CN106607109A - Cheap hydrophobic substrate-based chip device used for screening of common tumor markers - Google Patents

Abstract

The invention relates to a cheap hydrophobic substrate-based chip device used for screening of common tumor markers, belonging to the field of analytical test. PDMS, i.e., polydimethylsiloxane, has both advantages and problems when applied to preparation of a substrate for a universal micro-fluidic chip used for combined detection of dozens of tumor markers. Directed at the problems, the invention provides the cheap hydrophobic substrate-based chip device. According to main points of a technical scheme in the invention, PDMS with an original surface is selected and used as the substrate; a hinge-type clamp, additionally provided with a minisized ultrasonic transducer, is manually fastened and positioned at a position close to a sample liquid flow terminal of the micro-fluidic chip, and ultrasonic waves are used for reducing interfacial tension; meanwhile, in virtue of strong ultrasonic wave absorption capacity of PDMS, rapid and progressive decrease of ultrasonic intensity is realized in a short distance; thus, a difference of interfacial tension is formed at two ends of the chip and supplies a force for driving sample liquid flow to flow towards the terminal along a hydrophobic capillary channel, and the force is coupled and cooperated with the mechanical pumping force of a micropump included in the device.

Description

It is related to the common tumor markers examination chip apparatus of cheap hydrophobic substrate

Technical field

The present invention relates to a kind of common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate, belong to analysis test neck Domain.

Background technology

Tumor markerses (tumor marker, TM) are referred in the generation and breeding of tumor, produced by tumor cell itself It is raw or by body to tumor cell react and produce, reflect tumor exist and grow a class material, including protein, Hormone, enzyme (isozyme) and oncoprotein etc..Tumor markerses in chemical examination blood samples of patients or body fluid, can be in cancer screening Early discovery tumor, and observe the curative effect of oncotherapy and judge patient's prognosis.At present clinically conventional tumor markerses have： (1) alpha-fetoprotein (AFP) is the mark of the tumors such as primary hepatocarcinoma, carcinoma of testis, ovarian cancer；(2) carcinoembryonic antigen (CEA) is digestion The mark of the tumors such as system tumor, pulmonary carcinoma, breast carcinoma；(3) CA125 (CA125) is the mark of the tumors such as ovarian cancer； (4) CA153 (CA153) is the mark of the tumors such as breast carcinoma；(5) CA19-9 (CA19-9) is digestive system tumor Mark；(6) CA724 (CA724) is the mark of the tumors such as gastric cancer, ovarian cancer；(7) carbohydrate antigen 242 (CA242) For the mark of digestive system tumor；(8) CA50 (CA50) is the mark of the tumors such as digestive system tumor, breast carcinoma, pulmonary carcinoma Thing；(9) CYFRA21-1 (Cy211) is the mark of the tumors such as nonsmall-cell lung cancer；(10) neuronspecific enolase (NSE) For the mark of the tumors such as small cell lung cancer, neuroendocrine tumor；(11) prostate specific antigen (PSA) is carcinoma of prostate Tumor markerses；(12) human chorionic gonadotropin (HCG) is that ECC, trophoblastic tumor (choriocarcinoma, hydatidiform mole) etc. are swollen The mark of tumor；(13) Elityran (TG) is the mark of thyroid carcinoma；(14) ferritin (SF) is digestive system tumor, liver The mark of the tumors such as cancer, breast carcinoma, pulmonary carcinoma；(15) B2M (β 2-MG) is in chronic lymphocytic leukemia, lymph Raise in the patient body fluids such as tumor, myeloma, pulmonary carcinoma, thyroid carcinoma, nasopharyngeal carcinoma；(16) squamous cell antigen (SCC) be cervical cancer, The tumor markerses such as lung squamous cancer, the esophageal carcinoma.The tumor markerses overwhelming majority for clinically detecting at present is not only present in malignant tumor In, exist in benign tumor, embryonal tissue, even in normal structure.Therefore, tumor markerses have dynamic chek and multinomial Joint inspection is more valuable.So for numerous tumor markerses, clinically how to selectDifferent tumors understands some phases To special tumor markerses, such as CA153 often occurs in breast carcinoma；CEA often occurs in intestinal cancer, gastric cancer；CA19-9 is often occurred in Intestinal cancer, cancer of pancreas；CA125 often occurs in ovarian cancer etc..Clinician can be according to the different mark of different tumor examinations. Same tumor or different types of tumor can have one or more tumor markerses exceptions；Same tumor markerses can be in difference Tumor in occur.To improve the additive diagnostic value of tumor markerses and determining which kind of mark can be supervised as the follow-up after treatment Index is surveyed, tumor markerses joint-detection, the complementary tumor markerses group of several sensitivity of reasonable selection, specific performance can be carried out Into best of breed, joint-detection is carried out.In general the joint-detection of tumor markerses can improve the accuracy to diagnosing tumor.

Only with regard to Diagnostic Value of Several Serum Tumor Markers its joint-detection background of related itself general picture or overview for, may refer to Lower Chinese invention patent application case：CN200410041175.3、CN200510026780.8、CN200610040051.2、 CN200910064647.X。

Only for the microfluidic chip technology overall general picture of itself, famous micro-fluidic expert Mr. Lin Ping Cheng is may refer to soon Before the monograph " diagram Microfluid based Lab on a chip " that goes out, the monograph publishes via Science Press, and the monograph is for micro-fluidic Past of technology, now, and, vision of the future etc. aspect, suffer from detail, be deep into long discussion of detail.

So, the focal issue that this case that to have a talk below pays special attention to.

The basic framework of micro-fluidic chip, including the substrate for being etched with small fluid course and the cover plate for fitting together therewith, Small fluid course on the substrate, before assembling upper cover plate, it is apparent on see to be exactly some micro-channel, be when thereon After covering cover plate, just real closure forms the small fluid course, and the conduit inner surface of the micro-channel is together with surround this The part cover plate of micro-channel constitutes together described small fluid course；So, it is clear that this after assembling is completed is small Fluid course, the major part of its inner surface area is the inner surface area of that micro-channel, in other words, in the micro-channel The state or property on surface substantially determines the integrality or property of the small fluid course；Therefore say, this is built in base The inner surface state or inner surface property of the micro-channel on piece is key factor；In principle, it is any to keep or protecting substantially The material of its solid forms is held, can be used to make substrate and cover plate, such as, the material that can act as substrate and cover plate can be with It is monocrystalline silicon piece, piezoid, sheet glass, high polymer such as polydimethylsiloxane, polymethyl methacrylate, Merlon etc. Deng；Certainly, the selection of substrate can be with identical with the selection of cover plate, it is also possible to differs；From material consumption, manufacture difficulty and Application popularization prospect etc. aspect from the point of view of, there is not little difference between these materials, especially the selection of that substrate, affect compared with Greatly.

In various substrate making materials, polydimethylsiloxane, i.e. PDMS, comparatively very easy molding, so Substrate on make that micro-channel is extremely simple, and the lower cost for material makes substrate with the polydimethyl siloxane material, Micro-channel is suppressed or etched thereon, and the cover plate made with glass or the cheap material such as polypropylene or other plastic sheets is engaged, It is seemingly 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, makes extremely simple, seems and should also be as It is extremely easy to popularize, promotes.

But, thing is really not so simple.

First, this polydimethyl siloxane material, that is, the material that the letter PDMS that abridges is referred to, itself are a kind of strong Hydrophobic material, on this material micro-channel is built, if not carrying out the modified operation for the micro-channel surface, then, Integral installation is covered after cover plate with after completing, because of structure in the micro-channel its inner surface occupy most liquid circulation The inner surface in road, then, the PDMS micro-channel inner surface its strong hydrophobic property is deciding factor, and it can cause class Be similar to aqueous solution the fine liquid stream of polar liquid by becoming very difficult, its flow resistance is big, or even general Micropump is all It is difficult to promote, certainly, if cover plate also selects to use the PDMS material, then, problem is substantially identical, similar； Therefore, among prior art, modification is modified particular for the micro-channel inner surface on the PDMS material, is necessary Operation；So, this is pretty troublesome for the modified operation of PDMS micro-channel inner surfaciesThat falls nor this problem, structure It is another problem into serious technical puzzlement：PDMS polymer molecules tool inside this PDMS material substrate its body phase There is the characteristic for diffusing to the surface automatically, migrating, this substrate body phase inside PDMS polymer molecules are diffused to the surface, moved automatically The characteristic of shifting, by cause through that micro-channel of surface modifying and decorating its inner surface it is modified after state can not maintain foot The enough long time, being held time of the micro-channel after that is surface-modified its inner surface state be substantially only sufficient to, and to complete experiment indoor The time of portion's test experiments needs；In other words, PDMS micro-channel inner surfacies of or surface modification modified through surface, its After modified or say the apparent condition formed after modification can not be lasting, but soon automatically tend to or say become surface again and change Apparent condition before property, returns to the strong hydrophobic apparent condition of that script in the shorter time, then, just think, Such micro-fluidic chip can make in a large number, mass storage, be widely popularized, and answer is it is obvious that is, impossible. Micro-channel on this PDMS material, does not do if surface modification, cannot pump similar to the fine liquid stream of polar solvent of aqueous solution Send and pass through, chip also just cannot be used；And if having done surface modification, the state after its modification cannot be persistently kept again, also Being equally cannot popularization and application.

So, how to accomplish that substrate can either be made using cheap PDMS material, and table in the micro-channel can be released Face decorating state cannot persistently, chip cannot in a large number make, deposit and then be widely popularized and such a make this area numerous specially in a large number The puzzlement that industry 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 days in the highly difficult problem, so far, not yet properly settled.

Second, the PDMS material of non-surface modification, above it is stated that, its surface is strongly hydrophobic, this strong hydrophobic Material surface and also another problem, that is, this strong hydrophobic PDMS surfaces can adsorb biomacromolecule, and And, the depression that these adsorbed biomacromolecules can also be further further on PDMS surfaces gradually falls into 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 have diffuse to the surface, travel motion is caused；Such case, it is also possible to explain from another angle, i.e. Continuously from inside PDMS body phases to its diffusion into the surface, migration those polymer molecules, its motion result, be by Gradually those are involved within the body phase of PDMS substrates by the biomacromolecule of surface adsorption, briefly, these are inhaled Attached biomacromolecule is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up biomacromolecule Phenomenon, the impact caused by it necessarily causes to be related to the severe deviations of all kinds of test data of experiment of biomacromolecule.

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

With regard to PDMS substrates in related experiment test process its body phase constantly swallow up test associated biomolecule macromole phenomenon, separately It is to say that one kind is explained, there are substantial amounts of Minute pores in PDMS body phases, associated biomolecule macromole by after surface adsorption, depression Into these Minute pores, and then swallowed up；However, inventor thinks, those can allow the air point of miniature scale Son clamp-ons the Minute pores therebetween, and being not equal to them also can directly allow that the biomacromolecule of relative large scale is entered, and two Person's difference on yardstick is huge, must not make sweeping generalizations.Explanation is bypassed, in any case, as dependence test the biology of object is analyzed Macromole is adsorbed by PDMS substrate micro-channel inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is known objective The phenomenon of presence.

In order to prevent this PDMS substrate bodies relative to the effect of swallowing up of biomacromolecule, can be from containment PDMS surfaces opposite The absorption of thing macromole addressing, method be aiming at the PDMS material surface be chemically modified it is modified, for For PDMS is for the situation of substrate material, exactly modified, Jing Guohua is chemically modified to the surface of described micro-channel part The micro-channel inner surface of modification is learned, its absorption to biomacromolecule can be contained, and then avoid biomacromolecule quilt PDMS substrate body phases are swallowed up；But, or that old problem, that is, the chemical modification on PDMS material surface changes Apparent condition after property cannot persistently keep, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface automatically, The process of migration, soon can become that again script strong and dredge through the micro-channel inner surface state that surface chemical modification is modified The state of water and strong adsorption biomacromolecule, in other words, no matter how professionals in the field turn from side to side, the PDMS Substrate its micro-channel 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, can reach again Growth stage contains the absorption process of the PDMS substrate micro-channel inner surfacies to biomacromolecule, and then prevents PDMS substrate body phases The effect of swallowing up to biomacromolecule so that related chip manufactured goods are able to maintain that a prolonged enough, rational shelf-life, It is exactly a very thorny difficult problem.The difficult problem makes the numerous specialties in this area as another difficult problem addressed above, equally Personnel are entangled with for a long time, perplex, and the difficult problem is equally the highly difficult problem that its obvious technology barrier can not despise.The hardly possible Also be present many days in topic, so far, also not yet properly settled.

Third, as described above, the PDMS micro-channel inner surfacies are strongly hydrophobic, and targetedly surface chemical modification or come to the surface Learn modification to be difficult to again persistently, therefore, actually can only its surface is modified or surface modification after apparent condition still belong to effectively short It is used within phase；If have passed through that ofer short duration expiry date, and still by force if use, due to surface shape State already again near hydrophobic state, drives test liquid stream then to certainly exist than larger flow resistance using usual Micropump, So, test liquid just can only be forced by increasing Micropump pump power and pumping pressure to flow to target direction flowing, as described above, This PDMS material is soft, and with too high, machinery pumping pressure test liquid stream is pumped, and it enters will to cause the substrate Sample end includes that the micro-channel of sample introduction end near zone occurs bubbling, expanded, distortion, deformation, also, this high-pressure situations Under, the micro-channel and its periphery in the sample introduction end and its near zone is also susceptible to the stripping between substrate and cover plate, this feelings Under shape, sample solution everywhere flows over the crack of the appearance between the substrate formed into after the stripping and cover plate, this reality Border causes the damage of the micro-fluidic chip；Certainly, if the surface is modified or surface modification is not in place if originally, also result in There is said circumstances within of short duration usual effect duration；It is above-mentioned in the case where liquid stream driving is carried out using additional Micropump merely The problem is present all the time.If as described above, do not did that any surface is modified completely or the pre action such as surface modification, So, the above-mentioned problem will be even more serious, even without occur sample introduction end and its micro-channel bubbling described near zone, it is expanded, Between distortion, deformation and substrate and cover plate the problems such as peel off, merely because the flow resistance is excessive, using high pressure Micropump also not Test liquid stream must be driven to march forward towards terminal side.

The content of the invention

The technical problem to be solved is to provide a package solution, while solving three for addressing above The a series of difficult problem for actually mutually involving together of aspect, also, it is a kind of new that the solution is applied to into structure Crowd's health check-up is blanket can be directed to ten kinds carry out simultaneously examination than more typical primary tumor mark, while detection is micro- Fluidic chip device.

The present invention solves the technical problem by following scheme, and the device that the program is provided is that one kind is related to cheap hydrophobic substrate Common tumor markers examination chip apparatus, program main points are, the structure of the device includes micro-fluidic chip, the miniflow The structure of control chip includes the substrate installed together bonded to each other and cover plate, and the substrate and cover plate are plate object or lamellar The channel structure formed via mould pressing process or etching technics is contained in thing, that face towards the cover plate of the substrate, bonded to each other The substrate being installed together has been built into the micro-fluidic chip containing pipeline configuration, the locations of structures of the pipeline jointly with the cover plate Positioned at the substrate and cover plate juncture area bonded to each other, the two ends of the pipeline respectively with the sample introduction end of the micro-fluidic chip and Terminal connects, and the sample introduction end is the injection end of the micro-fluidic chip sample solution, and the terminal is that the actual sample introduction of the micro-fluidic chip is surveyed The terminal that sample solution flows in its chip during examination, the terminal is located remotely from each other with the sample introduction end, between the terminal and the sample introduction end Distance between 3 centimetres and 10 centimetres, on diverse location in the pipeline in order or backward be equiped with working electrode and To electrode and reference electrode, the order refers to the reference electrode its locations of structures closer to the terminal location, described Backward refers to the reference electrode locations of structures closer to the sample introduction end position, the working electrode by conductive electrode and The gold size sensitive membrane for having embedded tumor markerses antibody being attached on the conductive electrode is constituted, and the construction of the pipeline is presented in parallel Construction, the pipeline in parallel construction is made up of 15 lateral parallel connections, the pipeline that parallel construction is presented its Appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is 15, the installing of 15 working electrodes Position is located in 15 laterals respectively, and, in the sensitive membrane structure of its top layer gold size of 15 working electrodes Tumor markerses antibody is respectively the 15 kinds of tumor markerses antibody materials that can be specifically bound to tumor markerses antigen, and this ten Five kinds of antibody materials be respectively tumor markerses antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, the antigen is the antigen of broad sense, described Antibody is the antibody of broad sense, and the working electrode its material is that argent material, gold material, carbon material or thermal decomposition are conductive Macromolecule material, the working electrode its pattern is presented column, lamellar or thread, and the substrate its material is polydimethylsiloxane Material, its surface of the substrate is the surface of primary form, the surface of the primary form its be intended to refer to not through any surface The surface of the primary form of the material of chemical modification or any surface chemical modification, the structure of the device is also pressed from both sides including hinge-type Tool, the hinge-type fixture its appearance profile likeness in form hinge, the hinge-type fixture is by two hinges being mutually hinged and passes through Wear a fastening screw and the use for matching with the fastening screw and connecting together with the fastening screw of two hinges Constitute in the nut of fastening and hand break loose manually, two hinges of the hinge-type fixture are respectively mutually drawn close with its tip and clamp this Micro-fluidic chip, the tip is positioned in the locations of structures of neighbouring described terminal of the micro-fluidic chip, at least in one Fixation is attached in the individual hinge and is equiped with miniature ultrasonic transducer units, and, higher-order of oscillation electric signal transmission cable, the high frequency One end of vibration electric signal transmission cable links together with the miniature ultrasonic transducer units；The hinge-type fixture provides a side Just the function that the device is disassembled；The miniature ultrasonic transducer units its major functions is the profit when the actual sample introduction of micro-fluidic chip is tested The ultrasound wave launched with it is made reducing the interfacial tension between sample solution and the inwall of its inner passage of micro-fluidic chip It can be compatible, also, using between the sample introduction end and the terminal and the miniature ultrasonic transducer units installation position away from Deviation is different and its ultrasonic intensity for being experienced on difference, its interfacial tension of sample introduction end described in induced synthesis and the terminal Difference between its interfacial tension, the interfacial tension difference between the micro-fluidic chip two ends can the micro-fluidic chip this two Pressure gap is formed between end, the pressure gap can drive sample solution to the end flow；The miniature ultrasonic transducer units its Function also include the ultrasound wave launched with it check biomacromolecule contained in sample its in the micro-fluidic chip inside it Absorption on channel inner surface, and then check the substrate of the polydimethylsiloxane material its body phase the biomacromolecule is swallowed up Effect；It is soft and have the substrate of the polydimethylsiloxane material of elasticity its function and include with its property to the strong absorption of ultrasound wave Matter, is absorbed strongly to ultrasound wave, and thereby in the micro-fluidic chip terminal to the limited short distance between the sample introduction end Within realize the rapid decrement of ultrasonic intensity；And, Micropump, the Micropump is connected with the sample introduction end；The function of the Micropump is, Interfacial tension between the inwall of the sample solution with its inner passage of micro-fluidic chip is reduced by the ul-trasonic irradiation, phase Between under the increased precondition of the compatibility, come and the two ends of ultrasound wave induction between with the mechanicalness of Micropump pumping strength Interfacial tension difference its driving force brought support mutually, adjust mutually, be mutually coupled, converged in the way of Collaboration Being polymerized to one drives test liquid to flow to the strength of the terminal direction flowing.

The tumor markerses antibody is the antibody of broad sense, and the antibody of the broad sense is referred to possess antibody function or is functionally similar to Immune complex or immune compound can be combined and be formed in antibody with the involved tumor markerses of various corresponding clinics The material of the analog of thing；The tumor markerses antigen is the antigen of broad sense, and the antigen of the broad sense is referred to can be using corresponding Antibody or be functionally similar to antibody material carry out enzyme mark detection it is various clinic involved by the tumor-marker for needing to differentiate, detect Thing.

Described miniature ultrasonic transducer units can certainly be all installed in two hinges；But, only installing one is micro- Type ultrasonic transducer is dealt with enough and is used.

The word of hinge one art-recognized meanings of itself are known.

The word of the 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.

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

The gold size sensitive membrane is to be sufficiently mixed shitosan gold size solution and tumor markerses antibody-solutions uniformly, uses point sample instrument Point sample coats specified structure position, and forms its drying and forming-film.Tumor markerses antibody in the gold size sensitive membrane is equal For horseradish peroxidase or the tumor markerses antibody of glucose oxidase labelling, the gold size sensitive membrane is included as fixing Above-mentioned each tumor markerses antibody and introduce complementary medium therein, the complementary medium for example shitosan, cellulose acetate, Gelatin is therein a kind of or their mixture.

The pipeline in the microfluidic chip structure includes the lateral, and its internal diameter size may each be arbitrarily selected Size, but, for prepare liquid sample and the consideration of the aspects such as reagent loss is reduced less as far as possible, the pipeline is including described The passage of the preferred capillary level of lateral, the passage of the capillary level implies that the interior of the capillary tube on its internal diameter and ordinary meaning The suitable passage in footpath.The shape of cross section of its inner passage of capillary tube can be arbitrary shape, the shape of cross section example Such as circular, oval, square, rectangle, bar shaped, naturally it is also possible to be arbitrarily the presence of the linear of bending, also, the hair With the extension of pipeline, the shape of cross section of different parts can also allow to be different shapes the interior shape of tubule.Only with regard to hair For the word of tubule one, its art-recognized meanings is known.

What is be related in structure is minute sized electrode to electrode and reference electrode, and its electrode shape may each be arbitrarily choosing Fixed shape, the arbitrarily selected shape such as column, lamellar, strip or thread etc..It is described to electrode and the ginseng It is known than the art-recognized meanings of the electrode vocabulary of itself.

It is public for the word of ultrasonic transducer one art-recognized meanings of itself of professional only with regard to to(for) ultrasonic technology field Know.

Various sizes, variously-shaped ultrasonic transducer are commercially available；Commercially available miniature ultrasonic transducer units its sizes can be with It is little to only with millimeter calculate magnitude.

Only with regard to miniature ultrasonic transducer units its technique for fixing on general industry application solid body surface itself and Speech, is known general technology for the professional in ultrasonic technology field.This case is not to this expansion superfluous words.

Only with regard to naked PDMS substrates itself micro-channel molding or lithographic technique for, be open-and-shut known technology.

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；Its is another for the higher-order of oscillation electric signal transmission cable One 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, for the professional in ultrasonic technology field, be It is simple and known；The higher-order of oscillation electric signal generator can be customized to ultrasonic instrument specialized factory.

The preferred scope of the miniature ultrasonic transducer units its specified ultrasonic emitting power be between 5 milliwatts and 5000 milliwatts it Between；The preferred scope of the frequency of the miniature ultrasonic transducer units its ultrasound wave operationally launched be between 100KHz with Between 12MHz.

This case device can further include some adnexaes, the adnexa 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 To electrode and reference electrode etc., can respectively via corresponding special the corresponding interface got lines crossed with the multiple tracks electrochemical workstation Coupled.It is described that special to get lines crossed be for each electrode is carried out into phase with each the corresponding interface of the multiple tracks electrochemical workstation 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 Amount is not limited, and according to actual needs, the micro-valve can be installed in any need position to be mounted in the microfluidic chip structure； For the professional of micro fluidic chip technical field, the art-recognized meanings of itself are known to the word of micro-valve one；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 for using, it is however recommended to Or say preferred 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 for using, it is however recommended to or to say the preferred length its scope be at 1 micron To between 15000 microns.

The gold size for being installed in the working electrode surface layer by spraying or point sample instrument point sample or other appropriate process coatings is quick Sense film, its thicknesses of layers can allow the thickness for being the sample measuring liquid the treated generation electrical signals response of any setting, but, push away In other words conj.or perhaps preferred thickness is between 10 nanometers and 200 nanometers to the thickness recommended.

The cover plate in chip structure, its material can allow to be any electrical insulating property material, for example：Polypropylene, glass Glass, polymethyl methacrylate, polydimethylsiloxane, etc., in order to make smaller size of micro-fluidic chip, such as do Into the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and realize to ultrasound wave in the extremely short distance Extremely fast decay, it may be preferred to polydimethylsiloxane is used as cover plate.Certainly, select on large-sized micro-fluidic chip It is used as the cover plate using polydimethylsiloxane, is also that this case technical scheme is allowed.

The cover plate and substrate its thickness can allow the thickness for being easy to assemble for being any setting, the thickness of recommendation or say preferably Thickness be between 1.0 millimeters and 5.0 millimeters.Less thickness is conducive to material-saving.

The using method of this case micro-fluidic chip：

This case drives liquid stream to flow in the capillary channel of the ten Five-channels micro-fluidic chip with Dual Drive coupling operating mode It is dynamic, respectively joint-detection is carried out to 15 kinds of tumor markerses using multi-channel electrochemical analyzer device.

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

1st, blood serum sample liquid is added in micro-pipe road, in the case where Dual Drive coupling operating mode drives, various tumor markerses Antigen molecule is by the tumor-marker of the corresponding horseradish peroxidase-labeled of gold size sensitive membrane embedding on electrode surface in each passage Thing specific antibody is captured.

2nd, the tumor markerses antigen in the tumor markerses specific antibody and blood serum sample of horseradish peroxidase-labeled is formed Immune complex.

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

4th, result is carried out into comprehensive analysis, comprehensive diagnos is carried out to tumor markerses antigen.

It is an advantage of the invention that its close position positions the hinge-type fixture in the terminal of the micro-fluidic chip, with The miniature ultrasonic transducer units installed, the low-power launched using it, high-frequency band are attached in the hinge-type fixture its hinge Ultrasound wave so that without surface chemical modification or surface chemical modification the strong hydrophobic micro-fluidic chip internal pipeline its The compatibility between tube wall and test object aqueous solution is significantly increased, and this is test liquid stream by possible there is provided a reality Property；Meanwhile, using its strong absorbability to ultrasound wave of polydimethylsiloxane substrate, in shorter distance, also Be, from the terminal to the very short distance of the only several centimeters yardstick the sample introduction end in, reach the quick of ultrasonic intensity Successively decrease, the difference of the interfacial tension is thereby caused at the two ends of the micro-fluidic chip, the interfacial tension between the two ends Difference can cause a kind of driving force, and because of interfacial tension difference, its function of caused driving force is to drive test liquid stream to exist to this kind Originally to terminal direction flowing in strong hydrophobic capillary channel；And the simultaneous Micropump in structure, its function It is that the interfacial tension between the inwall of the sample solution and its inner passage of micro-fluidic chip is dropped by the ul-trasonic irradiation Under the precondition that low, the alternate compatibility increases, described two with the ultrasound wave induction are carried out with the mechanicalness pumping strength of the Micropump Interfacial tension difference between end its driving force brought is supported mutually, adjusts mutually, is mutually coupled, with Collaboration Mode pools one and drives test liquid to flow to the strength of the terminal direction flowing；The presence of the Micropump so that the miniature ultrasonic Wave transducer its ultrasonic emitting intensity can be allowed to appropriate reduction, this in detection object containing ultrasound wave sensitive composition Situation is particularly suitable for；And due to the presence of the ultrasonic transducer and its radiated ultrasound wave, it is possible to increase the alternate compatibility, drop Low interfacial tension, and the two ends interfacial tensions difference special driving force that it is brought is provided, then, in this case, Test liquid stream its flow resistance in the micro-channel is greatly reduced, and correspondingly, the Micropump its running resistance is greatly reduced, so, The Micropump just can be to carry out the pumping work for the sample solution than relatively low pumping pressure, due to mechanical pumping Pressure is greatly reduced, therefore, in such a situation, just it is not susceptible to because sample introduction terminal tool pumping pressure is excessive and caused Between the micro-channel bubbling of the sample introduction end and its near zone, expansion, deformation, distortion and the region substrate and cover plate The problems such as stripping etc.；The scheme of this case Dual Drive coupling running and increased manipulation for the sample fluid flow action Property, can allow for coming using the multiple indexs of ultrasonic intensity, ultrasonic frequency, Micropump pump power, Micropump pumping pressure etc. It is many that flow rate, flowing action for the flowing includes that flow forward or time-out flowing or acceleration flowing etc. flowing action are carried out Parameter is accurately manipulated；By the scheme of this case Dual Drive coupling running, entirely without must be to the base of the polydimethylsiloxane material Piece its micro-channel etc. relevant surfaces carry out any surface chemical modification or surface chemical modification, have altogether dispensed with the surface chemistry Modification or the laborious procedures of surface chemical modification；On the other hand, the ultrasound wave of the low-power, high-frequency band, additionally it is possible to containment examination Absorption of the biomacromolecule in sample on literalness naked its inner surface of pipeline of polydimethylsiloxane substrate, and then contain The swallow up effect of its body phase of polydimethylsiloxane substrate to the biomacromolecule；The antigen, antibody and antigen with it is anti- The Reversible binding thing of body is all belonging to the type of described biomacromolecule certainly；Due to described adsorption and described gulp down Do not act on and effectively being contained, therefore, dependence test result will be better able to objectively reflect practical situation；The low-power, height Again and again the effect of section ultrasound wave, also includes that facilitates the Reversible binding between antigen, antibody to react quickly reaches certainly, and this is caused Dependence test is operated can be to complete than speed faster.

As described above, the presence of the Micropump so that the miniature ultrasonic transducer units its ultrasonic emitting intensity can be allowed to fit Degree is reduced, then, the feature contributes to protecting the working electrode its sensitive coating, is allowed to from ultrasound injury.

Based on this case scheme, repair completely without the surface chemistry carried out for the polydimethylsiloxane substrate its relevant surfaces Decorations or surface chemical modification operation, therefore, this surface chemical modification layer or surface chemical modification layer not need presence, So, its body phase interior polymer molecule of the polydimethylsiloxane substrate is constantly diffused to the surface, migrated caused by its institute automatically The damaging influence of the surface chemical modification layer or surface chemical modification layer is not also just present.

The technical scheme of this case has dissolved its application related one to polydimethylsiloxane substrate addressed above totally Row technical barrier.Based on this case scheme, the polydimethyl siloxane material of this kind very cheap and easily processing and fabricating just has can Can in the micro-fluidic chip, it be prepared, production, application 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, and the structure is provided One function of facilitating the device to disassemble, thus, the hinge-type fixture is together with miniature ultrasonic transducer units appended in its hinge Just easily can be mutually disengaged with the micro-fluidic chip, then, the component that the part can freely depart from just can circulate benignly ground Reuse many times；The architectural feature is conducive to saving the use cost of the device.

The framework of this case micro-fluidic chip its large-scale integrated, determines it among actual test application, to Serum samples Requirement it is less, this body and mind for contributing to reducing related subject is damaged.

Description of the drawings

Fig. 1 is its rough outside side view of this case micro flow control chip device.

In figure, 1 is hinge-type fixture, and 2 indicate linkwork positions, and 3 is fastening screw, and 4 is higher-order of oscillation telecommunication Number transmission cable, 5 is miniature ultrasonic transducer units, and 6,11 is respectively two different hinges of locations of structures, and 7 is cover plate, 8 It is substrate, 9 is sample introduction end, and 10 is terminal, and 12 is nut, and 13 is infusion tube, and 14 is Micropump；The conjunction in legend Page clamp structure is only the legend structure illustrated, and actual its structure of the hinge-type fixture is not limited to legend hinge-type fixture knot Structure；Arrow in legend indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, its test liquid stream Flow direction.

Specific embodiment

In this case that Fig. 1 is shown embodiment, the example its main points are that the structure of the device includes micro-fluidic chip, should The structure of micro-fluidic chip includes the substrate 8 installed together bonded to each other and cover plate 7, and the substrate 8 and cover plate 7 are plate Contain the conduit formed via mould pressing process or etching technics in shape thing or tablet, that face towards the cover plate 7 of the substrate 8 Structure, substrate 8 being installed together bonded to each other has been built into the micro-fluidic chip containing pipeline configuration jointly with the cover plate 7, The locations of structures of the pipeline is located at the substrate 8 and the cover plate 7 juncture area bonded to each other, and the two ends of the pipeline are micro- with this respectively The sample introduction end 9 of fluidic chip and terminal 10 connect, and the sample introduction end 9 is the injection end of the micro-fluidic chip sample solution, should Terminal 10 is the terminal of sample solution flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal 10 and the sample introduction End 9 is located remotely from each other, the distance between the terminal 10 and the sample introduction end 9 between 3 centimetres and 10 centimetres, in the pipeline On diverse location in order or backward is equiped with working electrode and to electrode and reference electrode, the order refers to the ginseng Than its locations of structures of electrode closer to the position of the terminal 10, the backward refer to the reference electrode locations of structures closer to The position of sample introduction end 9, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumor mark The gold size sensitive membrane of will thing antibody is constituted, and the construction of the pipeline is presented parallel construction, and the pipeline in parallel construction is by 15 Lateral is in parallel to be constituted, described to be presented the profile that the pipeline of parallel construction its appearance profile is similar to parallel circuit, described The quantity of working electrode is 15, and the installation position of 15 working electrodes is located at respectively in 15 laterals, And, the tumor markerses antibody in the sensitive membrane structure of its top layer gold size of 15 working electrodes is respectively anti-to tumor markerses 15 kinds of tumor markerses antibody materials of proper energy specific binding, 15 kinds of antibody materials are respectively tumor markerses antibody AFP、CEA、CA242、CA125、CA199、CA153、CA724、CA50、NSE、CYFRA21-1、FPSA、TPSA、 β-hCG, SCCA and β 2-MG, the antigen is the antigen of broad sense, and the antibody is the antibody of broad sense, the work electricity Extremely material is argent material, gold material, carbon material or thermal decomposition conducting polymer material, the working electrode its shape Looks are presented column, lamellar or thread, and the substrate 8 its material is polydimethylsiloxane material, and its surface of substrate 8 is primary The surface of form, the surface of the primary form its be intended to refer to not change through any surface chemical modification or any surface chemistry Property the material primary form surface, the structure of the device also includes hinge-type fixture 1, the hinge-type fixture 1 its profile Profile be similar to hinge, the hinge-type fixture 1 by two hinges 6,11 being mutually hinged and through two hinges 6, 11 fastening screw 3 and one match with the fastening screw 3 and with the fastening screw 3 connect together for handss The nut 12 of dynamic fastening and hand break loose is constituted, and the nut 12 employs in this example the butterfly spiral shell for being easier manual operation Cap, butterfly nut is commercially available, and two hinges 6,11 of the hinge-type fixture 1 are respectively mutually drawn close with its tip and clamp this Micro-fluidic chip, the tip is positioned in the locations of structures of neighbouring described terminal 10 of the micro-fluidic chip, at least in A hinge on attach fixation be equiped with miniature ultrasonic transducer units, in this example, the miniature ultrasonic transducer units 5 be patch It is attached to be fixed in hinge 6, and, higher-order of oscillation electric signal transmission cable 4, the one of the higher-order of oscillation electric signal transmission cable 4 End links together with the miniature ultrasonic transducer units 5；The hinge-type fixture 1 provides a function of facilitating the device to disassemble； The miniature ultrasonic transducer units 5 its major functions is the ultrasound wave launched using it when the actual sample introduction of micro-fluidic chip is tested To reduce the interfacial tension between sample solution and the inwall of its inner passage of micro-fluidic chip so as to can be compatible, also, Using the distance between the sample introduction end 9 and the terminal 10 and the installation position of miniature ultrasonic transducer units 5 difference and Difference on its ultrasonic intensity experienced, its interfacial tension of sample introduction end 9 described in induced synthesis and its boundary of the terminal 10 Difference between the tension force of face, what the interfacial tension difference between the micro-fluidic chip two ends 9,10 can be in the micro-fluidic chip should Pressure gap is formed between two ends 9,10, the pressure gap can drive sample solution to flow to the direction of the terminal 10；This is micro- Type ultrasonic transducer 5 its function also include the ultrasound wave launched with it check biomacromolecule contained in sample its Absorption on the micro-fluidic chip its inner passage inner surface, and then check the substrate 8 of the polydimethylsiloxane material its body phase The effect of swallowing up to the biomacromolecule；It is soft and have the substrate 8 of the polydimethylsiloxane material of elasticity its function include with Its property to the strong absorption of ultrasound wave, is absorbed strongly to ultrasound wave, and thereby in the micro-fluidic chip, the terminal 10 is arrived The rapid decrement of ultrasonic intensity is realized within limited short distance between the sample introduction end 9；And, Micropump 14, the Micropump 14 are connected with the sample introduction end 9；The function of the Micropump 14 is, in the sample solution and its inner passage of the micro-fluidic chip Interfacial tension between wall by the ul-trasonic irradiation and reduce, the alternate compatibility increase precondition under, with the Micropump 14 The mechanicalness pumping strength driving force that it is brought come the interfacial tension difference between the two ends 9,10 of ultrasound wave induction Amount is supported mutually, adjusts mutually, is mutually coupled, and one is pooled in the way of Collaboration and drives test liquid to flow to the terminal The strength of 10 directions flowing.

Arrow in legend indicate the micro-fluidic chip its in actual motion, by two ends 9,10 pressure differentials drive, its The flow direction of test liquid stream.

Fig. 1 is depicted without the associate members such as the higher-order of oscillation electric signal generator.

The Micropump 14 can be customized to specialized factory.

Described its main body of hinge-type fixture can simply be changed a social system using commercially available hinge；Can also customize to five metals processing factory.

Involved miniature ultrasonic transducer units 5 are commercially available；Can also customize to ultrasonic transducer producer.

Involved higher-order of oscillation electric signal transmission cable 4 is commercially available；Can also be to ultrasonic transducer producer or cable special manufacturer Family's customization.

Involved higher-order of oscillation electric signal generator market has the product for being close to needs commercially available；Can also customize to relevant manufacturers.

Its inner passage of the micro-fluidic chip involved by this case is the pipeline of hydrophobic capillary tube form.

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.Micropump in legend is external form Micropump；The Micropump can certainly do into or say the embedded microfluidic chip structure inside sample introduction end or its Near-neighbor Structure position The Micropump of the built in version, its basic structure key element is identical with the Micropump of external form.

The multiple tracks electrochemical workstation is commercially available；The multiple tracks electrochemical workstation can also be special to correlation according to specific needs Industry producer customizes.

In view of its form of this case above related text expresses that it is described pipeline of the presentation parallel construction is clear enough It is clear, the concrete form of the pipeline in this kind of micro-fluidic chip of this case is no longer specifically illustrating in this case embodiment.

Antibody described in this case refers to the antibody of broad sense；Antigen described in this case refers to the antigen of broad sense；Immunity is multiple described in this case Compound refers to the immune complex of broad sense.

Claims (10)

1. the common tumor markers examination chip apparatus of cheap hydrophobic substrate are related to, it is characterised in that the structure bag of the device Micro-fluidic chip is included, the structure of the micro-fluidic chip includes the substrate installed together bonded to each other and cover plate, the substrate and lid Piece is plate object or tablet, and what is formed via mould pressing process or etching technics contained in that face towards the cover plate of the substrate Channel structure, substrate being installed together bonded to each other has been built into the micro-fluidic core containing pipeline configuration jointly with the cover plate Piece, the locations of structures of the pipeline is located at the substrate and cover plate juncture area bonded to each other, and the two ends of the pipeline are micro- with this respectively The sample introduction end of fluidic chip and terminal connect, and the sample introduction end is the injection end of the micro-fluidic chip sample solution, and the terminal is this The terminal that sample solution flows in its chip when the actual sample introduction of micro-fluidic chip is tested, the terminal is located remotely from each other with the sample introduction end, should The distance between terminal and the sample introduction end are in order or inverse on diverse location in the pipeline between 3 centimetres and 10 centimetres Sequence is equiped with working electrode and to electrode and reference electrode, and the order refers to the reference electrode its locations of structures and more leans on The nearly terminal location, the backward refers to the reference electrode locations of structures closer to the sample introduction end position, the work Electrode is made up of conductive electrode and the gold size sensitive membrane for having embedded tumor markerses antibody being attached on the conductive electrode, The construction of the pipeline is presented parallel construction, and the pipeline in parallel construction is made up of 15 lateral parallel connections, the presentation The pipeline of parallel construction its appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is 15, should The installation position of 15 working electrodes is located in 15 laterals respectively, and, 15 working electrode its tables Tumor markerses antibody in layer gold size sensitivity membrane structure is respectively that 15 kinds can specifically bound to tumor markerses antigen swell Tumor markers antibody materials, 15 kinds of antibody materials be respectively tumor markerses antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, it is described anti- Original is the antigen of broad sense, and the antibody is the antibody of broad sense, the working electrode its material be argent material, gold material, Carbon material or thermal decomposition conducting polymer material, the working electrode its pattern is presented column, lamellar or thread, the substrate its Material is polydimethylsiloxane material, and its surface of the substrate is the surface of primary form, the surface of the primary form its be meant to Be not through any surface chemical modification or the surface of the primary form of the material of any surface chemical modification, the device Structure also includes hinge-type fixture, the hinge-type fixture its appearance profile likeness in form hinge, and the hinge-type fixture is by being hinged one Two hinges and a fastening screw through two hinges and one for rising match with the fastening screw and with the fastening The nut for fastening and hand break loose manually that screw connects together is constituted, and two hinges of the hinge-type fixture are respectively with its end The micro-fluidic chip is mutually drawn close and clamped to the tip, and the tip is positioned at the locations of structures of the neighbouring described terminal of the micro-fluidic chip On, fixation is attached in a hinge at least in and is equiped with miniature ultrasonic transducer units, and, higher-order of oscillation telecommunication Number transmission cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable link together；The hinge Formula fixture provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units its major functions is in micro-fluidic chip When actual sample introduction is tested, the ultrasound wave launched using it is reducing the inwall of sample solution and its inner passage of micro-fluidic chip Between interfacial tension so as to can be compatible, also, using the sample introduction end and the terminal and the miniature ultrasonic transducing Difference in the distance between device installation position difference and its ultrasonic intensity for being experienced, sample introduction end described in induced synthesis its Difference between interfacial tension and the terminal its interfacial tension, the interfacial tension difference between the micro-fluidic chip two ends can be Pressure gap is formed between the two ends of the micro-fluidic chip, the pressure gap can drive sample solution to the end flow；Should Miniature ultrasonic transducer units its functions also include the ultrasound wave launched with it check biomacromolecule contained in sample its Absorption on the micro-fluidic chip its inner passage inner surface, and then check the substrate of the polydimethylsiloxane material its body phase pair The effect of swallowing up of the biomacromolecule；It is soft and have the substrate of the polydimethylsiloxane material of elasticity its function include it is right with its The property that ultrasound wave absorbs strongly, is absorbed strongly to ultrasound wave, and thereby in the micro-fluidic chip terminal to the sample introduction end Between limited short distance within realize the rapid decrement of ultrasonic intensity；And, Micropump, the Micropump is connected with the sample introduction end； The function of the Micropump is that the interfacial tension between the inwall of the sample solution and its inner passage of micro-fluidic chip receives the ultrasound Ripple acts on and reduces, under the precondition that the alternate compatibility increases, pumps strength to lure with the ultrasound wave with the mechanicalness of the Micropump Interfacial tension difference between the two ends led its driving force brought is supported mutually, adjusts mutually, is mutually coupled, with The mode of Collaboration pools one and drives test liquid to flow to the strength of the terminal direction flowing.

2. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the pipeline is capillary channel including the lateral.

3. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the thermal decomposition conducting polymer is the conductive material formed Jing after anoxybiotic heat treatment by polyimides or polyacrylonitrile.

4. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature Be, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the working electrode Length is between 1 micron to 15000 microns.

5. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that its material of the cover plate in structure is polydimethylsiloxane material.

7. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the Micropump is miniature piezoelectric pump, miniature peristaltic pump or miniature air driven pump.

8. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the cover plate in structure and substrate its thickness are between 1.0 millimeters and 5.0 millimeters.

9. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature Be that the structure of the micro fluidic device also includes higher-order of oscillation electric signal generator, the higher-order of oscillation electric signal transmission cable its The other end is connected with the higher-order of oscillation electric signal generator.

10. common tumor markers examination chip apparatus for being related to cheap hydrophobic substrate according to claim 1, its feature It is that the miniature ultrasonic transducer units its specified ultrasonic emitting power is between 5 milliwatts and 5000 milliwatts, and this is miniature super The frequency of acoustic wave transducer its ultrasound wave operationally launched is between 100KHz and 12MHz.