CN107694619A - Ten Five-channel micro flow control chip devices of common tumor markers examination - Google Patents

Abstract

The present invention relates to a kind of ten Five-channel micro flow control chip devices of common tumor markers examination, belong 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, advantageous, also there is 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, 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, the difference provides a kind of strength driven sample liquid stream and flowed along hydrophobic capillary channel to the terminal direction, the strength while with Micropump its mechanicalness pumping strength Collaboration for being included in structure.

Description

Ten Five-channel micro flow control chip devices of common tumor markers examination

Technical field

The present invention relates to a kind of ten Five-channel micro flow control chip devices of common tumor markers examination, belong to analysis and survey Examination 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.

Third, as described above, the PDMS micro-channel inner surfaces are strongly hydrophobic, and targetedly surface chemical modification or surface Chemical modification is difficult to persistently again, therefore, actually can only still belong to effective in the surface state after the modification of its surface or surface modification It is short-term within use it；If having had been subjected to that ofer short duration period of validity, and still by force if use, due to table Again close to hydrophobic state, bigger flowing is then already certainly existed using usual Micropump driving sample liquid stream for surface state Resistance, so, just by increasing Micropump pump power and pumping pressure test liquid can only be forced to flow to target direction flowing, as above Described, this PDMS material is soft, pumps sample liquid stream with too high, mechanical pumping pressure, will cause the substrate Bubbling, expanded, distortion, deformation, also, this high pressure feelings occur for the micro-channel that its sample introduction end includes sample introduction end near zone Under condition, the stripping between substrate and cover plate also easily occurs in the sample introduction end and its micro-channel of near zone and its periphery, Under this situation, the crack for the appearance that sample solution will enter after the stripping between the substrate and cover plate formed and it is horizontal everywhere Stream, this actually results in the damage of the micro-fluidic chip；It is certainly, not in place if surface modification or surface modification script, It can cause said circumstances occur within the of short duration usual term of validity；The feelings of liquid stream driving are being carried out using additional Micropump merely Under condition, the above-mentioned problem exists all the time.As described above, if any surface modification or surface modification etc. were not done completely Pre action, then, the above-mentioned problem will be even more serious, is roused even without micro-channel described in sample introduction end and its near zone occurs The problems such as being peeled off between bubble, expanded, distortion, deformation and substrate and cover plate, merely because the flow resistance is excessive, using high pressure Micropump also may not can drive sample liquid stream to be marched forward towards terminal side.

The content of the invention

The technical problem to be solved by the invention is to provide a package solution, while solve to address above Three aspect a series of problem actually mutually involved together, also, by the solution be applied to structure one Kind new crowd's physical examination is blanket can be directed to ten kinds and carries out examination simultaneously, same than more typical primary tumor mark When the micro flow control chip device that detects.

The present invention solves the technical problem by following scheme, and the device that the program provides is a kind of kinds of tumor mark Ten Five-channel micro flow control chip devices of thing examination, program main points are that the structure of the device includes micro-fluidic chip, and this is micro- 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 is contained in shape thing, that face towards the cover plate of the substrate, mutually The substrate being jointly mounted to together has been built into the micro-fluidic chip containing pipeline configuration, the knot of the pipeline jointly with the cover plate Structure position is located at the juncture area that the substrate and the cover plate are bonded to each other, and the both ends of the pipeline are entered with the micro-fluidic chip respectively Sample end and terminal connection, the sample introduction end is the injection end of the micro-fluidic chip sample solution, and the terminal is the micro-fluidic chip The terminal that sample solution flows in its chip during actual sample introduction test, the terminal are located remotely from each other with the sample introduction end, and the terminal is with being somebody's turn to do Between 3 centimetres and 10 centimetres, the terminal links together with terminal liquid pool at the distance between sample introduction end, the terminal liquid pool its The profile of structure is in pond shape, pit shape or cup-shaped, and the terminal liquid pool is used to receive the sample for flowing to terminal via the pipeline molten Liquid, it is, described in the pipeline on diverse location in order or backward is equiped with working electrode and to electrode and reference electrode Order refers to that its locations of structures of the reference electrode refers to the reference electrode closer to the terminal location, the backward For locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached on the conductive electrode The gold size sensitive membrane for having embedded tumor markers antibody form, parallel construction is presented in the construction of the pipeline, and described is in structure in parallel The pipeline made is made up of 15 lateral parallel connections, and its appearance profile of the pipeline of the presentation parallel construction is similar to simultaneously Join the profile of circuit, the quantity of the working electrode is 15, and the installation position of 15 working electrodes is located at institute respectively State in 15 laterals, and, the tumor markers in its top layer gold size sensitivity membrane structure of 15 working electrodes resists Body is the 15 kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, 15 kinds of antibody things Matter is tumor markers antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50, NSE, CYFRA21- respectively 1st, 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, 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 its inner passage of micro-fluidic chip, can phase Hold, also, utilize the sample introduction end and the distance between the terminal and the miniature ultrasonic transducer units installation position difference And the difference on its ultrasonic intensity for being experienced, its interfacial tension of sample introduction end described in induced synthesis and its boundary of the terminal Difference between the tension force of face, the interfacial tension difference between the micro-fluidic chip both ends can be at the both ends of the micro-fluidic chip Between form pressure gap, the pressure gap can drive sample solution to the end flow；The miniature ultrasonic transducer units its Function also include the ultrasonic wave launched with it check large biological molecule contained in sample its in the micro-fluidic chip in it Absorption on portion's channel inner surface, and then check the substrate of the dimethyl silicone polymer material its body phase to the large biological molecule The effect of swallowing up；It is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and include with it to the strong suction of ultrasonic wave The property of receipts, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip terminal to limited between the sample introduction end Short distance within realize the rapid decrement of ultrasonic intensity；And Micropump, the Micropump are connected with the sample introduction end；The Micropump Function is that the interfacial tension between the inwall of the sample solution and its inner passage of micro-fluidic chip is by the ul-trasonic irradiation And reduce, under the increased precondition of alternate compatibility, come and ultrasonic wave induction with the mechanicalness pumping strength of the Micropump Interfacial tension difference its caused driving force between the both ends is supported mutually, adjusts mutually, is mutually coupled, with collaboration The mode of running pools the strength that one driving test liquid flows to the terminal direction flowing, and one is filled with the terminal liquid pool A little high hydroscopic resin particles, its opening end of the terminal liquid pool are covered by breathable microporous film.

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

The high hydroscopic resin is commercially available.

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

The breathable microporous film is commercially available.

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

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

The Micropump both can be the Micropump of external form；The Micropump can also do into or say the embedded micro-fluidic chip The Micropump of the built in version of its internal sample introduction end structure position or the sample introduction end Near-neighbor Structure position.

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

The gold size sensitive membrane is to be sufficiently mixed 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.

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.

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.

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：

This case is led to dual drive coupling operating mode driving liquid stream in the capillary of the ten Five-channels micro-fluidic chip Flowed in road, joint-detection is carried out to 15 kinds of tumor markerses respectively using multi-channel electrochemical analyzer device.

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 dual drive coupling operating mode driving, various tumour marks The corresponding horseradish peroxidase-labeled that will thing antigen molecule is embedded by gold size sensitive membrane on electrode surface in each passage swells Tumor markers specific antibody captures.

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 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, using the miniature ultrasonic transducer units its The low-power launched, the ultrasonic wave of high-frequency band so that without strong hydrophobic micro-fluidic core of surface chemical modification Compatibility between its tube wall of piece internal pipeline and the test object aqueous solution is significantly increased, and this is sample liquid stream by providing One realistic possibility；Meanwhile using its strong absorbability to ultrasonic wave of dimethyl silicone polymer substrate, shorter In distance, it is, in from the terminal to the very short distance of only several centimeters yardsticks the sample introduction end, reach ultrasound The rapid decrement of intensity of wave, thereby causes the difference of the interfacial tension at the both ends of the micro-fluidic chip, the both ends it Between the difference of interfacial tension can cause a kind of driving force, this kind its function of driving force caused by interfacial tension difference It is to drive sample liquid stream to flow to the terminal direction in strong hydrophobic capillary channel originally；And exist simultaneously in structure The Micropump, its function is the interfacial tension between the inwall of the sample solution and its inner passage of micro-fluidic chip Reduced, under the increased precondition of alternate compatibility by the ul-trasonic irradiation, with the mechanicalness of Micropump pumping strength come with Interfacial tension difference its caused driving force between the both ends of ultrasonic wave induction supports mutually, mutually adjustment, It is mutually coupled, the strength that one driving test liquid flows to the terminal direction flowing is pooled in a manner of Collaboration；This is micro- The presence of pump so that its ultrasonic wave emissive porwer of the miniature ultrasonic transducer units can be allowed to appropriate reduction, and this is for detection Situation containing ultrasonic wave sensitive composition in object is particularly suitable for；And by the ultrasonic transducer and its radiated ultrasonic wave In the presence of, it is possible to increase alternate compatibility, interfacial tension is reduced, and it is special caused by it to provide the both ends interfacial tension difference Driving force, then, in this case, its flow resistance in the micro-channel of sample liquid stream is greatly reduced, and correspondingly, this is micro- Its running resistance of pump is greatly reduced, and so, the Micropump just can be molten for the sample to be carried out than relatively low pumping pressure The pumping work of liquid, because mechanical pumping pressure is greatly reduced, therefore, in such a situation, be just not susceptible to because The micro-channel bubbling of the sample introduction end and its near zone, expansion, change caused by sample introduction terminal tool pumping pressure is excessive Between shape, distortion and the region substrate and cover plate the problems such as stripping etc.；The scheme of this case dual drive coupling running and increasing Add for the handling of sample fluid flow action, can allow for pumping using ultrasonic intensity, ultrasonic frequency, Micropump Power, Micropump pumping pressure etc. multiple indexs are to be directed to the flow rate of the flowing, flowing action includes flow forward or temporary Arrhea dynamic or accelerate flowing etc. flowing action progress multi-parameter accurately to manipulate；By the side of this case dual drive coupling running Case, entirely without must the substrate of the dimethyl silicone polymer material its micro-channel etc. relevant surfaces be carried out with any surface chemistry Modification or surface chemical modification, have altogether dispensed with the laborious procedures of the surface chemical modification or surface chemical modification；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 in the literalness naked poly- diformazan Absorption on its inner surface of pipeline of radical siloxane substrate, and then contain its body phase of the dimethyl silicone polymer substrate to the biology The effect of swallowing up of macromolecular；The Reversible binding thing of the antigen, antibody and antigen and antibody is all to belong to described life certainly The type of thing macromolecular；Because described suction-operated and the described effect of swallowing up effectively are contained, therefore, dependence test As a result will be better able to objectively reflect actual conditions；The effect of the low-power, high-frequency band ultrasonic wave, also include facilitating certainly The quick of Reversible binding reaction between antigen, antibody is reached, and this make it that dependence test operation can be with completeer than faster speed Into.

As described above, the presence of the Micropump so that its ultrasonic wave emissive porwer of the miniature ultrasonic transducer units can be permitted Perhaps appropriateness reduces, then, the feature helps to protect its sensitive coating of the working electrode, is allowed to from ultrasound injury.

Based on this case scheme, completely without carrying out coming to the surface for its relevant surfaces of dimethyl silicone polymer substrate Modification or surface chemical modification operation are learned, therefore, this surface chemical modification layer or surface chemical modification layer not need In the presence of, then, its body phase interior polymer molecule of the dimethyl silicone polymer substrate constantly diffuses to the surface, migrates its institute automatically It is caused that the damaging influence of the surface chemical modification layer or surface chemical modification 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, this kind be very cheap and the easily dimethyl silicone polymer material of processing and fabricating Material be just possible in the micro-fluidic chip its prepare, 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, and 11 be infusion tube, and 12 be Micropump；In legend The spring clamp structure is only the legend structure of signal, and actual spring clamp structure is not limited to the legend spring clamp structure；In legend Arrow indicate the micro-fluidic chip 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 micro-fluidic chip, The structure of the micro-fluidic chip includes being bonded to each other the substrate 5 and cover plate 6 of installing together, and the substrate 5 and cover plate 6 are plate The conduit via mould pressing process or etching technics formation is contained in shape thing or tablet, that face towards the cover plate 6 of the substrate 5 Structure, it is bonded to each other the substrate 5 being installed together and has been built into the micro-fluidic core containing pipeline configuration jointly with the cover plate 6 Piece, the locations of structures of the pipeline are located at the juncture area that the substrate 5 is bonded to each other with the cover plate 6, the both ends of the pipeline respectively with The sample introduction end 4 of the micro-fluidic chip and terminal 3 connect, and the sample introduction end 4 is the injection end of the micro-fluidic chip sample solution, should Terminal 3 is the terminal of sample solution flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal 3 and the sample introduction end 4 are located remotely from each other, and the distance between the terminal 3 and the sample introduction end 4 are between 3 centimetres and 10 centimetres, the terminal 3 and terminal liquid pool Link together, the profile of its structure of the terminal liquid pool is in pond shape, pit shape or cup-shaped, and the terminal liquid pool is used to receive via this Pipeline flow to the sample solution of terminal 3, in the pipeline on diverse location in order or backward be equiped with working electrode and To electrode and reference electrode, the order refers to its locations of structures of the reference electrode closer to the position of terminal 3, institute State backward and refer to the reference electrode locations of structures closer to the position of sample introduction end 4, the working electrode is by electric conductivity electricity Pole and the gold size sensitive membrane for having embedded tumor markers antibody being attached on the conductive electrode are formed, the construction of the pipeline Parallel construction is presented, the pipeline in parallel construction is made up of 15 lateral parallel connections, the presentation parallel construction Its appearance profile of the pipeline is similar to the profile of parallel circuit, and the quantity of the working electrode is 15,15 works Make the installation position of electrode respectively in 15 laterals, and, its top layer gold size of 15 working electrodes Tumor markers antibody in sensitive membrane structure is the 15 kinds of tumour marks that can be specifically bound to tumor markers antigen respectively Will 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 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, fixation is attached on a clamping limb at least in and is equiped with miniature ultrasonic transducer units, this example In, the miniature ultrasonic transducer units 8 are fixed on clamping limb 7 by attaching, and, higher-order of oscillation electric signal transmission cable 9, the height Frequency vibration swings one end of electric signal transmission cable 9 and the miniature ultrasonic transducer units 8 link together；Can also be in two clamping limbs 2nd, all fixed installing miniature ultrasonic transducer units respectively on 7, but in general, miniature ultrasonic only is installed on a clamping limb Wave transducer is just enough to deal with provides a function of facilitating the device to disassemble using needs, the elastic clip 1；The miniature ultrasonic Its major function of wave transducer 8 is that the ultrasonic wave launched using it is tried to reduce in the actual sample introduction test of micro-fluidic chip Interfacial tension between sample solution and the inwall of its inner passage of micro-fluidic chip, can be compatible, also, described in utilization The distance between sample introduction end 4 and the terminal 3 and the installation position of miniature ultrasonic transducer units 8 difference and its experienced Ultrasonic intensity on difference, between its interfacial tension of sample introduction end 4 described in induced synthesis and the terminal 3 its interfacial tension Difference, interfacial tension difference between the micro-fluidic chip both ends 3,4 can between the both ends 3,4 of the micro-fluidic chip shape Into pressure gap, the pressure gap can drive sample solution to be flowed to the direction of terminal 3；The miniature ultrasonic transducer units 8 its Function also include the ultrasonic wave launched with it check large biological molecule contained in sample its in the micro-fluidic chip in it Absorption on portion's channel inner surface, and then check the substrate 5 of the dimethyl silicone polymer material its body phase to the large biological molecule The effect of swallowing up；Softness simultaneously has its function of the substrate 5 of the dimethyl silicone polymer material of elasticity including strong to ultrasonic wave with it The property of strong absorption, ultrasonic wave is absorbed strongly, and thereby between micro-fluidic chip terminal 3 to the sample introduction end 4 Limited short distance within realize the rapid decrement of ultrasonic intensity；And Micropump 12, the Micropump 12 connect with the sample introduction end 4 Connect；The function of the Micropump 12 is the interfacial tension between the inwall of the sample solution and its inner passage of micro-fluidic chip Reduced, under the increased precondition of alternate compatibility by the ul-trasonic irradiation, come with the mechanicalness pumping strength of the Micropump 12 Interfacial tension difference its caused driving force between the both ends 3,4 of ultrasonic wave induction is supported mutually, mutually Adjust, be mutually coupled, the power that one driving test liquid flows to the direction of terminal 3 flowing is pooled in a manner of Collaboration Amount, 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.

Arrow in legend indicate the micro-fluidic chip its in actual motion, by both ends 3,4 pressure differentials drive, its The flow direction of sample liquid stream.

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

The Micropump 12 can customize to specialized factory.

Involved elastic clip 1 is commercially available.It is available for the commercially available alternative spring clamp structure and make that this case structure uses Type 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.

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

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

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 (10)

1. ten Five-channel micro flow control chip devices of common tumor markers examination, it is characterised in that the structure bag of the device Include micro-fluidic chip, the structure of the micro-fluidic chip includes 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, be bonded to each other the substrate that is installed together and be built into the miniflow containing pipeline configuration jointly with the cover plate Chip is controlled, the locations of structures of the pipeline is located at the juncture area that the substrate is bonded to each other with the cover plate, the both ends difference of the pipeline It is connected with the sample introduction end of the micro-fluidic chip and terminal, the sample introduction end is the injection end of the micro-fluidic chip sample solution, should Terminal is the terminal of sample solution flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal and the sample introduction end phase Mutually remote, between 3 centimetres and 10 centimetres, the terminal contacts with terminal liquid pool for the distance between the terminal and the sample introduction end Together, the profile of its structure of the terminal liquid pool is in pond shape, pit shape or cup-shaped, and the terminal liquid pool is used to receive via the pipeline stream Move the sample solution of terminal, in the pipeline on diverse location in order or backward be equiped with working electrode and to electrode with And reference electrode, the order refer to that its locations of structures of the reference electrode refers to closer to the terminal location, the backward Be the reference electrode locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and attaching The gold size sensitive membrane for having embedded tumor markers antibody on the conductive electrode is formed, and structure in parallel is presented in the construction of the pipeline Make, the pipeline in parallel construction is made up of 15 lateral parallel connections, it is described present parallel construction the pipeline its Appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is 15, the dress of 15 working electrodes If position is located in 15 laterals respectively, and, its top layer gold size sensitivity membrane structure of 15 working electrodes In tumor markers antibody be the 15 kinds of tumor markers antibody things that can be specifically bound to tumor markers antigen respectively Matter, 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 are 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 is reduced between sample solution and the inwall of its inner passage of micro-fluidic chip Interfacial tension, can be compatible, also, is installed using the sample introduction end and the terminal with the miniature ultrasonic transducer units Difference in the distance between position difference and its ultrasonic intensity experienced, its interface of sample introduction end described in induced synthesis Difference between tension force and the terminal its interfacial tension, the interfacial tension difference between the micro-fluidic chip both ends can be at this Pressure gap is formed between the both ends of micro-fluidic chip, the pressure gap can drive sample solution to the end flow；Should The ultrasonic wave that miniature ultrasonic transducer units its functions also includes being launched with it check large biological molecule contained in sample its Absorption on the micro-fluidic chip its inner passage inner surface, and then check the substrate of the dimethyl silicone polymer material its body With respect to the effect of swallowing up of the large biological molecule；It is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and 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；And Micropump, the Micropump enter with this Sample end connects；The function of the Micropump is the interface between the inwall of the sample solution and its inner passage of micro-fluidic chip Tension force is reduced, under the increased precondition of alternate compatibility by the ul-trasonic irradiation, and strength is pumped with the mechanicalness of the Micropump Support mutually come interfacial tension difference its caused driving force between the both ends of ultrasonic wave induction, adjust mutually Fit, be mutually coupled, the strength that one driving test liquid flows to the terminal direction flowing is pooled in a manner of Collaboration, should Some high hydroscopic resin particles are filled with terminal liquid pool, its opening end of the terminal liquid pool is covered by breathable microporous film.

2. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, it is capillary channel that the pipeline, which includes the lateral,.

3. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, the thermal decomposition conducting polymer is the electric conductivity material formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated Material.

4. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature Be, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the length of the working electrode Degree is between 1 micron to 15000 microns.

5. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.

6. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, the cover plate its material in structure is dimethyl silicone polymer material.

7. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, the Micropump is miniature piezoelectric pump, miniature peristaltic pump or miniature air driven pump.

8. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, described its thickness of cover plate and substrate in structure is between 1.0 millimeters and 5.0 millimeters.

9. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, the structure of the micro fluidic device also includes higher-order of oscillation electric signal generator, the higher-order of oscillation electric signal transmission cable Its other end is connected with the higher-order of oscillation electric signal generator.

10. ten Five-channel micro flow control chip devices of common tumor markers examination according to claim 1, its feature It is, 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 wave transducer is between 100KHz and 12MHz.