pipeline carry out chemical modification effect, functioning as its this respect of this case is by the pipeline of the PDMS materialsSurface by the surface modification of hydrophobic property into the surface of hydrophilic nmature, still, with general, usual PDMS surface chemical modificationsIts hydrophilic chemical decorative layer retention time is short, hydrophilic effect can not keep situation prolonged enough different, particle described in this caseHuge silica dioxide granule filler, its particle being stuck by rheology can not both be moved easily, its huge particle diameterParticle can not more be swallowed up easily by the inner-walls of duct of PDMS materials, and therefore, its from this respect is similar to surface chemical modificationTechnique effect on see, its technique effect of this case, be form it is permanent, can not erase, can not consume, not be corroded, not byHydrophilic surface reforming layer swallowing up, being not dissolved, it is just comparable to one kind in its correlation table of PDMS base materials in effectThe permanent hydrophilic modifying superficial layer built on face.
It is related that the technical scheme of this case has dissolved its application to dimethyl silicone polymer substrate addressed above totallyA series of technical barriers.Based on this case scheme, the very cheap polydimethyl siloxane material of this kind is just possible to micro- at thisIt is prepared by fluidic chip, production, using etc. field play bigger effect.
The miniature ultrasonic transducer units have been installed in fixation in the hinge-type fixture its hinge in this case structure, the structureA function of facilitating the device to disassemble is provided, in this way, the hinge-type fixture is together with miniature ultrasonic appended in its hingeTransducer just easily can be mutually disengaged with the micro-fluidic chip, then, the component that the part can freely depart from just can be goodProperty is re-used cyclically many times；The architectural feature is advantageous to save the use cost of the device.
Embodiment
In this case that Fig. 1 and Fig. 2 are shown embodiment, the structure of the device includes multichannel micro-fluidic chip, shouldThe structure of micro-fluidic chip includes being bonded to each other the substrate 22 and cover plate 21 of installing together, and the substrate 22 and cover plate 21 areWhat is formed via mould pressing process or etching technics contained in plate object or tablet, that face towards the cover plate 21 of the substrate 22Channel structure, the substrate 22 also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technicsOr the window structure that simple drilling technology is formed, it is bonded to each other the substrate 22 being installed together and is built into jointly with the cover plate 21Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the liquid pool are liquid pool 1, liquid pool 2, liquid pool10, the locations of structures of the pipeline is located at the interface zone that the substrate 22 is bonded to each other with the cover plate 21, and its side of the window is by thisCover plate 21 blocks and opposite side opens, and the locations of structures of the window is exactly the liquid pool 1, liquid pool 2, the locations of structures of liquid pool 10,The quantity of the liquid pool is three, and two liquid pools therein are that liquid pool 1 and liquid pool 2 its locations of structures are located at the micro-fluidic chipSample introduction end 23, a remaining liquid pool are its chip when liquid pool 10 its locations of structures is located at the micro-fluidic chip actual sample introduction testThe terminal 24 of interior liquid flowing, the terminal 24 are located remotely from each other with the sample introduction end 23, and one end of the pipeline is via being similarly positioned in the phaseThe manifold shape turnout 3 for the interface zone being mutually bonded respectively with the liquid pool 1 positioned at sample introduction end 23 and the UNICOM of liquid pool 2, the pipelineThe other end and the remaining liquid pool of the terminal 24 positioned at the micro-fluidic chip be the UNICOM of liquid pool 10, and, according toSequence is respectively installed in working electrode in the pipeline on diverse location and to electrode 8 and reference electrode 9, the working electrodeIt is made up of conductive electrode and the gold size sensitive membrane for having embedded AIDS specific antibody being attached on the conductive electrode,The construction of the pipeline is in parallel construction, and the pipeline in parallel construction is made up of four lateral parallel connections, the work electricityThe quantity of pole is four, and respectively in four laterals, this four work the installation position of four working electrodesElectrode is working electrode 5, working electrode 6, working electrode 12, working electrode 13 respectively, and, four its top layers of working electrodeSpecific antibody in gold size sensitivity membrane structure is the four kinds of aids antibody things that can be specifically bound to AIDS antigen respectivelyMatter, four kinds of antibody materials be respectively aids antibody p24 and aids antibody gp41 and aids antibody gp120 andAids antibody gp36, for specifically deploying, 5 be in its top layer gold size sensitivity membrane structure being installed in lateral 4Specific antibody material is AIDS specific antibody p24 working electrode, and 6 be its top layer gold being installed in lateral 7The working electrode that specific antibody material in glue sensitivity membrane structure is AIDS specific antibody gp41,12 be to be installed in branchThe work that the specific antibody material in its top layer gold size sensitivity membrane structure in pipeline 11 is AIDS specific antibody gp120Electrode, 13 be that specific antibody material in its top layer gold size sensitivity membrane structure being installed in lateral 14 is special for AIDSHeterogenetic antibody gp36 working electrode, the working electrode 5, working electrode 6, working electrode 12, working electrode 13 its material areGold material or thermal decomposition conducting polymer material, the working electrode 5, working electrode 6, working electrode 12, working electrode 13Sheet or thread is presented in its pattern, and emphasis is, its material of the substrate 22 is dimethyl silicone polymer material, its surface of substrate 22The surface of primary form, the surface of the primary form its be intended to refer to and do not pass through any surface chemical modification or any tableThe surface of the primary form of the material of face chemical modification, the structure of the device also include hinge-type fixture 15, hinge-type folderHas 15 its appearance profile likeness in form hinge, the hinge-type fixture 15 is by two hinges 20,25 for being mutually hinged and runs throughOne fastening screw 17 of two hinges 20,25 and one match with the fastening screw 17 and are socketed with the fastening screw 17Together be used to fasten manually and the nut 26 of hand break loose is formed, two hinges 20,25 of the hinge-type fixture 15 respectively withIts tip is mutually drawn close and clamps the micro-fluidic chip, and the tip is positioned at the neighbouring terminal 24 of the micro-fluidic chipAttach to fix in locations of structures, in a hinge at least in and be equiped with miniature ultrasonic transducer units, in this legend,It is to attach to install the miniature ultrasonic transducer units 19 in hinge 20, it is of course also possible to allow all to fill in two hinges 20,25If upper miniature ultrasonic transducer units, but, changed it is, in general, that attaching one miniature ultrasonic of installing only in a hingeEnergy device is just enough to deal with to be actually needed, and, higher-order of oscillation electric signal transmission cable 18, the higher-order of oscillation electric signal transmission cable18 one end links together with the miniature ultrasonic transducer units 19；The hinge-type fixture 15 provides one and facilitates the device to tear openThe function of solution；Its major function of the miniature ultrasonic transducer units 19 is in the actual sample introduction test of micro-fluidic chip, using describedThe distance between sample introduction end 23 and the terminal 24 and the installation position of miniature ultrasonic transducer units 19 difference and its feltThe difference on ultrasonic intensity being subject to, its interfacial tension of sample introduction end 23 described in induced synthesis and described its interfacial tension of terminal 24Between difference, the micro-fluidic chip both ends are that the interfacial tension difference between sample introduction end 23 and terminal 24 can be micro-fluidic at thisThe both ends of chip are to form pressure gap between sample introduction end 23 and terminal 24, and the pressure gap can drive sample solution to describedFlow in the direction of terminal 24；It is soft and have the substrate 22 of the dimethyl silicone polymer material of elasticity its function and also include with it to superThe property that sound wave absorbs strongly, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip, the terminal 24 arrives the sample introductionThe rapid decrement of ultrasonic intensity is realized within limited short distance between end 23；And many silica dioxide granules, shouldMany silica dioxide granules are filled in the pipeline, and its tube chamber of the pipeline is filled out by many silica dioxide granulesFull, for its particle size range of the silica dioxide granule between 10 microns and 200 microns, the silica dioxide granule is titanium dioxide silicon waferIts unorganic glass of shared percentage by weight more than 80% of state particle, silica glass granule or silica compositionGrain.
Arrow in legend indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, its tryThe flow direction of sample liquid stream.
The miniature ultrasonic for being depicted without attaching installing in Fig. 1 in the hinge-type fixture 15 and one hinge changesCan device 19 and higher-order of oscillation electric signal transmission cable 18；Also, the higher-order of oscillation telecommunication is depicted without in Fig. 1 and Fig. 2The associate member such as number generator and multiple tracks electrochemical workstation.
Involved hinge-type fixture 15 can be carried out simply cutting using commercially available hinge, changed a social system；The miniature ultrasonicTransducer 19 is commercially available；Commercially available miniature ultrasonic transducer units are various in style, can select to use as needed；This is miniature superAcoustic wave transducer 19 can also customize to ultrasonic transducer specialized factory.
Involved higher-order of oscillation electric signal transmission cable 18 is commercially available；Can also be special to ultrasonic transducer producer or cableIndustry producer customizes.
Involved higher-order of oscillation electric signal generator in the market has the product close to needs commercially available；It can also determine to relevant manufacturersSystem.
Each working electrode in this example structure and can be respectively via each special electricity to electrode and reference electrodeCable or say is being got lines crossed respectively with the corresponding cable interface of the multiple tracks electrochemical workstation as annex or saying interface connection of getting lines crossed.