CN107694638A

CN107694638A - The tumor markers joint-detection six-channel microfluidic chip device simplified

Info

Publication number: CN107694638A
Application number: CN201610677021.6A
Authority: CN
Inventors: 宋岳
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-08-09
Filing date: 2016-08-09
Publication date: 2018-02-16

Abstract

The present invention relates to a kind of tumor markers joint-detection six-channel microfluidic chip device simplified, belong to analysis testing field.Dimethyl silicone polymer be PDMS its be used for making the substrate of the multichannel combined detection micro-fluidic chip of classifiable tumor mark, it is advantageous, while a series of problems for needing to overcome be present；This case is directed to the serial problem.This case main points are, the selected PDMS with ecosystem surface of substrate, and the chain ring type magnetic holding device for being equiped with miniature ultrasonic transducer units socket is positioned at the sample liquid stream terminal of the micro-fluidic chip its neighbor positions, interfacial tension is reduced with ultrasonic wave, the interface compatibility between related solid-liquid two-phase is significantly increased, strong absorbability using PDMS to ultrasonic wave simultaneously, reach ultrasonic intensity rapid decrement in short distance, so as to form interfacial tension difference at the both ends of the chip, sample liquid stream is thereby facilitated to be flowed along originally hydrophobic capillary channel to terminal direction.The device exempts to use Micropump completely.

Description

The tumor markers joint-detection six-channel microfluidic chip device simplified

Technical field

The present invention relates to a kind of tumor markers joint-detection six-channel microfluidic chip device simplified, belong to analysis Testing field.

Background technology

Tumor markers (tumor marker, TM) refers in the generation and breeding of tumour, by tumour cell sheet Either tumour cell is reacted and caused, a kind of material of the presence of reflection tumour and growth by body caused by body, bag Include protein, hormone, enzyme (isodynamic enzyme) and oncoprotein etc..The tumor markers in blood samples of patients or body fluid is chemically examined, can be Early detection tumour in cancer screening, and the effect of observe oncotherapy and judge patient's prognosis.Clinically commonly use at present Tumor markers has：(1) alpha-fetoprotein (AFP) is the mark of the tumours such as primary carcinoma of liver, carcinoma of testis, oophoroma；(2) cancer embryo Antigen (CEA) is the mark of the tumours such as digestive system tumor, lung cancer, breast cancer；(3) CA125 (CA125) is ovary The mark of the tumours such as cancer；(4) CA153 (CA153) is the mark of the tumours such as breast cancer；(5) CA19-9 (CA19-9) it is the mark of digestive system tumor；(6) CA724 (CA724) is the mark of the tumours such as stomach cancer, oophoroma Thing；(7) carbohydrate antigen 242 (CA242) is the mark of digestive system tumor；(8) CA50 (CA50) is digestive system The mark of the tumours such as tumour, breast cancer, lung cancer；(9) CYFRA21-1 (Cy211) is the mark of the tumours such as non-small cell lung cancer； (10) neuronspecific enolase (NSE) is the mark of the tumours such as ED-SCLC, neuroendocrine tumor；(11) before Row gland specific antigen (PSA) is the tumor markers of prostate cancer；(12) human chorionic gonadotrophin (HCG) is that embryo is thin The mark of the tumours such as born of the same parents' cancer, trophoblastic tumor (suede cancer, vesicular mole)；(13) thyroglobulin (TG) is the mark of thyroid cancer Will thing；(14) ferritin (SF) is the mark of the tumours such as digestive system tumor, liver cancer, breast cancer, lung cancer；(15) β 2- microballoons Albumen (β 2-MG) is in patient body fluids such as chronic lymphocytic leukemia, lymthoma, myeloma, lung cancer, thyroid cancer, nasopharyngeal carcinoma Middle rise；(16) squamous cell antigen (SCC) is the tumor markerses such as cervical carcinoma, lung squamous cancer, the cancer of the esophagus.Clinically detect at present The tumor markers overwhelming majority be not only present in malignant tumour, exist in benign tumour, even embryonic tissue, normal group In knitting.Therefore, tumor markers has dynamic chek and multinomial joint inspection more valuable.So for numerous tumor-markers Thing, clinically how to selectDifferent tumours understands some relatively special tumor markerses, as CA153 often appears in mammary gland Cancer；CEA often appears in intestinal cancer, stomach cancer；CA19-9 often appears in intestinal cancer, cancer of pancreas；CA125 often appears in oophoroma etc..Face Bed doctor can be according to the different mark of different tumor examinations.Same tumour or different types of tumour can have a kind of or several Kind tumor markers is abnormal；Same tumor markers can occur in different tumours.To improve the auxiliary of tumor markers Which kind of mark can carry out tumor markers joint-detection to diagnostic value as the follow-up visit monitoring index after treatment with determination, The complementary tumor markers composition best of breed of several sensitivity of reasonable selection, specific performance, carries out joint-detection.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, can be with Referring to following Chinese invention patent application case：CN200410041175.3、CN200510026780.8、 CN200610040051.2、CN200910064647.X。

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

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 be etched with the substrate of small fluid course and fit together therewith Cover plate, the small fluid course on the substrate, before upper cover plate is assembled, it is apparent on see to be exactly some micro-channels, to wait until After covering cover plate thereon, just really closure forms the small fluid course, the conduit inner surface of the micro-channel together with The part cover plate that surround the micro-channel forms described small fluid course together；So, it is clear that after assembling completes The small fluid course, the major part of its inner surface area is the inner surface area of that micro-channel, in other words, should The state or property of micro-channel inner surface substantially determine the integrality or property of the small fluid course；Therefore say, this The inner surface state or inner surface property of micro-channel of the individual structure on substrate are key factors；In principle, it is any can The material of its solid forms is kept or kept substantially, can be used to make substrate and cover plate, such as, it can act as substrate and lid The material of piece can be monocrystalline silicon piece, quartz plate, sheet glass, high polymer for example dimethyl silicone polymer, polymethyl methacrylate, Makrolon etc.；Certainly, the selection of substrate and the selection of cover plate be able to can also be differed with identical；From material consumption, make From the point of view of difficulty and application popularization prospect etc. etc., not small difference, the especially choosing of that substrate between these materials be present Material, have a great influence.

In various substrate making materials, dimethyl silicone polymer, i.e. PDMS, comparatively very easily shaping, at this It is extremely simple that micro-channel is made on the substrate of sample, and the lower cost for material, base is made with the polydimethyl siloxane material Piece, micro-channel, and the cover plate phase made with the cheap material such as glass or polypropylene or other plastic sheets are being suppressed or etched thereon Coordinate, be a kind of more satisfactory selection seemingly；Certainly, patch material can also select to use cheap dimethyl silicone polymer Material：So, this substrate selection is the scheme of polydimethyl siloxane material, and material is extremely cheap, and making is extremely simple, seems It should be extremely easy to popularize, 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 Strong hydrophobic material, micro-channel is built on this material, if without being operated for the modified of the micro-channel surface, that , after overall assembling is completed, that is, after covering cover plate, because the micro-channel its inner surface in structure occupies most liquid The inner surface of circulation road, then, its strong hydrophobic property of the PDMS micro-channels inner surface, is deciding factor, it can cause Similar to the aqueous solution the fine liquid stream of polar liquid by becoming very difficult, its flow resistance is big, or even in general is micro- Pump is all 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 behaviour Make；So, this is pretty troublesome for the modified operation of PDMS micro-channel inner surfacesThat falls nor this problem, is formed tight Weight technology puzzlement, be another problem：PDMS polymer molecules inside its body phase of this PDMS material substrate have automatic The characteristic diffuse to the surface, migrated, the spy that this substrate body phase inside PDMS polymer molecules are diffused to the surface, migrated automatically Property, the state after modification by its inner surface of that micro-channel of surface modifying and decorating will be caused can not to maintain long enough Time, the holding time for micro-channel its inner surface state after that is surface-modified be substantially only sufficient to complete laboratory internal The time of test experiments needs；In other words, by surface modification or the PDMS micro-channel inner surfaces of surface modification, it is modified The surface state that is formed can not be lasting afterwards or after saying modification, but soon automatically tends to or say and become surface modification again Surface state before, the strong hydrophobic surface state of that script is returned in the shorter time, then, just think, this The micro-fluidic chip of sample can largely make, mass storage, be widely popularized, and answer is it is obvious that is, impossible.This Micro-channel on PDMS material, if not doing surface modification, similar to the aqueous solution the fine liquid stream of polar solvent can not pump it is logical Cross, chip also cannot just use；And if having done surface modification, its state after modifying can not be persistently kept again, or together Sample can not popularization and application.

So, how to accomplish that substrate can either be made using cheap PDMS material, and the microflute can be released Road inner surface decorating state can not persistently, chip can not largely make, largely lay in and then be widely popularized and such a make ability The puzzlement that the numerous professionals in domain 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 have 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, until It is heavy to be trapped within the body phase of PDMS substrates, in fact, this process, partly it is also due to PDMS material body phase interior 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. continue not Disconnectedly from inside PDMS body phases to those polymer molecules of its diffusion into the surface, migration, its result moved, be little by little by that It has been involved in a bit by the large biological molecule of adsorption within the body phase of PDMS substrates, briefly, these adsorbed biologies Macromolecular is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up the phenomenon of large biological molecule, its institute Caused by influence, necessarily cause the severe deviations of all kinds of test data of experiment for being related to large biological molecule.

As described above, the problem of PDMS substrates, is, its not only adsorption large biological molecule, and swallow up biological big point Son, so, as the large biological molecule of experiment test object, its disappearance will not stop because surface saturation is adsorbed, and It is, it is constantly adsorbed, also constantly swallowed up.

On PDMS substrates, its body phase is constantly swallowed up and tests showing for associated biomolecule macromolecular in related experiment test process As, it is to say that another kind, which is explained, substantial amounts of Minute pores in PDMS body phases be present, associated biomolecule macromolecular by after adsorption, Depression enters these Minute pores, and then is swallowed up；However, inventor thinks, those can allow the sky of miniature scale Qi leel squeezes into the Minute pores therebetween, not equal to saying that they also can directly allow the large biological molecule of relative large scale to enter Enter, both difference on yardstick are huge, must not make sweeping generalizations.Explanation is bypassed, in any case, as dependence test analysis object Large biological molecule is adsorbed by PDMS substrate micro-channels inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is person in charge of reception at ceremonies Phenomenon existing for sight.

, can be from containment PDMS surfaces pair in order to prevent swallow up effect of this PDMS substrate bodies relative to large biological molecule The absorption of large biological molecule 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 micro-channel part, by changing The micro-channel inner surface of modification is learned, its absorption to large biological molecule can be contained, and then avoid large biological molecule Swallowed up by PDMS substrate body phases；But or that old problem, that is, the chemical modification on PDMS material surface is modified Surface state afterwards can not persistently be kept, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface, move automatically The process of shifting, it soon can become that micro-channel inner surface state being modified by surface chemical modification again script strong and dredge Water 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 micro-channel 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 is enough Reach and contain the absorption process of the PDMS substrate micro-channel inner surfaces to large biological molecule for a long time, and then prevent PDMS substrate body phases The effect of swallowing up to large biological molecule so that related chip manufactured goods be able to maintain that one it is prolonged enough, reasonably guarantee the quality Phase, it is exactly a very intractable problem.The problem equally makes this area numerous as another problem addressed above Professional is entangled with, perplexed for a long time, and the problem is equally the highly difficult problem that its obvious technology barrier can not despise. Also be present many year in the problem, so far, also not yet properly settled.

The content of the invention

The technical problem to be solved by the invention is to provide a package solution, solves totally above Two problems addressed, also, the solution is applied to build a kind of new can be directed to six kinds than more typical master Tumor markers is wanted to carry out the micro flow control chip device of examination simultaneously while detection.

The present invention solves the technical problem by following scheme, and the device that the program provides is a kind of tumour mark simplified Will thing joint-detection six-channel microfluidic chip device, the program are characterized in that the structure of the device includes multichannel miniflow Chip is controlled, the structure of the micro-fluidic chip includes being bonded to each other the substrate and cover plate of installing together, and the substrate and cover plate are equal For plate object or tablet, the groove via mould pressing process or etching technics formation is contained in that face towards the cover plate of the substrate Road structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or letter The window structure that single drilling technology is formed, it is bonded to each other the substrate being installed together and has been built into jointly containing pipe with the cover plate The micro-fluidic chip of road structure and the liquid pool structure being attached thereto, the locations of structures of the pipeline are located at the substrate and the cover plate phase The interface zone being mutually bonded, 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 institute The locations of structures of liquid pool is stated, the liquid pool there are 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 an one or more sample introduction end liquid pool, it is described enter Sample end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the terminal location of the micro-fluidic chip Then 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, one end of the pipeline and the sample introduction end liquid pool positioned at sample introduction end UNICOM, the other end of the pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip, and, sequentially Or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and reference electrode, the order Its locations of structures of the reference electrode is referred to closer to the terminal location, the backward refers to the reference electrode structure The sample introduction end position is located closer to, the working electrode is by conductive electrode and the bag being attached on the conductive electrode The gold size sensitive membrane for having buried tumor markers antibody is formed, and parallel construction is presented in the construction of the pipeline, and described is in parallel construction Pipeline is made up of six lateral parallel connections, and its appearance profile of the pipeline of the presentation parallel construction is similar to parallel circuit Profile, the quantity of the working electrode is six, and the installation position of six working electrodes is located at six branches respectively In pipeline, and, the tumor markers antibody in its top layer gold size sensitivity membrane structure of six working electrodes is to tumour respectively Six kinds of tumor markers antibody materials that mark antigen can be specifically bound, six kinds of antibody materials are tumor markers respectively Antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen are the antigen of broad sense, and the antibody is the anti-of broad sense Body, its material of the working electrode are argent material, gold material, carbon material or thermally decompose conducting polymer material, institute State working electrode its pattern and be presented column, sheet or thread, its material of the substrate is dimethyl silicone polymer material, the substrate 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, the structure of the device also include chain ring type magnetic holding device, the chain Share six links in its structure of ring type magnetic holding device, the link in strip, shaft-like or bar-shaped, mutually cut with scissors by each adjacent link Connect to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, its structure of each drift angle of the hexagonal annular structure Position is articulated structure position, and in the link of opposite side position, its material is magnetic conduction to two therein of the hexagonal annular structure Property material, its material of the remaining four processes of the hexagonal annular structure is non-magnetic material, what the non magnetic material referred to It is the material not responded strongly to externally-applied magnetic field, is wound with and encourages in a link among two links of the magnetic conductivity material Magnetic coil, the link for being wound with magnet exciting coil are respectively equiped with a magnetic conductivity material in the both sides neighbor positions of its magnet exciting coil The pole shoe of matter, two pole shoes are shared, two pole shoes are directed to the link of another magnetic conductivity material, another magnetic conduction Property its surface of the link of material on attach fixed chain ring type for being equiped with miniature ultrasonic transducer units, relying on electromagnetism adhesive strength Magnetic holding device is socketed the close position for being positioned at its terminal of the micro-fluidic chip, and, higher-order of oscillation electric signal transmission electricity Cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable link together；The chain ring type electromagnetism Fixture provides a function of facilitating the device to disassemble；Its major function of the miniature ultrasonic transducer units is in micro-fluidic chip During actual sample introduction test, the ultrasonic wave launched using it is opened to reduce the interface between sample solution and the inwall of the pipeline Power, can be compatible, also, using the sample introduction end and the terminal and the miniature ultrasonic transducer units installation position it Between distance difference and the difference on its ultrasonic intensity experienced, its interfacial tension of sample introduction end described in induced synthesis with Difference between its interfacial tension of the terminal, the interfacial tension difference between the micro-fluidic chip both ends can be micro-fluidic at this Pressure gap is formed between the both ends of chip, the pressure gap can drive sample solution to the end flow；This is miniature super The ultrasonic wave that acoustic wave transducer its function also includes being launched with it check large biological molecule contained in sample its described Absorption on inner surface of pipeline, and then check the substrate of the dimethyl silicone polymer material its body phase and the large biological molecule is gulped down Do not act on；Its function of the substrate of the dimethyl silicone polymer material is included with cover plate and working electrode and to electrode and reference electricity Pole together builds the micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function also include with Its property to the strong absorption of ultrasonic wave, ultrasonic wave is absorbed strongly, and thereby the micro-fluidic chip terminal to should The rapid decrement of ultrasonic intensity is realized within limited short distance between sample introduction end, it is high filled with some in the terminal liquid pool Water absorbing resin particle, its opening end of the terminal liquid pool are covered by breathable microporous film.

Only for the word of high hydroscopic resin one art-recognized meanings of itself, come for the professional of chemical field Say, be known.

The high hydroscopic resin is commercially available.

Only for the word of breathable microporous film one art-recognized meanings of itself, for the professional of technical field of membrane, It is known.

The breathable microporous film is commercially available.

The tumor markers antibody is the antibody of broad sense, and the antibody of the broad sense refers to possessing antibody function or functionally Can occur to combine with tumor markers involved by various corresponding clinics and form immune complex or exempt from similar to antibody The material of the analog of epidemic disease compound；The tumor markers antigen is the antigen of broad sense, and the antigen of the broad sense refers to can Using corresponding antibodies or the material for being functionally similar to antibody need to differentiate, detect involved by the various clinics of enzyme mark detection Tumor markers.

The word of magnet exciting coil one art-recognized meanings of itself are known.

The word of magnetic conductivity material one art-recognized meanings of itself are known；The material for belonging to magnetic conductivity material is for example pure Iron, silicon steel, soft magnetic ferrite etc..

Non magnetic material described in this case refers to the material not responded strongly to externally-applied magnetic field, belongs to the non magnetic material The material of matter such as metallic copper, metallic zinc, ormolu, metallic aluminium, almag etc..

The be hinged word art-recognized meanings of itself field in Machine Design is known.

The gold size sensitive membrane is to be sufficiently mixed chitosan gold size solution and tumor markers antibody-solutions uniformly, is used a little Sample instrument point sample is coated on specified structure position, and forms its drying and forming-film.Tumor markers in the gold size sensitive membrane Antibody is the tumor markers antibody of horseradish peroxidase or glucose oxidase mark, and the gold size sensitive membrane has been wrapped Containing introducing complementary medium therein for the above-mentioned each tumor markers antibody of fixation, the complementary medium such as chitosan, Cellulose acetate, gelatin be 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 any Selected size, still, for using prepare liquid sample less as far as possible and reducing the consideration of reagent loss etc., the pipeline includes The passage of the preferred capillary level of lateral, the passage of the capillary level imply that its internal diameter and the capillary on ordinary meaning The suitable passage of the internal diameter of pipe.The shape of cross section of its inner passage of capillary can be arbitrary shape, described transversal Face shape is for example circular, oval, square, rectangle, bar shaped, naturally it is also possible to be the linear of bending, also, institute arbitrarily be present The interior shape of capillary is stated with the extension of pipeline, the shape of cross section of different parts can also allow to be different shapes. Only for the word of capillary one, its art-recognized meanings is known.

What is be related in structure is the electrode of microsize to electrode and reference electrode, and its electrode shape, which may each be, appoints The selected shape of meaning, the arbitrarily selected shape such as column, sheet, strip or thread etc..It is described to electrode and institute The art-recognized meanings for stating the reference electrode vocabulary of itself are known.

It is related to several liquid pools in this case microfluidic chip structure, the liquid pool is the pond shape or capsule for transitional liquid storage Shape constructs, and its shape of the inner chamber of each liquid pool may each be arbitrarily selected shape, and the cavity shape is for example round Cylindrical cavity shape, square column type cavity-like, oblong cavity shape or spherical hollow space shape etc..

Only for professional of the word of ultrasonic transducer one art-recognized meanings of itself for ultrasonic technology field, It is known.

Various sizes, variously-shaped ultrasonic transducer are commercially available；Its size of commercially available miniature ultrasonic transducer units It may diminish to the magnitude only calculated with millimeter.

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

Only with regard to naked PDMS substrates itself micro-channel molding or lithographic technique for, be open-and-shut known skill Art；Similarly, the technology of hole-opening is even more known simple technique on naked PDMS substrates.This case is not also superfluous to this expansion Speech.

The industrial products market of involved its all size of higher-order of oscillation electric signal transmission cable is on sale.

The structure of the device can also include higher-order of oscillation electric signal generator；The higher-order of oscillation electric signal transmission cable Its other end can be connected with the higher-order of oscillation electric signal generator.

The involved higher-order of oscillation electric signal generator technology of itself, come for the professional in ultrasonic technology field Say, be simple and known；The higher-order of oscillation electric signal generator can customize to ultrasonic instrument specialized factory.

The preferred scope of its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units is between 5 milliwatts and 9000 milliwatts Between；The preferred scope of the frequency of its ultrasonic wave operationally launched of the miniature ultrasonic transducer units be between 100KHz with Between 12MHz.

This case device can further include some annexes certainly, and the annex is such as multiple tracks electrochemical workstation Deng the art-recognized meanings of the multiple tracks electrochemical workstation are known.The each work being related in this case microfluidic chip structure Electrode and to electrode and reference electrode etc., special it can get lines crossed and the multiple tracks electrochemical workstation via corresponding respectively The corresponding interface coupled.It is described that special to get lines crossed be for by each phase of each electrode and the multiple tracks electrochemical workstation Answer the private cable that interface is coupled to each other.The micro-fluidic chip in this case device, its structure can also include micro-valve, The quantity of the micro-valve is unlimited, and according to being actually needed, the micro-valve can be installed in any need in the microfluidic chip structure Position to be mounted；The word of micro-valve one is for the professional of micro fluidic chip technical field, the art-recognized meanings of itself It is known；The micro-valve itself manufacturing technology and the use of technology is also known；The component that the micro-valve is not required.

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

It is installed in by spraying or point sample instrument point sample or the coating of other appropriate process described in the working electrode surface layer Gold size sensitive membrane, its thicknesses of layers can allow be any setting treat sample measuring liquid occur electrical signals response thickness, It is however recommended to thickness preferable thickness is between 10 nanometers and 200 nanometers in other words.

The cover plate in chip structure, its material can allow to be any electrical insulating property material, such as：Polypropylene, Glass, polymethyl methacrylate, dimethyl silicone polymer, etc., in order to make smaller size of micro-fluidic chip, for example do Into the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and realized in the extremely short distance to ultrasonic wave Extremely fast decay, can preferably dimethyl silicone polymer be used as cover plate.Certainly, selected on large-sized micro-fluidic chip Using dimethyl silicone polymer it is used as the cover plate, and this case technical scheme is allowed.

The preferred scope at the distance between the terminal and the sample introduction end is between 3 centimetres and 10 centimetres.

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

The application method of this case micro-fluidic chip：

Itd is proposed first based on this case and the first public new liquid stream driving principle, its application of this case micro-fluidic chip are transported Among work, the new liquid stream driving method is determined completely without involving any additional Micropump.

The interfacial tension difference that this case is formed between micro-fluidic chip both ends caused by the ultrasonic wave, Driving liquid stream flows in the capillary channel of the six-channel microfluidic chip, right respectively using multi-channel electrochemical analyzer device Six kinds of classifiable tumor mark antigens carry out joint-detection.

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

1st, blood serum sample liquid is added in micro-pipe road, under ultrasonic wave driving, various tumor markers antigen molecules The tumor markers antibody of the corresponding horseradish peroxidase-labeled embedded by gold size sensitive membrane on electrode surface in each passage Capture.

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

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

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

It is an advantage of the invention that its close position relies on electromagnetism adhesive strength set in the terminal of the micro-fluidic chip The described chain ring type magnetic holding device of positioning is connect, is installed with institute's attaching in the chain ring type magnetic holding device one magnetic conductivity link micro- Type ultrasonic transducer, the low-power launched using it, the ultrasonic wave of high-frequency band so that without surface chemical modification or It is compatible between strong hydrophobic its tube wall of micro-fluidic chip internal pipeline and the test object aqueous solution of surface chemical modification Property be significantly increased, this is sample liquid stream by providing a realistic possibility；Meanwhile utilize dimethyl silicone polymer substrate Its strong absorbability to ultrasonic wave, in shorter distance, it is, from the terminal to the sample introduction end In the very short distance of only several centimeters yardsticks, reach the rapid decrement of ultrasonic intensity, thereby described in the micro-fluidic chip The difference of the interfacial tension is caused at both ends, and then, using the interfacial tension between the both ends difference its formed this two Pressure gap between end, driving sample liquid stream is in the strong hydrophobic capillary channel of such a script to the terminal side To flowing.By this case liquid stream drive scheme, entirely without must be to the substrate and its internal pipeline of the dimethyl silicone polymer material Any surface chemical modification or chemical modification are carried out, has altogether dispensed with the trouble journey of the surface chemical modification or chemical modification Sequence；And the equipment of traditional Micropump etc is altogether dispensed with；On the other hand, the low-power, the ultrasound of high-frequency band Ripple, additionally it is possible to contain large biological molecule in sample in literalness naked its inner surface of pipeline of dimethyl silicone polymer substrate On absorption, and then contain the swallow up effect of its body phase of dimethyl silicone polymer substrate to the large biological molecule；It is described anti- Former, antibody and the Reversible binding thing of antigen and antibody are all the type for belonging to described large biological molecule certainly；Due to described Suction-operated and the described effect of swallowing up effectively contained that therefore, dependence test result will be better able to objectively anti- Reflect actual conditions；The effect of the low-power, high-frequency band ultrasonic wave, also include facilitating the reversible knot between antigen, antibody certainly Close the quick of reaction to reach, this causes dependence test operation can be with than the completion of faster speed.

Due to surface chemical modification or chemistry for its relevant surfaces of the dimethyl silicone polymer substrate need not be carried out Modified operation, therefore, this surface chemical modification layer or chemically modified layer not need to exist, then, the poly dimethyl Its body phase interior polymer molecule of siloxanes substrate constantly diffuses to the surface automatically, migrate caused by it to the surface chemistry The damaging influence of decorative layer or chemically modified layer is also just not present.

It is related that the technical scheme of this case has dissolved its application to dimethyl silicone polymer substrate addressed above totally A series of technical barriers.Based on this case scheme, the very cheap polydimethyl siloxane material of this kind is just possible to micro- at this It is prepared by fluidic chip, production, using etc. field play bigger effect.

Attached in the chain ring type magnetic holding device one magnetic conductivity link in this case structure it is fixed installed it is described miniature Ultrasonic transducer, this architecture provides a function of facilitating the device to disassemble, in this way, the chain ring type magnetic holding device is together with it The miniature ultrasonic transducer units attached in one magnetic conductivity link just easily can be mutually disengaged with the micro-fluidic chip, that , the component that the part can freely depart from reuses many times in which just can circulate benignly；The architectural feature is advantageous to save The use cost of the device.

The framework of its large-scale integrated of this case micro-fluidic chip, it is determined among actual test application, to serum The requirement of sample is smaller, and this contributes to the body and mind damage for reducing related subject.

Brief description of the drawings

Fig. 1 be this case device chain ring type magnetic holding device its with appearance side rough under the micro-fluidic chip socket state View, the legend and indicates the relative position relation under the socket state between each component.

Fig. 2 is the outside side view under another viewing angle of this case device；Fig. 2 viewing angle and Fig. 1's Viewing angle is mutually perpendicular to.

In figure, 1 is chain ring type magnetic holding device, and 2 be the terminal of the micro-fluidic chip, and 3 be the institute of the micro-fluidic chip Sample introduction end is stated, 4 be the substrate of dimethyl silicone polymer material, and 5 be cover plate, and 6 be miniature ultrasonic transducer units, and 7 be the higher-order of oscillation Electric signal transmission cable, 8,15 respectively be positioned at different structure position two magnetic conductivity materials link, two magnetic conductivity The link of material end-to-end relation each other in the structure shown here, 9,22 be two pole shoes positioned at different structure position respectively, 10,12, 14th, 17,19,21 be six pin joints in six diverse locations respectively, and 11,13,18,20 be to be in four different positions respectively The link for the four non magnetic materials put, 16 be magnet exciting coil；The chain ring type magnetic holding device structure in legend is only signal Legend structure, the actual chain ring type magnetic holding device structure are not limited to the legend chain ring type magnetic holding device structure；Arrow in legend The symbology micro-fluidic chip its in actual motion, driven by pressure at two ends difference, the flow direction of its sample liquid stream.

Embodiment

In this case that Fig. 1 and Fig. 2 are shown embodiment, its main points of the example are that the structure of the device includes multichannel Micro-fluidic chip, the structure of the micro-fluidic chip include being bonded to each other the substrate 4 and cover plate 5 of installing together, the He of substrate 4 Cover plate 5 is plate object or tablet, and that face towards the cover plate 5 of the substrate 4 is contained via mould pressing process or etching work Skill formed channel structure, the substrate 4 also containing be connected with the channel structure and pierce the substrate 4 via mould pressing process, The window structure that etching technics or simple drilling technology are formed, is bonded to each other the substrate 4 being installed together and the cover plate 5 is common The micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto is built into, the locations of structures of the pipeline is located at should The interface zone that substrate 4 is bonded to each other with the cover plate 5, its side of the window is blocked by the cover plate 5 and opposite side opens, the window Locations of structures be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure respectively The sample introduction end liquid pool and terminal liquid pool of position, the position of sample introduction end 3 of the micro-fluidic chip have one or more described Sample introduction end liquid pool, the sample introduction end 3 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, micro- at this Then there is a terminal liquid pool position of terminal 2 of fluidic chip, and the terminal 2 refers to that the actual sample introduction of the micro-fluidic chip is surveyed The terminal location that liquid flows in its chip during examination, the terminal 2 are located remotely from each other with the sample introduction end 3, and one end of the pipeline is with being located at The sample introduction end liquid pool UNICOM at sample introduction end 3, the other end of the pipeline are described with the terminal 2 positioned at the micro-fluidic chip Terminal liquid pool UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and right Electrode and reference electrode, the order refers to its locations of structures of the reference electrode closer to the position of terminal 2, described Backward refers to the reference electrode locations of structures closer to the position of sample introduction end 3, and the working electrode is by conductive electrode And the gold size sensitive membrane for having embedded tumor markers antibody being attached on the conductive electrode is formed, the construction of the pipeline is in Existing parallel construction, the pipeline in parallel construction are made up of six lateral parallel connections, described that the described of parallel construction is presented Its appearance profile of pipeline is similar to the profile of parallel circuit, and the quantity of the working electrode is six, six working electrodes Installation position is located in six laterals respectively, and, in its top layer gold size sensitivity membrane structure of six working electrodes Tumor markers antibody be the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, should Six kinds of antibody materials are tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3 respectively, and the antigen is wide The antigen of justice, the antibody is the antibody of broad sense, and its material of the working electrode is argent material, gold material, carbon material Column, sheet or thread is presented in matter or thermal decomposition conducting polymer material, the working electrode its pattern, its material of the substrate 4 Dimethyl silicone polymer material, its surface of substrate 4 is the surface of primary form, the surface of the primary form its be meant to It is the surface of the primary form for the material for not passing through any surface chemical modification or any surface chemical modification, the device Structure also includes chain ring type magnetic holding device 1, and six links are shared in its structure of the chain ring type magnetic holding device 1, six links point It is not link 8,11,13,15,18,20, the link 8,11,13,15,18,20 is in strip, shaft-like or bar-shaped, each adjacent ring Section is hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, each top of the hexagonal annular structure Its locations of structures 10,12,14,17,19,21 of angle is articulated structure position, and two therein of the hexagonal annular structure is in Its material of the link 8,15 of opposite side position is magnetic conductivity material, the remaining four processes 11 of the hexagonal annular structure, 13,18, 20 its material are non-magnetic materials, and the non magnetic material refers to the material not responded strongly to externally-applied magnetic field, and this is led Magnet exciting coil 16 is wound with a link 15 among two links of magnetic material, this is wound with the link of magnet exciting coil 16 15 are respectively equiped with the pole shoe 9,22 of a magnetic conductivity material in the both sides neighbor positions of its magnet exciting coil 16, share described in two Pole shoe 9,22, two pole shoes 9,22 are directed to the link 8 of another magnetic conductivity material, the link of another magnetic conductivity material Fixed chain ring type magnetic holding device 1 for being equiped with miniature ultrasonic transducer units 6, relying on electromagnetism adhesive strength is attached on 8 its surface Socket is positioned at the close position of its terminal 2 of the micro-fluidic chip, and, higher-order of oscillation electric signal transmission cable 7, the height Frequency vibration swings one end of electric signal transmission cable 7 and the miniature ultrasonic transducer units 6 link together；The chain ring type magnetic holding device 1 Provide a function of facilitating the device to disassemble；Its major function of the miniature ultrasonic transducer units 6 is real in micro-fluidic chip During the sample introduction test of border, the ultrasonic wave launched using it is opened to reduce the interface between sample solution and the inwall of the pipeline Power, can be compatible, also, utilizes the sample introduction end 3 and the terminal 2 and the miniature ultrasonic transducer units 6 installing position The distance between put the difference on difference and its ultrasonic intensity experienced, its interface of sample introduction end 3 described in induced synthesis Difference between tension force and the terminal 2 its interfacial tension, the interfacial tension difference meeting between the micro-fluidic chip both ends 2,3 Pressure gap is formed between the both ends 2,3 of the micro-fluidic chip, the pressure gap can drive sample solution to the terminal 2 Flowing；Its function of miniature ultrasonic transducer units 6 also ultrasonic wave including being launched with it checks biology contained in sample Its absorption on the inner surface of pipeline of macromolecular, and then check the substrate 4 of the dimethyl silicone polymer material its body phase pair The effect of swallowing up of the large biological molecule；Its function of the substrate 4 of the dimethyl silicone polymer material includes and cover plate 5 and work electricity Pole and the micro-fluidic chip is together built to electrode and reference electrode, the soft dimethyl silicone polymer material for simultaneously having elasticity Its function of substrate 4 is also included with its property to the strong absorption of ultrasonic wave, and ultrasonic wave is absorbed strongly, and thereby micro- at this The fluidic chip terminal 2 is to the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end 3, the end Some high hydroscopic resin particles are filled with the liquid pool of end, its opening end of the terminal liquid pool is covered by breathable microporous film.

Arrow in legend indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, its try The flow direction of sample liquid stream.

It is attached that the higher-order of oscillation electric signal generator and multiple tracks electrochemical workstation etc. are depicted without in Fig. 1 and Fig. 2 Part.

Involved miniature ultrasonic transducer units 6 are commercially available；It can also be customized to ultrasonic transducer producer.

Involved higher-order of oscillation electric signal transmission cable 7 is commercially available；Can also be special to ultrasonic transducer producer or cable Industry producer customizes.

Involved higher-order of oscillation electric signal generator market has the product close to needs commercially available；It can also determine to relevant manufacturers System.

Each working electrode in this example structure and can be respectively via each special electricity to electrode and reference electrode Cable or say is being got lines crossed respectively with the corresponding cable interface of the multiple tracks electrochemical workstation as annex or saying interface connection of getting lines crossed.

The multiple tracks electrochemical workstation is commercially available；The multiple tracks electrochemical workstation can also be according to specific needs to phase Close specialized factory's customization.

In view of the pipeline of the presentation parallel construction its form foot that this case related text above expresses that it is described It is enough 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；Exempt from described in this case Epidemic disease compound refers to the immune complex of broad sense.

Claims

1. the tumor markers joint-detection six-channel microfluidic chip device simplified, 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, institute It is plate object or tablet to state substrate and cover plate, and that face towards the cover plate of the substrate was contained via mould pressing process or quarter Etching technique formed channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via stamper The window structure that skill, etching technics or simple drilling technology are formed, is bonded to each other the substrate being installed together and is total to the cover plate With the micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto has been built into, the locations of structures of the pipeline is 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, the window Locations of structures be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure respectively The sample introduction end liquid pool and terminal liquid pool of position, the sample introduction end position of the micro-fluidic chip have it is one or more it is described enter Sample end liquid pool, the sample introduction end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the miniflow The terminal location of control chip then has a terminal liquid pool, when the terminal refers to that the actual sample introduction of the micro-fluidic chip is tested The terminal location that liquid flows in its chip, the terminal are located remotely from each other with the sample introduction end, and one end of the pipeline is with being located at sample introduction end The sample introduction end liquid pool UNICOM, the other end of the pipeline with positioned at the micro-fluidic chip the terminal the terminal liquid pool UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and Reference electrode, the order refer to its locations of structures of the reference electrode closer to the terminal location, what the backward referred to Be the reference electrode locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached to The gold size sensitive membrane for having embedded tumor markers antibody on the conductive electrode is formed, and structure in parallel is presented in the construction of the pipeline Make, the pipeline in parallel construction is made up of six lateral parallel connections, and the pipeline that parallel construction is presented is outside it Shape profile is similar to the profile of parallel circuit, and the quantity of the working electrode is six, the installation position of six working electrodes Respectively in six laterals, and, the tumour mark in its top layer gold size sensitivity membrane structure of six working electrodes Will thing antibody is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, six kinds of antibody Material is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3 respectively, and the antigen is the anti-of broad sense Original, the antibody are the antibody of broad sense, and its material of the working electrode is argent material, gold material, carbon material or heat Conducting polymer material is decomposed, column, sheet or thread is presented in its pattern of the working electrode, and its material of the substrate is poly- diformazan Radical siloxane material, its surface of the substrate are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not pass through The surface of the primary form of the material of any surface chemical modification or any surface chemical modification is crossed, the structure of the device is also wrapped Include chain ring type magnetic holding device, share six links in its structure of the chain ring type magnetic holding device, the link in strip, it is shaft-like or Bar-shaped, each adjacent link is hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, the hexagonal annular Its locations of structures of each drift angle of structure is articulated structure position, and two therein of the hexagonal annular structure is in opposite side position Its material of the link put is magnetic conductivity material, and its material of the remaining four processes of the hexagonal annular structure is non magnetic material Matter, the non magnetic material refer to the material not responded strongly to externally-applied magnetic field, two links of the magnetic conductivity material it In a link on be wound with magnet exciting coil, this is wound with both sides neighbor positions of the link in its magnet exciting coil of magnet exciting coil On be respectively equiped with the pole shoe of a magnetic conductivity material, share two pole shoes, two pole shoes are directed to another magnetic conductivity The link of material, fixation is attached on its surface of the link of another magnetic conductivity material and is equiped with miniature ultrasonic transducer units, according to The chain ring type magnetic holding device socket of support electromagnetism adhesive strength is positioned at the close position of its terminal of the micro-fluidic chip, with And higher-order of oscillation electric signal transmission cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable Link together；The chain ring type magnetic holding device provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units Its major function is in the test of micro-fluidic chip actual sample introduction, and the ultrasonic wave launched using it reduces sample solution and institute The interfacial tension between the inwall of pipeline is stated, can be compatible, also, it is micro- with this using the sample introduction end and the terminal Difference in the distance between type ultrasonic transducer installation position difference and its ultrasonic intensity experienced, induce shape Difference between described its interfacial tension of sample introduction end and the terminal its interfacial tension, between the micro-fluidic chip both ends Interfacial tension difference can form pressure gap between the both ends of the micro-fluidic chip, and the pressure gap can drive sample solution To the end flow；The ultrasonic wave that its function of the miniature ultrasonic transducer units also includes being launched with it is checked contained in sample Its absorption on the inner surface of pipeline of some large biological molecules, so check the dimethyl silicone polymer material substrate its Swallow up effect of the body phase to the large biological molecule；Its function of the substrate of the dimethyl silicone polymer material includes and cover plate and work Make electrode and the micro-fluidic chip is together built to electrode and reference electrode, the dimethyl silicone polymer material that is soft and having elasticity Its function of the substrate of matter also includes with its property to the strong absorption of ultrasonic wave, and ultrasonic wave is absorbed strongly, and thereby exists The micro-fluidic chip terminal, should to the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end Some high hydroscopic resin particles are filled with terminal liquid pool, its opening end of the terminal liquid pool is covered by breathable microporous film.

2. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is that it is capillary channel that the pipeline, which includes the lateral,.

3. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, 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 tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, 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 is between 1 micron to 15000 microns.

5. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is that the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is, the cover plate its material in structure is dimethyl silicone polymer material.

7. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is that the distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.

8. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is that described its thickness of cover plate and substrate in structure is between 1.0 millimeters and 5.0 millimeters.

9. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is 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 electricity Its other end of cable is connected with the higher-order of oscillation electric signal generator.

10. the tumor markers joint-detection six-channel microfluidic chip device according to claim 1 simplified, it is special Sign is that for its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units between 5 milliwatts and 9000 milliwatts, this is miniature super The frequency of its ultrasonic wave operationally launched of acoustic wave transducer is between 100KHz and 12MHz.