CN107694646A

CN107694646A - Typical six kinds of tumor markers joint-detection micro flow control chip devices

Info

Publication number: CN107694646A
Application number: CN201610703755.7A
Authority: CN
Inventors: 洪小女
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 typical six kinds of tumor markers joint-detection micro flow control chip devices, 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, also have problem；This case is directed to the problem.This case main points are, the selected PDMS with ecosystem surface of substrate, and by the elastic clip that miniature ultrasonic transducer units are attached on its clamping limb with elastic force snap-in location in its neighbor positions of the sample liquid stream terminal of the micro-fluidic chip, interfacial tension is reduced with ultrasonic wave, the interface compatibility between 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 strong hydrophobic capillary channel originally to terminal direction.The device exempts to use Micropump.

Description

Typical six kinds of tumor markers joint-detection micro flow control chip devices

Technical field

The present invention relates to a kind of typical six kinds of tumor markers joint-detection micro flow control chip devices, belong to analysis test 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 typical six kinds of tumours Markers in detecting micro flow control chip device, the program are characterized in that 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 and cover plate of installing together, and the substrate and cover plate are plate The conduit knot via mould pressing process or etching technics formation is contained in shape thing or tablet, that face towards the cover plate of the substrate Structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or simply beat The window structure that hole technique is formed, it is bonded to each other the substrate being installed together and has been built into jointly with the cover plate and contain pipeline knot The micro-fluidic chip of structure and the liquid pool structure being attached thereto, the locations of structures of the pipeline are located at the substrate and mutually pasted with the cover plate The interface zone of conjunction, its side of the window is blocked by the cover plate and opposite side opens, and the locations of structures of the window is exactly the liquid The locations of structures in pond, the liquid pool have two kinds, two kinds of liquid pools respectively be positioned at different structure position sample introduction 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, the sample introduction end The injection end position of detected solution during the micro-fluidic chip actual sample introduction is referred to, is then had in the terminal location of the micro-fluidic chip One terminal liquid pool, the terminal refer to the end of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested End 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, The other end of the pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip, and, it is sequentially or inverse The working electrode that is respectively installed in sequence 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 Closer to the sample introduction end position, the working electrode is by conductive electrode and the embedding being attached on the conductive electrode The gold size sensitive membrane of tumor markers antibody is formed, and parallel construction, the pipeline in parallel construction is presented in the construction of the pipeline It is made up of six lateral parallel connections, its appearance profile of the pipeline of the presentation parallel construction is similar to the wheel of parallel circuit Exterior feature, the quantity of the working electrode is six, and the installation position of six working electrodes is located at six laterals respectively It is interior, and, the tumor markers antibody in its top layer gold size sensitivity membrane structure of six working electrodes is to tumor-marker respectively Six kinds of tumor markers antibody materials that thing antigen can be specifically bound, six kinds of antibody materials are tumor markers antibody respectively AFP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen are the antigen of broad sense, and the antibody is the antibody of broad sense, institute It is argent material, gold material, carbon material or thermal decomposition conducting polymer material to state working electrode its material, the work Column, sheet or thread is presented in its pattern of electrode, and its material of the substrate is dimethyl silicone polymer material, and its surface of the substrate is The surface of primary form, the surface of the primary form its be intended to refer to and do not pass through any surface chemical modification or any surface The surface of the primary form of the material of chemical modification, the structure of the device also include elastic clip, and two of the elastic clip are opposite Clamping limb snap-in location in the position of the neighbouring terminal of the micro-fluidic chip, a clamping limb at least in Upper attaching fixation is equiped with miniature ultrasonic transducer units, and, higher-order of oscillation electric signal transmission cable, the higher-order of oscillation electric signal One end of transmission cable links together with the miniature ultrasonic transducer units；The elastic clip provides one and facilitates the device to disassemble Function；Its major function of the miniature ultrasonic transducer units is in the actual sample introduction test of micro-fluidic chip, is launched using it Ultrasonic wave reduce the interfacial tension between sample solution and the inwall of the pipeline, can be compatible, also, utilize institute State the distance between sample introduction end and the terminal and miniature ultrasonic transducer units installation position difference and it is experienced Ultrasonic intensity on difference, the difference between its interfacial tension of sample introduction end described in induced synthesis and the terminal its interfacial tension Different, the interfacial tension difference between the micro-fluidic chip both ends can form pressure differential between the both ends of the micro-fluidic chip Different, the pressure gap can drive sample solution to the end flow；Its function of the miniature ultrasonic transducer units also includes with it The ultrasonic wave launched checks in sample contained its absorption on the inner surface of pipeline of large biological molecule, and then checks Swallow up effect of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule；The dimethyl silicone polymer material Its function of the substrate of matter includes with cover plate and working electrode and together builds the micro-fluidic chip to electrode and reference electrode, soft It is right and the substrate of the dimethyl silicone polymer material its function of having elasticity also includes with its property to the strong absorption of ultrasonic wave Ultrasonic wave is absorbed strongly, and thereby within micro-fluidic chip terminal to the limited short distance between the sample introduction end Realize the rapid decrement of ultrasonic intensity, be filled with some high hydroscopic resin particles in the terminal liquid pool, the terminal liquid pool its open Mouth end is 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.

Described miniature ultrasonic transducer units can certainly be all installed on two clamping limbs；But only install one Miniature ultrasonic transducer units are dealt with and used enough.

The word of elastic clip one art-recognized meanings of itself are 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 5000 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 in elastic force described in the terminal of the micro-fluidic chip its close position snap-in location Folder, with the clamping limb of the elastic clip institute attaching install miniature ultrasonic transducer units, the low-power launched using it, high frequency The ultrasonic wave of frequency range so that without inside strong hydrophobic micro-fluidic chip of surface chemical modification or surface chemical modification Compatibility between its tube wall of pipeline and the test object aqueous solution is significantly increased, and this is existing by providing one for sample liquid stream Real possibility；Meanwhile using its strong absorbability to ultrasonic wave of dimethyl silicone polymer substrate, in shorter distance It is interior, it is, in from the terminal to the very short distance of only several centimeters yardsticks the sample introduction end, it is strong to reach ultrasonic wave The rapid decrement of degree, the difference of the interfacial tension is thereby caused at the both ends of the micro-fluidic chip, and then, using this two Pressure gap between its both ends for being formed of the difference of interfacial tension between end, driving sample liquid stream is in such a original Flowed in this strong hydrophobic capillary channel to the terminal direction.By this case liquid stream drive scheme, entirely without must be to this The substrate and its internal pipeline of dimethyl silicone polymer material carry out any surface chemical modification or chemical modification, altogether dispense with The laborious procedures of the surface chemical modification or chemical modification；And setting for traditional Micropump etc is altogether dispensed with It is standby；On the other hand, the ultrasonic wave of the low-power, high-frequency band, additionally it is possible to contain the large biological molecule in sample at this without modification Naked its inner surface of pipeline of dimethyl silicone polymer substrate on absorption, and then contain its body of the dimethyl silicone polymer substrate The effect of swallowing up of relatively described large biological molecule；The Reversible binding thing of the antigen, antibody and antigen and antibody is all certainly Belong to the type of described large biological molecule；Because described suction-operated and the described effect of swallowing up effectively are contained, Therefore, dependence test result will be better able to objectively reflect actual conditions；The effect of the low-power, high-frequency band ultrasonic wave, Certainly also include facilitating the Reversible binding between antigen, antibody react it is quick reach, this causes dependence test operates can be with Completed than 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.

The miniature ultrasonic transducer units have been installed in fixation on the elastic clip its clamping limb in this case structure, and the structure carries A function of facilitating the device to disassemble is supplied, in this way, the elastic clip just can together with miniature ultrasonic transducer units appended thereon It is enough to be easily mutually disengaged with the micro-fluidic chip, then, the component that the part can freely depart from just can circulate benignly it is heavy It is multiple to use many times；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 is its rough outside side view of this case device.

In figure, 1 is elastic clip, and 2,7 be two clamping limbs of elastic clip respectively, and 3 be the terminal of the micro-fluidic chip, 4 be the sample introduction end of the micro-fluidic chip, and 5 be the substrate of dimethyl silicone polymer material, and 6 be cover plate, and 8 be miniature ultrasonic Transducer, 9 be higher-order of oscillation electric signal transmission cable, and 10 be tension spring；The spring clamp structure in legend is only signal Legend structure, actual spring clamp structure are not limited to the legend spring clamp structure；Arrow in legend indicates the micro-fluidic core Piece its in actual motion, by pressure at two ends difference drive, the flow direction of its sample liquid stream.

Embodiment

In this case that Fig. 1 is 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 5 and cover plate 6 of installing together, the substrate 5 and cover plate 6 It is plate object or tablet, that face towards the cover plate 6 of the substrate 5 is contained to be formed via mould pressing process or etching technics Channel structure, the substrate 5 also containing be connected with the channel structure and pierce the substrate 5 via mould pressing process, etch work The window structure that skill or simple drilling technology are formed, is bonded to each other the substrate 5 being installed together and is built into jointly with the cover plate 6 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 5 The interface zone being bonded to each other with the cover plate 6, its side of the window is blocked by the cover plate 6 and opposite side opens, the knot of the window Structure position is exactly the locations of structures of the liquid pool, and the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure position respectively Sample introduction end liquid pool and terminal liquid pool, there is the one or more sample introduction position of sample introduction end 4 of the micro-fluidic chip Liquid pool is held, the sample introduction end 4 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, micro-fluidic at this Then there is a terminal liquid pool position of terminal 3 of chip, when the terminal 3 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 3 are located remotely from each other with the sample introduction end 4, and one end of the pipeline is with being located at sample introduction The sample introduction end liquid pool UNICOM at end 4, the other end of the pipeline and the terminal of the terminal 3 positioned at the micro-fluidic chip 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 position of terminal 3, the backward The reference electrode locations of structures is referred to closer to the position of sample introduction end 4, the working electrode by conductive electrode and The gold size sensitive membrane for having embedded tumor markers antibody being attached on the conductive electrode is formed, and the construction of the pipeline is presented simultaneously Connection construction, the pipeline in parallel construction are made up of six lateral parallel connections, the pipeline that parallel construction is presented Its appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is six, the installing of six working electrodes Position is located in six laterals respectively, and, it is swollen in its top layer gold size sensitivity membrane structure of six working electrodes Tumor markers antibody is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, this six kinds Antibody materials are tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3 respectively, and the antigen is broad sense Antigen, the antibody are the antibody of broad sense, its material of the working electrode be argent material, gold material, carbon material or Conducting polymer material is thermally decomposed, column, sheet or thread is presented in the working electrode its pattern, and its material of the substrate 5 is poly- Dimethyl siloxane material, its surface of substrate 5 are the surfaces of primary form, the surface of the primary form its be intended to refer to not There are the surface of the primary form by any surface chemical modification or the material of any surface chemical modification, the structure of the device Also include elastic clip 1, two opposite clamping limbs 2 of the elastic clip 1,7 snap-in locations are in the neighbouring described of the micro-fluidic chip The position of terminal 3, a clamping limb at least in for example attaches on clamping limb 7 fixed to be equiped with miniature ultrasonic and changes Can device 8, and, higher-order of oscillation electric signal transmission cable 9, one end of the higher-order of oscillation electric signal transmission cable 9 is miniature super with this Acoustic wave transducer 8 links together；The elastic clip 1 provides a function of facilitating the device to disassemble；The miniature ultrasonic changes Its major function of energy device 8 is that the ultrasonic wave launched using it is molten to reduce sample in the actual sample introduction test of micro-fluidic chip Interfacial tension between the inwall of liquid and the pipeline, can be compatible, also, utilizes the sample introduction end 4 and the end Hold on the distance between 3 and the installation position of miniature ultrasonic transducer units 8 difference and its ultrasonic intensity experienced Difference between difference, its interfacial tension of sample introduction end 4 described in induced synthesis and the terminal 3 its interfacial tension, the micro-fluidic core Interfacial tension difference between the piece both ends 3,4 can form pressure gap between the both ends 3,4 of the micro-fluidic chip, the pressure Power difference can drive sample solution to be flowed to the terminal 3；Its function of the miniature ultrasonic transducer units 8 also includes being launched with it Ultrasonic wave check in sample contained its absorption on the inner surface of pipeline of large biological molecule, and then check this poly- two Swallow up effect of its body phase of the substrate 5 of methylsiloxane material to the large biological molecule；The dimethyl silicone polymer material Its function of substrate 5 includes with cover plate 6 and working electrode and electrode and reference electrode is together built the micro-fluidic chip, and softness is simultaneously Its function of the substrate 5 of the dimethyl silicone polymer material of tool elasticity also includes with its property to the strong absorption of ultrasonic wave, right Ultrasonic wave is absorbed strongly, and the limited short distance thereby arrived between the sample introduction end 4 in the micro-fluidic chip terminal 3 it Inside realize the rapid decrement of ultrasonic intensity, be filled with some high hydroscopic resin particles in the terminal liquid pool, the terminal liquid pool its Opening end 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.

The associate members such as the higher-order of oscillation electric signal generator and multiple tracks electrochemical workstation are depicted without in Fig. 1.

Itself is commercially available for involved elastic clip.It is available for the commercially available alternative spring clamp structure that this case structure uses And moulding and size etc., quality is various, and specific elastic clip pattern can be selected as needed.

Involved miniature ultrasonic transducer units 8 are commercially available；It can also be customized to ultrasonic transducer producer.

Involved higher-order of oscillation electric signal transmission cable 9 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. typical six kinds of tumor markers joint-detection micro flow control chip devices, it is characterised in 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 and cover plate of installing together, the base Piece 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, quarter The window structure that etching technique or simple drilling technology are formed, is bonded to each other the substrate being installed together and is built jointly with the cover plate Into the micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline is located at the substrate The interface zone being bonded to each other with the cover plate, its side of the window is blocked by the cover plate and opposite side opens, the structure of the window Position is exactly the locations of structures of the liquid pool, and the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure position respectively Sample introduction 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 Pond, the sample introduction end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the micro-fluidic chip Terminal location then have a terminal liquid pool, the terminal refers to its chip during the actual sample introduction test of the micro-fluidic chip The terminal location of interior liquid flowing, the terminal are located remotely from each other with the sample introduction end, and one end of the pipeline is with being located at the described of sample introduction end Sample introduction end liquid pool 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 electricity Pole, the order refer to its locations of structures of the reference electrode closer to the terminal location, and the backward refers to described For reference electrode locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached to the conduction Property electrode on the gold size sensitive membrane for having embedded tumor markers antibody form, parallel construction is presented in the construction of the pipeline, described It is made up of in the pipeline of parallel construction six lateral parallel connections, its appearance profile of the pipeline of the presentation parallel construction is near The profile of parallel circuit is similar to, the quantity of the working electrode is six, and the installation position of six working electrodes is located at respectively In six laterals, and, the tumor markers antibody in its top layer gold size sensitivity membrane structure of six working electrodes It is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, six kinds of antibody materials difference It is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen is the antigen of broad sense, described anti- Body is the antibody of broad sense, and its material of the working electrode is that argent material, gold material, carbon material or thermal decomposition are conductive high Column, sheet or thread is presented in molecule material, its pattern of the working electrode, and its material of the substrate is dimethyl silicone polymer material Matter, 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 any surface The surface of the primary form of the material of chemical modification or any surface chemical modification, the structure of the device also include elastic clip, Two opposite clamping limb snap-in locations of the elastic clip are in the position of the neighbouring terminal of the micro-fluidic chip, at least at it In a clamping limb on attach fixation be equiped with miniature ultrasonic transducer units, and, the 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 elastic clip provides One facilitates the function that the device is disassembled；Its major function of the miniature ultrasonic transducer units is in the actual sample introduction of micro-fluidic chip During test, the ultrasonic wave launched using it reduces the interfacial tension between sample solution and the inwall of the pipeline, makes it Can be compatible, also, using between the sample introduction end and the terminal and the miniature ultrasonic transducer units installation position away from Deviation is different and its ultrasonic intensity for being experienced on difference, its interfacial tension of sample introduction end described in induced synthesis and the end The difference between its interfacial tension is held, the interfacial tension difference between the micro-fluidic chip both ends can be in the micro-fluidic chip Pressure gap is formed between the both ends, the pressure gap can drive sample solution to the end flow；The miniature ultrasonic changes Can device its function ultrasonic wave for also including being launched with it check large biological molecule contained in sample its in the pipeline Absorption on surface, and then check swallow up work of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule With；Its function of the substrate of the dimethyl silicone polymer material is included with cover plate and working electrode and to electrode and reference electrode one With building the micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function also include it is right with its The property that ultrasonic wave absorbs strongly, ultrasonic wave is absorbed strongly, and thereby in micro-fluidic chip terminal to the sample introduction The rapid decrement of ultrasonic intensity is realized within limited short distance between end, some high water absorptions are filled with the terminal liquid pool Resin particle, its opening end of the terminal liquid pool are covered by breathable microporous film.

2. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In it is capillary channel that the pipeline, which includes the lateral,.

3. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In the thermal decomposition conducting polymer is the conductive material formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated.

4. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the length of the working electrode Between 1 micron to 15000 microns.

5. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In the cover plate its material in structure is dimethyl silicone polymer material.

7. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In the distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.

8. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In described its thickness of cover plate and substrate in structure is between 1.0 millimeters and 5.0 millimeters.

9. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist Also include higher-order of oscillation electric signal generator in the structure of, the micro fluidic device, the higher-order of oscillation electric signal transmission cable its The other end is connected with the higher-order of oscillation electric signal generator.

10. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, its feature exist In its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units is between 5 milliwatts and 5000 milliwatts, the miniature ultrasonic The frequency of its ultrasonic wave operationally launched of transducer is between 100KHz and 12MHz.