CN107694645A

CN107694645A - Generally investigate ten Five-channel micro flow control chip devices of tumor markers

Info

Publication number: CN107694645A
Application number: CN201610703752.3A
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 one kind to generally investigate ten Five-channel micro flow control chip devices of tumor markers, belongs to analysis testing field.PDMS is substrate of the dimethyl silicone polymer as being used for making the kind tumor markers joint-detection micro-fluidic chip of general type more than ten, is had a clear superiority, but there is also some problems for needing to overcome；This case is directed to the 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, therefore 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 originally hydrophobic capillary channel to the terminal direction.The device exempts to use Micropump.

Description

Generally investigate ten Five-channel micro flow control chip devices of tumor markers

Technical field

The present invention relates to one kind to generally investigate ten Five-channel micro flow control chip devices of tumor markers, belongs to analysis test neck Domain.

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, by the solution be applied to build a kind of new crowd's physical examination is blanket being capable of pin Ten kinds are carried out with the micro flow control chip device of examination simultaneously while detection than more typical primary tumor mark.

The present invention solves the technical problem by following scheme, and the device that the program provides is a kind of generaI investigation tumor-marker Thing is that the structure of the device includes multichannel micro-fluidic chip, and this is micro- with ten Five-channel micro flow control chip devices, program main points The structure of fluidic chip includes being bonded to each other the substrate and cover plate of installing together, and the substrate and cover plate are plate object or piece The channel structure via mould pressing process or etching technics formation, the base are contained in shape thing, that face towards the cover plate of the substrate Piece also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or simple drilling technology shape Into window structure, be bonded to each other the substrate that is installed together and the cover plate be built into jointly containing pipeline configuration and with Connected liquid pool structure micro-fluidic chip, the locations of structures of the pipeline be located at the interface that the substrate and the cover plate are bonded to each other Region, its side of the window is blocked by the cover plate and opposite side opens, and the locations of structures of the window is exactly the structure of the liquid pool Position, the liquid pool have two kinds, and two kinds of liquid pools are the sample introduction end liquid pool and terminal liquid pool positioned at different structure position respectively, The sample introduction end position of the micro-fluidic chip has one or more sample introduction end liquid pool, and the sample introduction end refers to that this is micro- The injection end position of detected solution, then has an end in the terminal location of the micro-fluidic chip during fluidic chip actual sample introduction Liquid pool is held, the terminal refers to the terminal location of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, should Terminal is 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 pipeline The other end with positioned at the micro-fluidic chip the terminal the terminal liquid pool UNICOM, and, sequentially or backward respectively The working electrode that is installed in the pipeline on diverse location and the ginseng is referred to electrode and reference electrode, the order Than its locations of structures of electrode closer to the terminal location, the backward refers to the reference electrode locations of structures closer to institute State sample introduction end position, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumor-marker The gold size sensitive membrane of thing antibody is formed, and parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is by 15 Lateral is in parallel to be formed, and its appearance profile of the pipeline of the presentation parallel construction is similar to the profile of parallel circuit, institute The quantity for stating working electrode is 15, and the installation position of 15 working electrodes is located at 15 laterals respectively It is interior, and, the tumor markers antibody in its top layer gold size sensitivity membrane structure of 15 working electrodes is to tumour mark respectively 15 kinds of tumor markers antibody materials that will thing antigen can be specifically bound, 15 kinds of antibody materials are tumor-marker respectively Thing antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β- HCG, SCCA and β 2-MG, the antigen are the antigen of broad sense, and the antibody is the antibody of broad sense, its material of the working electrode Matter is argent material, gold material, carbon material or thermal decomposition conducting polymer material, and its pattern of the working electrode is presented Column, sheet or thread, its material of the substrate are dimethyl silicone polymer materials, and its surface of the substrate is the table of primary form Face, the surface of the primary form its be intended to refer to not by any surface chemical modification or any surface chemical modification should The surface of the primary form of material, the structure of the device also include chain ring type magnetic holding device, its structure of the chain ring type magnetic holding device In share six links, the link is in strip, shaft-like or bar-shaped, and each adjacent link is hinged to form chain link, the chain link The hexagonal cyclic structure of its appearance profile, its locations of structures of each drift angle of the hexagonal annular structure is articulated structure position Put, two therein of the hexagonal annular structure is magnetic conductivity material in the link of opposite side position its material, the hexagonal rings Its material of the remaining four processes of shape structure is non-magnetic material, and the non magnetic material refers to not strong to externally-applied magnetic field The material of strong response, magnet exciting coil is wound with a link among two links of the magnetic conductivity material, this, which is wound with, encourages The link of magnetic coil is respectively equiped with the pole shoe of a magnetic conductivity material in the both sides neighbor positions of its magnet exciting coil, shares two The pole shoe, two pole shoes are directed to the link of another magnetic conductivity material, its table of the link of another magnetic conductivity material The fixed chain ring type magnetic holding device socket positioning for being equiped with miniature ultrasonic transducer units, relying on electromagnetism adhesive strength is attached on face In the close position of its terminal of the micro-fluidic chip, and, higher-order of oscillation electric signal transmission cable, the higher-order of oscillation telecommunication One end of number transmission cable links together with the miniature ultrasonic transducer units；The chain ring type magnetic holding device provides a convenience The function that the device is disassembled；Its major function of the miniature ultrasonic transducer units is the profit in micro-fluidic chip actual sample introduction test The ultrasonic wave launched with it reduces the interfacial tension between sample solution and the inwall of the pipeline, can be compatible, Also, using the sample introduction end and the distance between the terminal and the miniature ultrasonic transducer units installation position difference and Difference on its ultrasonic intensity experienced, its interfacial tension of sample introduction end described in induced synthesis and its interface of the terminal Difference between power, the interfacial tension difference between the micro-fluidic chip both ends can be between the both ends of the micro-fluidic chip Pressure gap is formed, the pressure gap can drive sample solution to the end flow；Its function of the miniature ultrasonic transducer units Also include the ultrasonic wave launched with it and check contained its suction on the inner surface of pipeline of large biological molecule in sample It is attached, and then check swallow up effect of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule；Described poly- two Its function of the substrate of methylsiloxane material is micro- including building with cover plate and working electrode and together this to electrode and reference electrode Fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function also include it is strong to ultrasonic wave with it The property of absorption, ultrasonic wave is absorbed strongly, and thereby in micro-fluidic chip terminal to having between the sample introduction end The rapid decrement of ultrasonic intensity is realized within the short distance of limit, some high hydroscopic resin particles are filled with the terminal liquid pool, Its opening end of the terminal liquid pool 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.

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 ten Five-channels micro-fluidic chip, is distinguished using multi-channel electrochemical analyzer device Joint-detection is carried out to 15 kinds of tumor markerses.

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 of the corresponding horseradish peroxidase-labeled embedded by gold size sensitive membrane on electrode surface in each passage is special Property antibody capture.

2nd, the tumor markers specific antibody of horseradish peroxidase-labeled resists with the tumor markers in blood serum sample Original shape is into 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 15 lateral parallel connections, the institute that parallel construction is presented The profile that pipeline its appearance profile is similar to parallel circuit is stated, the quantity of the working electrode is 15, this 15 work The installation position of electrode respectively in 15 laterals, and, its top layer gold size of 15 working electrodes is quick The tumor markers antibody felt in membrane structure is the 15 kinds of tumor-markers that can be specifically bound to tumor markers antigen respectively Thing antibody materials, 15 kinds of antibody materials be respectively tumor markers antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, the antigen is 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 4 is poly- Dimethyl siloxane material, its surface of substrate 4 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 chain ring type magnetic holding device 1, share six links in its structure of the chain ring type magnetic holding device 1, six links are respectively 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 link phase It is mutually be hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, each drift angle of the hexagonal annular structure its Locations of structures 10,12,14,17,19,21 is articulated structure position, and two therein of the hexagonal annular structure is in opposite side Its material of the link 8,15 of position is magnetic conductivity material, the remaining four processes 11,13,18,20 of the hexagonal annular structure its Material is non-magnetic material, and the non magnetic material refers to the material not responded strongly to externally-applied magnetic field, the magnetic conductivity Magnet exciting coil 16 is wound with a link 15 among two links of material, the link 15 for being wound with magnet exciting coil 16 exists The pole shoe 9,22 of a magnetic conductivity material is respectively equiped with the both sides neighbor positions of its magnet exciting coil 16, shares two pole shoes 9th, 22, two pole shoes 9,22 are directed to the link 8 of another magnetic conductivity material, the link 8 of another magnetic conductivity material its Attach to fix on surface and be equiped with miniature ultrasonic transducer units 6, rely on the chain ring type magnetic holding device 1 of electromagnetism adhesive strength to be socketed The close position of its terminal 2 of the micro-fluidic chip is positioned at, and, higher-order of oscillation electric signal transmission cable 7, the high frequency vibrating The one end and the miniature ultrasonic transducer units 6 for swinging electric signal transmission cable 7 link together；The chain ring type magnetic holding device 1 provides One facilitates the function that the device is disassembled；Its major function of the miniature ultrasonic transducer units 6 be micro-fluidic chip it is actual enter When sample is tested, 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 3 and the terminal 2 and the installation position of miniature ultrasonic transducer units 6 Distance difference and its ultrasonic intensity experienced on difference, its interfacial tension of sample introduction end 3 described in induced synthesis with Difference between described its interfacial tension of terminal 2, the interfacial tension difference between the micro-fluidic chip both ends 2,3 can be micro- at this Pressure gap is formed between the both ends 2,3 of fluidic chip, the pressure gap can drive sample solution to be flowed to the terminal 2； Its function of miniature ultrasonic transducer units 6 also ultrasonic wave including being launched with it checks large biological molecule contained in sample Its absorption on the inner surface of pipeline, and then check the substrate 4 of the dimethyl silicone polymer material its body phase to the biology The effect of swallowing up of macromolecular；The substrate 4 of the dimethyl silicone polymer material its function includes and cover plate 5 and working electrode and right Electrode and reference electrode together build the micro-fluidic chip, the substrate 4 of the dimethyl silicone polymer material that is soft and having elasticity Its function is also included with its property to the strong absorption of ultrasonic wave, and ultrasonic wave is absorbed strongly, and thereby micro-fluidic at this The 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 terminal liquid Some high hydroscopic resin particles are filled with pond, 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. generally investigate ten Five-channel micro flow control chip devices of tumor markers, it is characterised in that the structure of the device includes more logical Road 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 substrate and Cover plate is plate object or tablet, and that face towards the cover plate of the substrate is contained via mould pressing process or etching technics shape Into channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etch work The window structure that skill or simple drilling technology are formed, it is bonded to each other the substrate being installed together and is built into jointly with the cover plate Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline are located at the substrate with being somebody's turn to do The interface zone that cover plate is bonded to each other, its side of the window is blocked by the cover plate and opposite side opens, the locations of structures of the window It is exactly the locations of structures of the liquid pool, the liquid pool has two kinds, and two kinds of liquid pools are the sample introduction positioned at different structure position respectively End liquid pool and terminal liquid pool, the sample introduction end position of the micro-fluidic chip have one or more sample introduction end liquid pool, The sample introduction end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, at the end of the micro-fluidic chip End position then has a terminal liquid pool, and the terminal refers to during the actual sample introduction test of the micro-fluidic chip liquid in its chip The terminal location of body flowing, the terminal are located remotely from each other with the sample introduction end, one end of the pipeline and the sample introduction positioned at 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 are held, and, Sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and reference electrode, institute State order and refer to its locations of structures of the reference electrode closer to the terminal location, the backward refers to the reference electricity For pole locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached to the conductive 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 in parallel connection The pipeline of construction is made up of 15 lateral parallel connections, and its appearance profile of the pipeline of the presentation parallel construction is similar to The profile of parallel circuit, the quantity of the working electrode is 15, and the installation position of 15 working electrodes is located at respectively In 15 laterals, and, the tumor markers in its top layer gold size sensitivity membrane structure of 15 working electrodes Antibody is the 15 kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, 15 kinds of antibody Material be respectively tumor markers antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, the antigen are the antigen of broad sense, and the antibody is broad sense Antibody, its material of the working electrode be argent material, gold material, carbon material or thermal decomposition conducting polymer material, The working electrode its pattern is presented column, sheet or thread, and 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 The surface of the primary form of the material of any surface chemical modification, the structure of the device also include chain ring type magnetic holding device, should Share six links in its structure of chain ring type magnetic holding device, for the link in strip, shaft-like or bar-shaped, each adjacent link is mutual It is be hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, its knot of each drift angle of the hexagonal annular structure Structure position is articulated structure position, and in the link of opposite side position, its material is to lead to two therein of the hexagonal annular structure Magnetic material, its material of the remaining four processes of the hexagonal annular structure is non-magnetic material, and the non magnetic material refers to Be the material not responded strongly to externally-applied magnetic field, be wound with a link among two links of the magnetic conductivity material Magnet exciting coil, the link for being wound with magnet exciting coil are respectively equiped with a magnetic conductivity in the both sides neighbor positions of its magnet exciting coil The pole shoe of material, share two pole shoes, two pole shoes are directed to the link of another magnetic conductivity material, this another lead Fixed chain link for being equiped with miniature ultrasonic transducer units, relying on electromagnetism adhesive strength is attached on its surface of the link of magnetic material Formula magnetic holding device is socketed the close position for being positioned at its terminal of the micro-fluidic chip, and, the transmission of higher-order of oscillation electric signal Cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable link together；Chain ring type electricity Magnetic holding device provides a function of facilitating the device to disassemble；Its major function of the miniature ultrasonic transducer units is in micro-fluidic core During the test of piece actual sample introduction, the ultrasonic wave launched using it reduces the interface between sample solution and the inwall of the pipeline Tension force, can be compatible, also, utilizes the sample introduction end and the terminal and the miniature ultrasonic transducer units installation position The distance between difference on difference and its ultrasonic intensity experienced, its interfacial tension of sample introduction end described in induced synthesis With the difference between the terminal its interfacial tension, the interfacial tension difference between the micro-fluidic chip both ends can be in the miniflow Pressure gap is formed between the both ends of control chip, the pressure gap can drive sample solution to the end flow；This is miniature The ultrasonic wave that ultrasonic transducer its function also includes being launched with it check large biological molecule contained in sample its in institute The absorption on inner surface of pipeline is stated, and then checks the substrate of the dimethyl silicone polymer material its body phase to the large biological molecule The effect of swallowing up；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 together builds the micro-fluidic chip, soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include With its property to the strong absorption of ultrasonic wave, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip, the terminal arrives The rapid decrement of ultrasonic intensity is realized within limited short distance between the sample introduction end, is filled with the terminal liquid pool High hydroscopic resin particle, its opening end of the terminal liquid pool are covered by breathable microporous film.

2. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that institute It is capillary channel to state pipeline to include the lateral.

3. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that institute It is the conductive material formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated to state thermal decomposition conducting polymer.

4. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that institute State working electrode width or diameter between 0.1 micron to 2000 microns, and, the length of the working electrode is micro- 1 Rice is between 15000 microns.

5. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that institute The thickness of gold size sensitive membrane is stated between 10 nanometers and 200 nanometers.

6. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that knot The cover plate its material in structure is dimethyl silicone polymer material.

7. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that institute The distance between terminal and the sample introduction end are stated between 3 centimetres and 10 centimetres.

8. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that knot Described its thickness of cover plate and substrate in structure is between 1.0 millimeters and 5.0 millimeters.

9. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that should The structure of micro fluidic device also includes higher-order of oscillation electric signal generator, its other end of the higher-order of oscillation electric signal transmission cable It is connected with the higher-order of oscillation electric signal generator.

10. generaI investigation ten Five-channel micro flow control chip devices of tumor markers according to claim 1, it is characterised in that Between 5 milliwatts and 9000 milliwatts, the miniature ultrasonic changes its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units The frequency of its ultrasonic wave operationally launched of energy device is between 100KHz and 12MHz.