CN107335486A - The classifiable tumor markers in detecting micro flow control chip device that male is applicable - Google Patents

Abstract

The present invention relates to the classifiable tumor markers in detecting micro flow control chip device that a kind of male is applicable, belong to analysis testing field.The substrate of male's classifiable tumor markers in detecting micro-fluidic chip is made with dimethyl silicone polymer is PDMS, there are many advantages, but there is also serial problem；There is the PDMS spontaneous slowly deformation to be inclined to and thus cause its pipeline of chip to become narrow gradually, and PDMS surfaces are strongly hydrophobic, and surface chemical modification modified effect is difficult to persistently, its surface and can also adsorb, swallow up large biological molecule.This case main points are that substrate material is the PDMS for having ecosystem surface, and micron silica particle is filled with its pipeline of chip；Its function of the micron silica granule filler includes supporting pipeline inwall, and, with the set of hydrophily fluid channel come the hydrophobic pipeline of compensatory script, and, by hard squeeze, cover the chance for thereby reducing large biological molecule and being contacted with the internal face in inner-walls of duct face.

Description

The classifiable tumor markers in detecting micro flow control chip device that male is applicable

Technical field

The present invention relates to the classifiable tumor markers in detecting micro flow control chip device that a kind of male is applicable, and belongs to analysis test Field.

Background technology

Tumor markers (tumor marker, TM) refers in the generation and breeding of tumour, is produced in itself by tumour cell Raw is either reacted and caused, a kind of material that reflection tumour exists and grown tumour cell by body, including protein, Hormone, enzyme (isodynamic enzyme) and oncoprotein etc..The tumor markers in blood samples of patients or body fluid is chemically examined, can be in cancer screening Early detection tumour, and the effect of observe oncotherapy and judge patient's prognosis.The tumor markers clinically commonly used at present has： (1) alpha-fetoprotein (AFP) is the mark of the tumours such as primary carcinoma of liver, carcinoma of testis, oophoroma；(2) carcinomebryonic antigen (CEA) is digestion The mark of the tumours such as system tumor, lung cancer, breast cancer；(3) CA125 (CA125) is the mark of the tumours such as oophoroma； (4) CA153 (CA153) is the mark of the tumours such as breast cancer；(5) CA19-9 (CA19-9) is digestive system tumor Mark；(6) CA724 (CA724) is the mark of the tumours such as stomach cancer, oophoroma；(7) carbohydrate antigen 242 (CA242) For the mark of digestive system tumor；(8) CA50 (CA50) is the mark of the tumours such as digestive system tumor, breast cancer, lung cancer Thing；(9) CYFRA21-1 (Cy211) is the mark of the tumours such as non-small cell lung cancer；(10) neuronspecific enolase (NSE) For the mark of the tumours such as ED-SCLC, neuroendocrine tumor；(11) PSA (PSA) is prostate cancer Tumor markers；(12) human chorionic gonadotrophin (HCG) is that embryonic cell carcinoma, trophoblastic tumor (suede cancer, vesicular mole) etc. are swollen The mark of knurl；(13) thyroglobulin (TG) is the mark of thyroid cancer；(14) ferritin (SF) is digestive system tumor, liver The mark of the tumours such as cancer, breast cancer, lung cancer；(15) B2M (β 2-MG) is in chronic lymphocytic leukemia, lymph Raised in the patient body fluids such as knurl, myeloma, lung cancer, thyroid cancer, nasopharyngeal carcinoma；(16) squamous cell antigen (SCC) be cervical carcinoma, The tumor markerses such as lung squamous cancer, the cancer of the esophagus.The tumor markers overwhelming majority clinically detected at present is not only present in malignant tumour In, exist in benign tumour, embryonic tissue, even in normal structure.Therefore, tumor markers has dynamic chek and multinomial Joint inspection is more valuable.So for numerous tumor markerses, clinically how to selectDifferent tumours understands some phases Breast cancer is often appeared in special tumor markers, such as CA153；CEA often appears in intestinal cancer, stomach cancer；CA19-9 is often appeared in Intestinal cancer, cancer of pancreas；CA125 often appears in oophoroma etc..Clinician can be according to the different mark of different tumor examinations. Same tumour or different types of tumour can have one or more of tumor markerses abnormal；Same tumor markers can be in difference Tumour in occur.To improve the additive diagnostic value of tumor markers and determining which kind of mark can be as the follow-up prison after treatment Index is surveyed, tumor markers joint-detection, the complementary tumor markers 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 markers 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 may refer to soon Before the monograph " diagram Microfluid based Lab on a chip " that goes out, the monograph is published via Science Press, and the monograph is for micro-fluidic Past of technology, now, and, vision of the future etc. etc., suffer from detail, be deep into long discussion of detail.

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

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

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

But thing is really not so simple.

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

According to the literature, there is a kind of expedient solution, be that a small amount of surfactant is added in sample to be tested solution, with Just during test, dynamic, temporary transient effective surface hydrophilic layer is built temporarily in the PDMS micro-channels inner surface；So And due to the amphipathic characteristic of surfactant, the introduced surfactant of the program it is inevitable simultaneously also can with sample solution Combination occurs for tested composition so that tested composition can not be detected normally or it is detected ratio and dubiously reduced；The table Face activating agent is possibly even fully wrapped around by tested composition with micelle form so that tested composition can not be detected or identify；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 inner surface can be released Decorating state can not persistently, chip can not largely make, largely lay in and then be widely popularized and such a make the numerous specialties in this area The puzzlement that personnel are entangled with for a long time, the highly difficult problem that exactly one its obvious technology barrier can not despise.

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

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

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

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

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

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

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

The content of the invention

The technical problem to be solved by the invention is to provide a package solution, solve what is addressed above totally The problem of three aspects, also, the solution is applied to build a kind of new can be directed to and belongs to male's physical examination and especially needs To be paid close attention to seven kinds carry out the micro flow control chip device of examination simultaneously while detection than more typical primary tumor mark.

The present invention solves the technical problem by following scheme, and the device that the program provides is that the typical case that a kind of male is applicable swells Tumor markers joint-detection micro flow control chip device, program feature are that the structure of the device includes multichannel micro-fluidic chip, The structure of the micro-fluidic chip includes being bonded to each other installing substrate and cover plate together, the substrate and cover plate be plate object or The channel structure via mould pressing process or etching technics formation, the base are contained in tablet, that face towards the cover plate of the substrate Piece also contains and is connected and pierces being formed via mould pressing process, etching technics or simple drilling technology for the substrate with the channel structure Window structure, be bonded to each other the substrate being installed together and be built into jointly containing pipeline configuration and phase therewith with the cover plate The micro-fluidic chip of liquid pool structure even, the locations of structures of the pipeline are located at the interface zone that the substrate is bonded to each other with the cover plate, 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 locations of structures of the liquid pool, institute Stating liquid pool has two kinds, and two kinds of liquid pools are the sample introduction end liquid pool and terminal liquid pool positioned at different structure position respectively, and this is micro-fluidic The sample introduction end position of chip has one or more sample introduction end liquid pool, and the sample introduction end refers to micro-fluidic chip reality The injection end position of detected solution during the sample introduction of border, then there is a terminal liquid pool in the terminal location of the micro-fluidic chip, it is described Terminal refers to the terminal location of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal and the sample introduction end It is located remotely from each other, one end of the pipeline and the sample introduction end liquid pool UNICOM positioned at sample introduction end, the other end of the pipeline is micro- with being located at this The terminal liquid pool UNICOM of the terminal of fluidic chip, and, sequentially or backward be respectively installed in the pipeline it is different Working electrode on position and to electrode and reference electrode, the order refers to that its locations of structures of the reference electrode is more leaned on The nearly terminal location, the backward refer 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 markers antibody being attached on the conductive electrode, Parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is made up of seven lateral parallel connections, and the presentation is simultaneously Its appearance profile of the pipeline of connection construction is similar to the profile of parallel circuit, and the quantity of the working electrode is seven, this seven The installation position of working electrode is located in seven laterals respectively, and, its top layer gold size of seven working electrodes is sensitive Tumor markers antibody in membrane structure is the seven kinds of tumor markers antibody that can be specifically bound to tumor markers antigen respectively Material, seven kinds of antibody materials are tumor markers antibody CA199, CA242, AFP, EB, CEA, TPSA and FPSA respectively, The antigen is the antigen of broad sense, and the antibody is the antibody of broad sense, and its material of the working electrode is argent material, gold Column, sheet or thread is presented in material, carbon material or thermal decomposition conducting polymer material, its pattern of the working electrode, with And Micropump, the Micropump are connected with the sample introduction end of the micro-fluidic chip, the Micropump is the Micropump or built in version of external form Micropump, its material of the substrate are dimethyl silicone polymer materials, and its surface of the substrate is the surface of primary form, the primary form Surface its be intended to refer to the primary form of the not material by any surface chemical modification or any surface chemical modification Surface；And many silica dioxide granules, many silica dioxide granules are filled in the pipeline, the pipeline Its tube chamber is filled up by many silica dioxide granules, and its particle size range of the silica dioxide granule is micro- with 200 between 10 microns Between rice, the silica dioxide granule is silica crystalline particles, silica glass granule or silica composition its shared weight Measure inorganic glass particles of the percentage more than 80%.

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

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

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

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

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

The silica dioxide granule can also customize to specialized factory.

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

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

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

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

Involved Micropump can customize to specialized factory.

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

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

The tumor markers antibody is the antibody of broad sense, and the antibody of the broad sense refers to possessing antibody function or functionally similar In antibody can occur with tumor markers involved by various corresponding clinics combine and formed immune complex or immune compound The material of the analog of thing；The tumor markers antigen is the antigen of broad sense, and the antigen of the broad sense refers to utilize accordingly 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.

The gold size sensitive membrane is to be sufficiently mixed chitosan gold size solution and tumor markers antibody-solutions uniformly, uses point sample instrument Point sample is coated on specified structure position, and forms its drying and forming-film.Tumor markers antibody in the gold size sensitive membrane is equal The tumor markers antibody marked for horseradish peroxidase or glucose oxidase, the gold size sensitive membrane have been included as fixing Above-mentioned each tumor markers antibody and introduce complementary medium therein, the complementary medium for example chitosan, 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, still, for prepare liquid sample and the consideration of reagent loss etc. is reduced less as far as possible, the pipeline includes described The passage of the preferred capillary level of lateral, the passage of the capillary level imply that the interior of its internal diameter and the capillary on ordinary meaning The suitable passage in footpath.The shape of cross section of its inner passage of capillary can be arbitrary shape, the shape of cross section example Such as circular, oval, square, rectangle, bar shaped, naturally it is also possible to it is the linear of bending arbitrarily be present, 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 the electrode of microsize to electrode and reference electrode, and its electrode shape may each be any choosing Fixed shape, the arbitrarily selected shape such as column, sheet, strip or thread etc..It is described to electrode and the ginseng Art-recognized meanings than the electrode vocabulary of itself are known.

It is related to several liquid pools in this case microfluidic chip structure, the liquid pool is the pond shape or scrotiform structure for transitional liquid storage Make, its shape of the inner chamber of each liquid pool may each be arbitrarily selected shape, the cavity shape such as cylindrical empty Cavity-like, square column type cavity-like, oblong cavity shape or spherical hollow space shape etc..

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

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

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

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

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

The terminal and the distance between the sample introduction end are unlimited；But the distance between the terminal and described sample introduction end Preferred scope is between 3 centimetres and 10 centimetres.

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

The application method of this case micro-fluidic chip：

This case is flowed with Micropump driving liquid stream in the hydrophobic capillary channel of seven channel microfluidic chip, utilizes multichannel Electrochemical analytical instrument carries out joint-detection to seven kinds of tumor markers antigens respectively.

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

1st, blood serum sample liquid is added in micro-pipe road, under Micropump driving, various tumor markers antigen molecules are by each logical In road on electrode surface gold size sensitive membrane embed corresponding horseradish peroxidase-labeled tumor markers antibody capture.

2nd, the tumor markers antibody of horseradish peroxidase-labeled is formed immune multiple with the tumor markers antigen in blood serum sample Compound.

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

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

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

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

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

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

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

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

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

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

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

Its highly integrated structure of this case micro-fluidic chip, determines it among practical application to the requirement of test serum Smaller, this contributes to the body and mind damage for reducing related subject.

Brief description of the drawings

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

In figure, 1 is the substrate of dimethyl silicone polymer material, and 2 be cover plate.

Embodiment

In this case that Fig. 1 is shown embodiment, the example is characterized in, the structure of the device includes multichannel micro-fluidic core Piece, the structure of the micro-fluidic chip include being bonded to each other the substrate 1 and cover plate 2 of installing together, the substrate 1 and cover plate 2 It is plate object or tablet, that face towards the cover plate 2 of the substrate 1 is contained to be formed via mould pressing process or etching technics Channel structure, the substrate 1 also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics Or the window structure that simple drilling technology is formed, it is bonded to each other the substrate 1 being installed together and is built into jointly with the cover plate 2 Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline are located at the substrate 1 with being somebody's turn to do The interface zone that cover plate 2 is bonded to each other, its side of the window is blocked by the cover plate 2 and opposite side opens, the structure bit of the window Put be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are the sample introduction positioned at different structure position respectively End liquid pool and terminal liquid pool, the sample introduction end position of the micro-fluidic chip have one or more sample introduction end liquid pool, institute The injection end position that sample introduction end refers to detected solution during the micro-fluidic chip actual sample introduction is stated, in the terminal position of the micro-fluidic chip Put, there is a terminal liquid pool, the terminal refers to liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested Terminal location, the terminal is located remotely from each other with the sample introduction end, and one end of the pipeline joins with the sample introduction end liquid pool positioned at sample introduction end It is logical, the other end of the pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip, and, sequentially or The working electrode that is respectively installed in backward in the pipeline on diverse location and to electrode and reference electrode, what the order referred to Be its locations of structures of the reference electrode closer to the terminal location, the backward refers to the reference electrode locations of structures more Close to the sample introduction end position, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumour The gold size sensitive membrane of mark antibody is formed, and parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is by seven Lateral is in parallel to be formed, and its appearance profile of the pipeline of the presentation parallel construction is similar to the profile of parallel circuit, described The quantity of working electrode is seven, and the installation position of seven working electrodes is located in seven laterals respectively, and, Tumor markers antibody in its top layer gold size sensitivity membrane structure of seven working electrodes is respectively can be special to tumor markers antigen The opposite sex combine seven kinds of tumor markers antibody materials, seven kinds of antibody materials be respectively tumor markers antibody CA199, CA242, AFP, EB, CEA, TPSA and FPSA, the antigen are the antigen of broad sense, and the antibody is the antibody of broad sense, the work It is argent material, gold material, carbon material or thermal decomposition conducting polymer material to make electrode its material, the working electrode Column, sheet or thread is presented in its pattern, and, Micropump, the Micropump is connected with the sample introduction end of the micro-fluidic chip, described micro- Pump is the Micropump of external form or the Micropump of built in version, and its material of the substrate 1 is dimethyl silicone polymer material, the substrate 1 Its surface is the surface of primary form, the surface of the primary form its be intended to refer to not by any surface chemical modification or appoint The surface of the primary form of the material of what surface chemical modification；And many silica dioxide granules, many titanium dioxides Silicon grain is filled in the pipeline, and its tube chamber of the pipeline is filled up by many silica dioxide granules, the silica For its particle size range of grain between 10 microns and 200 microns, the silica dioxide granule is silica crystalline particles, titanium dioxide Silica glass particle or silica composition its inorganic glass particles of shared percentage by weight more than 80%.

Fig. 1 is depicted without the Micropump.

Also the associate members such as multiple tracks electrochemical workstation are depicted without in Fig. 1.

The multi-channel electrochemical work station can be with commercially available；The multi-channel electrochemical work station can also be according to specific Whereabouts specialised manufacturers are needed to customize.

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

In view of the pipeline of the presentation parallel construction its form that this case related text above expresses that it is described 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；It is immunized described in this case multiple Compound refers to the immune complex of broad sense.

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

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

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

Involved Micropump can customize to specialized factory.

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

Claims (9)

1. the classifiable tumor markers in detecting micro flow control chip device that male is applicable, it is characterised in that the structure of the device Including multichannel micro-fluidic chip, the structure of the micro-fluidic chip includes being bonded to each other the substrate and cover plate of installing together, described Substrate and cover plate are plate object or tablet, and that face towards the cover plate of the substrate is contained via mould pressing process or etching work Skill formed channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etch The window structure that technique or simple drilling technology are formed, it is bonded to each other the substrate being installed together and is built into jointly with the cover plate Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline are located at the substrate and the lid The interface zone that piece is bonded to each other, 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 locations of structures of the liquid pool, the liquid pool have two kinds, and two kinds of liquid pools are the sample introduction end liquid pool positioned at different structure position respectively And terminal liquid pool, the sample introduction end position of the micro-fluidic chip have one or more sample introduction end liquid pool, the sample introduction End refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, then has in the terminal location of the micro-fluidic chip One terminal liquid pool, the terminal refer to the terminal of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested Position, the terminal are located remotely from each other with the sample introduction end, one end of the pipeline and the sample introduction end liquid pool UNICOM positioned at sample introduction end, should The other end of pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip, and, sequentially or backward The working electrode that is respectively installed in the pipeline on diverse location and to electrode and reference electrode, the order refers to described For its locations of structures of reference electrode closer to the terminal location, the backward refers to the reference electrode locations of structures closer to institute State sample introduction end position, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumor markers The gold size sensitive membrane of antibody is formed, and parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is by seven branched pipes Road is in parallel to be formed, and its appearance profile of the pipeline of the presentation parallel construction is similar to the profile of parallel circuit, the work electricity The quantity of pole is seven, and the installation position of seven working electrodes is located in seven laterals respectively, and, this seven Tumor markers antibody in its top layer gold size sensitivity membrane structure of working electrode is respectively can specificity knot to tumor markers antigen Close seven kinds of tumor markers antibody materials, seven kinds of antibody materials be respectively tumor markers antibody CA199, CA242, AFP, EB, CEA, TPSA and FPSA, the antigen are the antigen of broad sense, and the antibody is the antibody of broad sense, the working electrode Its material is argent material, gold material, carbon material or thermally decomposes conducting polymer material, its pattern of the working electrode Column, sheet or thread is presented, and, Micropump, the Micropump is connected with the sample introduction end of the micro-fluidic chip, and the Micropump is outer The Micropump of form or the Micropump of built in version are put, its material of the substrate is dimethyl silicone polymer material, and its surface of the substrate is former The surface of raw form, the surface of the primary form its be intended to refer to and do not pass through any surface chemical modification or any surface chemistry The surface of the primary form of the modified material；And many silica dioxide granules, many silica dioxide granule fillings In the pipeline, its tube chamber of the pipeline is filled up by many silica dioxide granules, its particle diameter model of the silica dioxide granule Enclose between 10 microns and 200 microns, the silica dioxide granule is silica crystalline particles, silica glass granule Or silica composition its inorganic glass particles of shared percentage by weight more than 80%.

2. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is that it is capillary channel that the pipeline, which includes the lateral,.

3. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is that the thermal decomposition conducting polymer is the electric conductivity material formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated Material.

4. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the working electrode Length between 1 micron to 15000 microns.

5. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is that the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is, the cover plate its material in structure is dimethyl silicone polymer material.

7. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is, the distance between the terminal and the sample introduction end between 3 centimetres and 10 centimetres, the cover plate and substrate its Thickness is between 1.0 millimeters and 5.0 millimeters.

8. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is that its particle size range of the silica dioxide granule is between 100 microns and 200 microns.

9. the classifiable tumor markers in detecting micro flow control chip device that male according to claim 1 is applicable, it is special Sign is that its inside diameter ranges of the pipeline are between 500 microns and 1000 microns.