CN107694635A - Six kinds of classifiable tumor mark multichannels while detection micro flow control chip device - Google Patents
Six kinds of classifiable tumor mark multichannels while detection micro flow control chip device Download PDFInfo
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Abstract
The present invention relates to a kind of this case to be referred to as six kinds of classifiable tumor mark multichannels detection micro flow control chip device simultaneously, belongs to analysis testing field.The substrate of the multichannel combined detection micro-fluidic chip of six kinds of classifiable tumor marks is made so that dimethyl silicone polymer is PDMS, is had a clear superiority, but there is also serial problem；This case is directed to the serial problem.This case main points are, the selected PDMS with ecosystem surface of substrate, and its neighbor positions attaches installing miniature ultrasonic transducer units in the liquid stream terminal of the micro-fluidic chip, interfacial tension is reduced with ultrasonic wave, the compatibility between correlative phase is thereby significantly increased, strong absorbability using PDMS to ultrasonic wave simultaneously, reach ultrasonic intensity rapid decrement in short distance, and thus form interfacial tension difference at the both ends of the chip, the interfacial tension difference causes the both ends to form pressure differential, the pressure differential drives liquid stream to be flowed along strong hydrophobic pipeline originally to terminal direction.
Description
Technical field
It is referred to as the detection simultaneously of six kinds of classifiable tumor mark multichannels the present invention relates to a kind of this case to be filled with micro-fluidic chip
Put, belong to analysis testing field.
Background technology
Tumor markers (tumor marker, TM) refers in the generation and breeding of tumour, by tumour cell sheet
Either tumour cell is reacted and caused, a kind of material of the presence of reflection tumour and growth by body caused by body, bag
Include protein, hormone, enzyme (isodynamic enzyme) and oncoprotein etc..The tumor markers in blood samples of patients or body fluid is chemically examined, can be
Early detection tumour in cancer screening, and the effect of observe oncotherapy and judge patient's prognosis.Clinically commonly use at present
Tumor markers has：(1) alpha-fetoprotein (AFP) is the mark of the tumours such as primary carcinoma of liver, carcinoma of testis, oophoroma；(2) cancer embryo
Antigen (CEA) is the mark of the tumours such as digestive system tumor, lung cancer, breast cancer；(3) CA125 (CA125) is ovary
The mark of the tumours such as cancer；(4) CA153 (CA153) is the mark of the tumours such as breast cancer；(5) CA19-9
(CA19-9) it is the mark of digestive system tumor；(6) CA724 (CA724) is the mark of the tumours such as stomach cancer, oophoroma
Thing；(7) carbohydrate antigen 242 (CA242) is the mark of digestive system tumor；(8) CA50 (CA50) is digestive system
The mark of the tumours such as tumour, breast cancer, lung cancer；(9) CYFRA21-1 (Cy211) is the mark of the tumours such as non-small cell lung cancer；
(10) neuronspecific enolase (NSE) is the mark of the tumours such as ED-SCLC, neuroendocrine tumor；(11) before
Row gland specific antigen (PSA) is the tumor markers of prostate cancer；(12) human chorionic gonadotrophin (HCG) is that embryo is thin
The mark of the tumours such as born of the same parents' cancer, trophoblastic tumor (suede cancer, vesicular mole)；(13) thyroglobulin (TG) is the mark of thyroid cancer
Will thing；(14) ferritin (SF) is the mark of the tumours such as digestive system tumor, liver cancer, breast cancer, lung cancer；(15) β 2- microballoons
Albumen (β 2-MG) is in patient body fluids such as chronic lymphocytic leukemia, lymthoma, myeloma, lung cancer, thyroid cancer, nasopharyngeal carcinoma
Middle rise；(16) squamous cell antigen (SCC) is the tumor markerses such as cervical carcinoma, lung squamous cancer, the cancer of the esophagus.Clinically detect at present
The tumor markers overwhelming majority be not only present in malignant tumour, exist in benign tumour, even embryonic tissue, normal group
In knitting.Therefore, tumor markers has dynamic chek and multinomial joint inspection more valuable.So for numerous tumor-markers
Thing, clinically how to selectDifferent tumours understands some relatively special tumor markerses, as CA153 often appears in mammary gland
Cancer；CEA often appears in intestinal cancer, stomach cancer；CA19-9 often appears in intestinal cancer, cancer of pancreas；CA125 often appears in oophoroma etc..Face
Bed doctor can be according to the different mark of different tumor examinations.Same tumour or different types of tumour can have a kind of or several
Kind tumor markers is abnormal；Same tumor markers can occur in different tumours.To improve the auxiliary of tumor markers
Which kind of mark can carry out tumor markers joint-detection to diagnostic value as the follow-up visit monitoring index after treatment with determination,
The complementary tumor markers composition best of breed of several sensitivity of reasonable selection, specific performance, carries out joint-detection.In general
The joint-detection of tumor markers can improve the accuracy to diagnosing tumor.
Only with regard to Diagnostic Value of Several Serum Tumor Markers its joint-detection background of related itself general picture or overview for, can be with
Referring to following Chinese invention patent application case：CN200410041175.3、CN200510026780.8、
CN200610040051.2、CN200910064647.X。
Only for the microfluidic chip technology overall general picture of itself, it is first to may refer to famous micro-fluidic expert Lin Ping Cheng
The raw monograph " diagram Microfluid based Lab on a chip " gone out not long ago, the monograph is published via Science Press, the monograph for
Past of microflow control technique, now, and, vision of the future etc. etc., suffer from detail, be deep into the treatise of detail
Discuss.
So, the focal issue that this case that to have a talk below is paid special attention to.
The basic framework of micro-fluidic chip, including be etched with the substrate of small fluid course and fit together therewith
Cover plate, the small fluid course on the substrate, before upper cover plate is assembled, it is apparent on see to be exactly some micro-channels, to wait until
After covering cover plate thereon, just really closure forms the small fluid course, the conduit inner surface of the micro-channel together with
The part cover plate that surround the micro-channel forms described small fluid course together；So, it is clear that after assembling completes
The small fluid course, the major part of its inner surface area is the inner surface area of that micro-channel, in other words, should
The state or property of micro-channel inner surface substantially determine the integrality or property of the small fluid course；Therefore say, this
The inner surface state or inner surface property of micro-channel of the individual structure on substrate are key factors；In principle, it is any can
The material of its solid forms is kept or kept substantially, can be used to make substrate and cover plate, such as, it can act as substrate and lid
The material of piece can be monocrystalline silicon piece, quartz plate, sheet glass, high polymer for example dimethyl silicone polymer, polymethyl methacrylate,
Makrolon etc.；Certainly, the selection of substrate and the selection of cover plate be able to can also be differed with identical；From material consumption, make
From the point of view of difficulty and application popularization prospect etc. etc., not small difference, the especially choosing of that substrate between these materials be present
Material, have a great influence.
In various substrate making materials, dimethyl silicone polymer, i.e. PDMS, comparatively very easily shaping, at this
It is extremely simple that micro-channel is made on the substrate of sample, and the lower cost for material, base is made with the polydimethyl siloxane material
Piece, micro-channel, and the cover plate phase made with the cheap material such as glass or polypropylene or other plastic sheets are being suppressed or etched thereon
Coordinate, be a kind of more satisfactory selection seemingly；Certainly, patch material can also select to use cheap dimethyl silicone polymer
Material：So, this substrate selection is the scheme of polydimethyl siloxane material, and material is extremely cheap, and making is extremely simple, seems
It should be extremely easy to popularize, promote.
But thing is really not so simple.
First, this polydimethyl siloxane material, that is, the material that alphabetical PDMS is referred to of abridging, itself are a kind of strong
Strong hydrophobic material, micro-channel is built on this material, if without being operated for the modified of the micro-channel surface, that
, after overall assembling is completed, that is, after covering cover plate, because the micro-channel its inner surface in structure occupies most liquid
The inner surface of circulation road, then, its strong hydrophobic property of the PDMS micro-channels inner surface, is deciding factor, it can cause
Similar to the aqueous solution the fine liquid stream of polar liquid by becoming very difficult, its flow resistance is big, or even in general is micro-
Pump is all difficult to promote, certainly, if cover plate also selects to use the PDMS material, then, problem is substantially identical, similar；
Therefore, among prior art, modification is modified particular for the micro-channel inner surface on the PDMS material, is necessary behaviour
Make；So, this is pretty troublesome for the modified operation of PDMS micro-channel inner surfacesThat falls nor this problem, is formed tight
Weight technology puzzlement, be another problem：PDMS polymer molecules inside its body phase of this PDMS material substrate have automatic
The characteristic diffuse to the surface, migrated, the spy that this substrate body phase inside PDMS polymer molecules are diffused to the surface, migrated automatically
Property, the state after modification by its inner surface of that micro-channel of surface modifying and decorating will be caused can not to maintain long enough
Time, the holding time for micro-channel its inner surface state after that is surface-modified be substantially only sufficient to complete laboratory internal
The time of test experiments needs；In other words, by surface modification or the PDMS micro-channel inner surfaces of surface modification, it is modified
The surface state that is formed can not be lasting afterwards or after saying modification, but soon automatically tends to or say and become surface modification again
Surface state before, the strong hydrophobic surface state of that script is returned in the shorter time, then, just think, this
The micro-fluidic chip of sample can largely make, mass storage, be widely popularized, and answer is it is obvious that is, impossible.This
Micro-channel on PDMS material, if not doing surface modification, similar to the aqueous solution the fine liquid stream of polar solvent can not pump it is logical
Cross, chip also cannot just use；And if having done surface modification, its state after modifying can not be persistently kept again, or together
Sample can not popularization and application.
So, how to accomplish that substrate can either be made using cheap PDMS material, and the microflute can be released
Road inner surface decorating state can not persistently, chip can not largely make, largely lay in and then be widely popularized and such a make ability
The puzzlement that the numerous professionals in domain are entangled with for a long time, the highly difficult problem that exactly one its obvious technology barrier can not despise.
Be present many year in the highly difficult problem, so far, not yet properly settled.
Second, the PDMS material of non-surface modification, it is stated that, its surface is strongly hydrophobic above, this strong hydrophobic
Material surface and also have another problem, that is, this strong hydrophobic PDMS surfaces can adsorb large biological molecule, and
And these adsorbed large biological molecules can also further depression further on PDMS surfaces, gradually fall into it is gradually deep, until
It is heavy to be trapped within the body phase of PDMS substrates, in fact, this process, partly it is also due to PDMS material body phase interior polymer
Molecule, which has, to be diffused to the surface, caused by travel motion；Such case, it can also be explained from another angle, i.e. continue not
Disconnectedly from inside PDMS body phases to those polymer molecules of its diffusion into the surface, migration, its result moved, be little by little by that
It has been involved in a bit by the large biological molecule of adsorption within the body phase of PDMS substrates, briefly, these adsorbed biologies
Macromolecular is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up the phenomenon of large biological molecule, its institute
Caused by influence, necessarily cause the severe deviations of all kinds of test data of experiment for being related to large biological molecule.
As described above, the problem of PDMS substrates, is, its not only adsorption large biological molecule, and swallow up biological big point
Son, so, as the large biological molecule of experiment test object, its disappearance will not stop because surface saturation is adsorbed, and
It is, it is constantly adsorbed, also constantly swallowed up.
On PDMS substrates, its body phase is constantly swallowed up and tests showing for associated biomolecule macromolecular in related experiment test process
As, it is to say that another kind, which is explained, substantial amounts of Minute pores in PDMS body phases be present, associated biomolecule macromolecular by after adsorption,
Depression enters these Minute pores, and then is swallowed up；However, inventor thinks, those can allow the sky of miniature scale
Qi leel squeezes into the Minute pores therebetween, not equal to saying that they also can directly allow the large biological molecule of relative large scale to enter
Enter, both difference on yardstick are huge, must not make sweeping generalizations.Explanation is bypassed, in any case, as dependence test analysis object
Large biological molecule is adsorbed by PDMS substrate micro-channels inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is person in charge of reception at ceremonies
Phenomenon existing for sight.
, can be from containment PDMS surfaces pair in order to prevent swallow up effect of this PDMS substrate bodies relative to large biological molecule
The absorption of large biological molecule addresses, and method is chemically modified modification aiming at the PDMS material surface, for
For PDMS is the situation of substrate material, modification exactly is chemically modified to the surface of described micro-channel part, by changing
The micro-channel inner surface of modification is learned, its absorption to large biological molecule can be contained, and then avoid large biological molecule
Swallowed up by PDMS substrate body phases；But or that old problem, that is, the chemical modification on PDMS material surface is modified
Surface state afterwards can not persistently be kept, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface, move automatically
The process of shifting, it soon can become that micro-channel inner surface state being modified by surface chemical modification again script strong and dredge
Water and the state of strong adsorption large biological molecule, in other words, no matter how professionals in the field turn from side to side, the PDMS bases
Its micro-channel inner surface of piece is always rapidly to strong hydrophobic surface state evolution.
So, how can either obtain that PDMS material price is extremely cheap, substrate makes extremely easy benefit, and can is enough
Reach and contain the absorption process of the PDMS substrate micro-channel inner surfaces to large biological molecule for a long time, and then prevent PDMS substrate body phases
The effect of swallowing up to large biological molecule so that related chip manufactured goods be able to maintain that one it is prolonged enough, reasonably guarantee the quality
Phase, it is exactly a very intractable problem.The problem equally makes this area numerous as another problem addressed above
Professional is entangled with, perplexed for a long time, and the problem is equally the highly difficult problem that its obvious technology barrier can not despise.
Also be present many year in the problem, so far, also not yet properly settled.
The content of the invention
The technical problem to be solved by the invention is to provide a package solution, solves totally above
Two problems addressed, also, the solution is applied to build a kind of new can be directed to six kinds than more typical master
Tumor markers is wanted to carry out the micro flow control chip device of examination simultaneously while detection.
The present invention solves the technical problem by following scheme, and the device that the program provides is that a kind of this case is referred to as six kinds
Classifiable tumor mark multichannel is detected and is characterized in micro flow control chip device, the program simultaneously, and the structure of the device includes
Multichannel micro-fluidic chip, the structure of the micro-fluidic chip include being bonded to each other the substrate and cover plate of installing together, the base
Piece and cover plate are plate object or tablet, and that face towards the cover plate of the substrate is contained via mould pressing process or etching work
Skill formed channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, quarter
The window structure that etching technique or simple drilling technology are formed, is bonded to each other the substrate being installed together and is built jointly with the cover plate
Into the micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline is located at the substrate
The interface zone being bonded to each other with the cover plate, its side of the window is blocked by the cover plate and opposite side opens, the structure of the window
Position is exactly the locations of structures of the liquid pool, and the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure position respectively
Sample introduction end liquid pool and terminal liquid pool, the sample introduction end position of the micro-fluidic chip have one or more sample introduction end liquid
Pond, the sample introduction end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the micro-fluidic chip
Terminal location then have a terminal liquid pool, the terminal refers to its chip during the actual sample introduction test of the micro-fluidic chip
The terminal location of interior liquid flowing, the terminal are located remotely from each other with the sample introduction end, and one end of the pipeline is with being located at the described of sample introduction end
Sample introduction end liquid pool UNICOM, the other end of the pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip,
And sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and reference electricity
Pole, the order refer to its locations of structures of the reference electrode closer to the terminal location, and the backward refers to described
For reference electrode locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached to the conduction
Property electrode on the gold size sensitive membrane for having embedded tumor markers antibody form, parallel construction is presented in the construction of the pipeline, described
It is made up of in the pipeline of parallel construction six lateral parallel connections, its appearance profile of the pipeline of the presentation parallel construction is near
The profile of parallel circuit is similar to, the quantity of the working electrode is six, and the installation position of six working electrodes is located at respectively
In six laterals, and, the tumor markers antibody in its top layer gold size sensitivity membrane structure of six working electrodes
It is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, six kinds of antibody materials difference
It is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen is the antigen of broad sense, described anti-
Body is the antibody of broad sense, and its material of the working electrode is that argent material, gold material, carbon material or thermal decomposition are conductive high
Column, sheet or thread is presented in molecule material, its pattern of the working electrode, and its material of the substrate is dimethyl silicone polymer material
Matter, its surface of the substrate are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not pass through any surface
The surface of the primary form of the material of chemical modification or any surface chemical modification, the structure of the device also include miniature ultrasonic
Wave transducer, and, higher-order of oscillation electric signal transmission cable, one end of the higher-order of oscillation electric signal transmission cable is miniature super with this
Acoustic wave transducer links together, and the miniature ultrasonic transducer units are installed in the cover plate or substrate of the micro-fluidic chip with attaching
The position of the neighbouring terminal；Its major function of the miniature ultrasonic transducer units be in the test of micro-fluidic chip actual sample introduction,
The ultrasonic wave launched using it reduces the interfacial tension between sample solution and the inwall of the pipeline, 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 on the inner surface of pipeline
Absorption, and then check swallow up effect of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule；It is described
Its function of the substrate of dimethyl silicone polymer material includes with cover plate and working electrode and electrode and reference electrode is together built
The micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include with it to ultrasonic wave
The property absorbed strongly, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip terminal between the sample introduction end
Limited short distance within realize the rapid decrement of ultrasonic intensity, is filled with some high hydroscopic resins in the terminal liquid pool
Grain, its opening end of the terminal liquid pool are covered by breathable microporous film.
Only for the word of high hydroscopic resin one art-recognized meanings of itself, come for the professional of chemical field
Say, be known.
The high hydroscopic resin is commercially available.
Only for the word of breathable microporous film one art-recognized meanings of itself, for the professional of technical field of membrane,
It is known.
The breathable microporous film is commercially available.
The tumor markers antibody is the antibody of broad sense, and the antibody of the broad sense refers to possessing antibody function or functionally
Can occur to combine with tumor markers involved by various corresponding clinics and form immune complex or exempt from similar to antibody
The material of the analog of epidemic disease compound；The tumor markers antigen is the antigen of broad sense, and the antigen of the broad sense refers to can
Using corresponding antibodies or the material for being functionally similar to antibody need to differentiate, detect involved by the various clinics of enzyme mark detection
Tumor markers.
The 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, shape, 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 micro fluidic device can also include higher-order of oscillation electric signal generator；The higher-order of oscillation electric signal passes
Its other end of transmission cable 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 2 milliwatts and 2000 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 distance between the terminal and the sample introduction end can be the distances of any setting；But the terminal and institute
It is between 3 centimetres and 10 centimetres to state the preferred distance between sample introduction end.
Described its thickness of cover plate and substrate can allow be any setting the thickness for being easy to assembling, the thickness of recommendation or say
Preferable thickness is between 1.0 millimeters and 5.0 millimeters.Less thickness is advantageous to save material.
The application method of this case micro-fluidic chip：
Itd is proposed first based on this case and the first public new liquid stream driving principle, its application of this case micro-fluidic chip are transported
Among work, the new liquid stream driving method is determined completely without involving any additional Micropump.
The interfacial tension difference that this case is formed between micro-fluidic chip both ends caused by the ultrasonic wave,
Driving liquid stream flows in the capillary channel of the six-channel microfluidic chip, right respectively using multi-channel electrochemical analyzer device
Six kinds of classifiable tumor mark antigens carry out joint-detection.
The specific detection of this case micro-fluidic chip is as follows using step：
1st, blood serum sample liquid is added in micro-pipe road, under ultrasonic wave driving, various tumor markers antigen molecules
The tumor markers antibody of the corresponding horseradish peroxidase-labeled embedded by gold size sensitive membrane on electrode surface in each passage
Capture.
2nd, the tumor markers antibody of horseradish peroxidase-labeled is formed with the tumor markers antigen in blood serum sample
Immune complex.
3rd, using multi-channel electrochemical analyzer, the electron mediators such as catechol are added, it is above-mentioned using amperometric detection
Curent change caused by reaction, it is derived from the species and content of various analytes.
4th, result is subjected to comprehensive analysis, comprehensive diagnos is carried out to tumor markers antigen.
It is an advantage of the invention that its close position installs miniature ultrasonic with attaching in the terminal of the micro-fluidic chip
Wave transducer, it is launched using the miniature ultrasonic transducer units low-power, the ultrasonic wave of high-frequency band so that without table
Compatibility between strong hydrophobic its tube wall of micro-fluidic chip internal pipeline and the test object aqueous solution of face chemical modification
It is significantly increased, this is sample liquid stream by providing a realistic possibility；Meanwhile using dimethyl silicone polymer substrate its
To the strong absorbability of ultrasonic wave, in shorter distance, it is, from the terminal to the sample introduction end only
In the very short distance of several centimeters of yardsticks, reach the rapid decrement of ultrasonic intensity, thereby described the two of the micro-fluidic chip
The difference of the interfacial tension is caused at end, and then, utilize its both ends for being formed of the difference of the interfacial tension between the both ends
Between pressure gap, driving sample liquid stream is in the strong hydrophobic capillary channel of such a script to the terminal direction
Flowing.By this case liquid stream drive scheme, entirely without must enter to the substrate and its internal pipeline of the dimethyl silicone polymer material
Any surface chemical modification of row or chemical modification, have altogether dispensed with the laborious procedures of the surface chemical modification or chemical modification；
And the equipment of traditional Micropump etc is altogether dispensed with；On the other hand, the ultrasonic wave of the low-power, high-frequency band,
It can also contain large biological molecule in sample on literalness naked its inner surface of pipeline of dimethyl silicone polymer substrate
Absorption, and then contain the swallow up effect of its body phase of dimethyl silicone polymer substrate to the large biological molecule；The antigen,
The Reversible binding thing of antibody and antigen and antibody is all the type for belonging to described large biological molecule certainly；Due to described suction
Attached effect and the described effect of swallowing up effectively are contained that therefore, dependence test result will be better able to objectively reflect reality
Border situation；The effect of the low-power, high-frequency band ultrasonic wave, also include facilitating the Reversible binding between antigen, antibody anti-certainly
That answers quickly reaches, and 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.
The highly integrated chip structure feature of this case, determine among its application to the requirement of the joint-detection serum
Smaller, this contributes to the body and mind damage for significantly lowering related subject.
Brief description of the drawings
Fig. 1 is its rough outside side view of this case micro flow control chip device.
In figure, 1 is the substrate of dimethyl silicone polymer material, and 2 be cover plate, and 3 be higher-order of oscillation electric signal transmission cable, 4
It is miniature ultrasonic transducer units, 5 be the sample introduction end of the micro-fluidic chip, and 6 be the terminal of the micro-fluidic chip；Legend
In arrow indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, the flowing of its sample liquid stream
Direction.
Embodiment
In this case that Fig. 1 is shown embodiment, the example is characterized in, 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 1 and cover plate 2 of installing together, the substrate 1 and cover plate 2
It is plate object or tablet, that face towards the cover plate 2 of the substrate 1 is contained to be formed via mould pressing process or etching technics
Channel structure, the substrate 1 also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics
Or the window structure that simple drilling technology is formed, it is bonded to each other the substrate 1 being installed together and is built into jointly with the cover plate 2
Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline be located at the substrate 1 with
The interface zone that the cover plate 2 is bonded to each other, its side of the window is blocked by the cover plate 2 and opposite side opens, the structure of the window
Position is exactly the locations of structures of the liquid pool, and the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure position respectively
There is the one or more sample introduction end sample introduction end liquid pool and terminal liquid pool, the position of sample introduction end 5 of the micro-fluidic chip
Liquid pool, the sample introduction end 5 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the micro-fluidic core
Then there is a terminal liquid pool position of terminal 6 of piece, and the terminal 6 refers to during the actual sample introduction test of the micro-fluidic chip it
The terminal location that liquid flows in chip, the terminal 6 are located remotely from each other with the sample introduction end 5, and one end of the pipeline is with being located at sample introduction end 5
The sample introduction end liquid pool UNICOM, the other end of the pipeline with positioned at the micro-fluidic chip the terminal 6 the terminal liquid
Pond UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location 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 position of terminal 6, the backward
Be the reference electrode locations of structures closer to the position of sample introduction end 5, the working electrode is by conductive electrode and patch
The gold size sensitive membrane for having embedded tumor markers antibody being attached on the conductive electrode is formed, and parallel connection is presented in the construction of the pipeline
Construction, the pipeline in parallel construction is made up of six lateral parallel connections, the pipeline of the presentation parallel construction its
Appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is six, the installing position of six working electrodes
Put respectively in six laterals, and, the tumour in its top layer gold size sensitivity membrane structure of six working electrodes
Mark antibody is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, and this six kinds anti-
Body material is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3 respectively, and the antigen is the anti-of broad sense
Original, the antibody are the antibody of broad sense, and its material of the working electrode is argent material, gold material, carbon material or heat
Conducting polymer material is decomposed, column, sheet or thread is presented in the working electrode its pattern, and its material of the substrate 1 is poly- two
Methylsiloxane material, its surface of substrate 1 are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not have
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 is also
Including miniature ultrasonic transducer units 4, and, higher-order of oscillation electric signal transmission cable 3, the higher-order of oscillation electric signal transmission cable 3
One end linked together with the miniature ultrasonic transducer units 4, the miniature ultrasonic transducer units 4 attach to be installed in this micro-fluidic
The position of the cover plate 2 of chip or the neighbouring terminal 6 of substrate 1；Its major function of the miniature ultrasonic transducer units 4 is in miniflow
When controlling the test of chip actual sample introduction, the ultrasonic wave launched using it is reduced between sample solution and the inwall of the pipeline
Interfacial tension, can be compatible, also, utilizes the sample introduction end 5 and the terminal 6 and the miniature ultrasonic transducer units 4
Difference in the distance between installation position difference and its ultrasonic intensity experienced, sample introduction end 5 described in induced synthesis
Difference between its interfacial tension and the terminal 6 its interfacial tension, the interfacial tension between the micro-fluidic chip both ends 5,6
Difference can form pressure gap between the both ends 5,6 of the micro-fluidic chip, and the pressure gap can drive sample solution to institute
Terminal 6 is stated to flow；The ultrasonic wave that its function of the miniature ultrasonic transducer units 4 also includes being launched with it is checked contained in sample
Large biological molecule its absorption on the inner surface of pipeline, and then check the dimethyl silicone polymer material substrate 1 its
Swallow up effect of the body phase to the large biological molecule；The substrate 1 of the dimethyl silicone polymer material its function include with cover plate 2 and
Working electrode and the micro-fluidic chip is together built to electrode and reference electrode, soft dimethyl silicone polymer for simultaneously having elasticity
Its function of the substrate 1 of material is also included with its property to the strong absorption of ultrasonic wave, and ultrasonic wave is absorbed strongly, and thereby
The fast express delivery of ultrasonic intensity is realized within micro-fluidic chip terminal 6 to the limited short distance between the sample introduction end 5
Subtract, is filled with some high hydroscopic resin particles in 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 pressure at two ends difference drive, its try
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 and multiple tracks electrochemical workstation.
Involved miniature ultrasonic transducer units 4 are commercially available；It can also be customized to ultrasonic transducer producer.
Involved higher-order of oscillation electric signal transmission cable 3 is commercially available；Can also to ultrasonic transducer producer customize or to
Cable specialized factory customizes.
Involved higher-order of oscillation electric signal generator market has the product close to needs commercially available；Can also be to relevant speciality producer
Customization.
The multi-channel electrochemical work station can be with commercially available；The multi-channel electrochemical work station can also be according to tool
Body needs whereabouts specialised manufacturers to customize.
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.
Capillary on usual expression and significance refers to hydrophilic capillary glass tube；Capillary channel involved by this case is
The pipeline of hydrophobic capillary form.
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. six kinds of classifiable tumor mark multichannels are detected with micro flow control chip device simultaneously, it is characterised in that the knot of the device
Structure includes multichannel micro-fluidic chip, and 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 are plate object or tablet, that face towards the cover plate of the substrate contain via mould pressing process or
Etching technics formed channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via stamper
The window structure that skill, etching technics or simple drilling technology are formed, is bonded to each other the substrate being installed together and is total to the cover plate
With the micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto has been built into, the locations of structures of the pipeline is located at
The interface zone that the substrate is bonded to each other with the cover plate, its side of the window is blocked by the cover plate and opposite side opens, the window
Locations of structures be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are to be located at different structure respectively
The sample introduction end liquid pool and terminal liquid pool of position, the sample introduction end position of the micro-fluidic chip have it is one or more it is described enter
Sample end liquid pool, the sample introduction end refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the miniflow
The terminal location of control chip then has a terminal liquid pool, when the terminal refers to that the actual sample introduction of the micro-fluidic chip is tested
The terminal location that liquid flows in its chip, the terminal are located remotely from each other with the sample introduction end, and one end of the pipeline is with being located at sample introduction end
The sample introduction end liquid pool UNICOM, the other end of the pipeline with positioned at the micro-fluidic chip the terminal the terminal liquid pool
UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and
Reference electrode, the order refer to its locations of structures of the reference electrode closer to the terminal location, what the backward referred to
Be the reference electrode locations of structures closer to the sample introduction end position, the working electrode is by conductive electrode and is attached to
The gold size sensitive membrane for having embedded tumor markers antibody on the conductive electrode is formed, and structure in parallel is presented in the construction of the pipeline
Make, the pipeline in parallel construction is made up of six lateral parallel connections, and the pipeline that parallel construction is presented is outside it
Shape profile is similar to the profile of parallel circuit, and the quantity of the working electrode is six, the installation position of six working electrodes
Respectively in six laterals, and, the tumour mark in its top layer gold size sensitivity membrane structure of six working electrodes
Will thing antibody is the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen respectively, six kinds of antibody
Material is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3 respectively, and the antigen is the anti-of broad sense
Original, the antibody are the antibody of broad sense, and its material of the working electrode is argent material, gold material, carbon material or heat
Conducting polymer material is decomposed, column, sheet or thread is presented in its pattern of the working electrode, and its material of the substrate is poly- diformazan
Radical siloxane material, its surface of the substrate are the surfaces of primary form, the surface of the primary form its be intended to refer to and do not pass through
The surface of the primary form of the material of any surface chemical modification or any surface chemical modification is crossed, the structure of the device is also wrapped
Miniature ultrasonic transducer units are included, and, higher-order of oscillation electric signal transmission cable, one end of the higher-order of oscillation electric signal transmission cable
Linked together with the miniature ultrasonic transducer units, the miniature ultrasonic transducer units are installed in the lid of the micro-fluidic chip with attaching
The position of the neighbouring terminal of piece or substrate；Its major function of the miniature ultrasonic transducer units 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 and the terminal and the miniature ultrasonic transducer units installation position
Difference in distance difference and its ultrasonic intensity experienced, its interfacial tension of sample introduction end described in induced synthesis with it is described
Difference between its interfacial tension of terminal, the interfacial tension difference between the micro-fluidic chip both ends can be in the micro-fluidic chip
The both ends between form pressure gap, the pressure gap can drive sample solution to the end flow；The miniature ultrasonic
The ultrasonic wave that transducer its function also includes being launched with it check large biological molecule contained in sample its in the pipeline
Absorption on inner surface, and then check swallow up work of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule
With；Its function of the substrate of the dimethyl silicone polymer material is included with cover plate and working electrode and to electrode and reference electrode one
With building the micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function also include it is right with its
The property that ultrasonic wave absorbs strongly, ultrasonic wave is absorbed strongly, and thereby in micro-fluidic chip terminal to the sample introduction
The rapid decrement of ultrasonic intensity is realized within limited short distance between end, some high water absorptions are filled with the terminal liquid pool
Resin particle, its opening end of the terminal liquid pool are covered by breathable microporous film.
2. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, it is capillary channel that the pipeline, which includes the lateral,.
3. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, the thermal decomposition conducting polymer is the electric conductivity formed by polyimides or polyacrylonitrile after anoxybiotic is heat-treated
Material.
4. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
Be characterised by, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the working electrode
Length between 1 micron to 15000 microns.
5. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.
6. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, the cover plate its material in structure is dimethyl silicone polymer material.
7. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, the distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.
8. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, described its thickness of cover plate and substrate in structure is between 1.0 millimeters and 5.0 millimeters.
9. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, the structure of the micro fluidic device also includes higher-order of oscillation electric signal generator, the higher-order of oscillation electric signal transmission
Its other end of cable is connected with the higher-order of oscillation electric signal generator.
10. six kinds of classifiable tumor mark multichannels according to claim 1 detect with micro flow control chip device simultaneously, its
It is characterised by, for its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units between 2 milliwatts and 2000 milliwatts, this is miniature
The frequency of its ultrasonic wave operationally launched of ultrasonic transducer is between 100KHz and 12MHz.
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| CN110208534A (en) * | 2019-05-27 | 2019-09-06 | 上海理工大学 | Self-priming Diagnostic Value of Several Serum Tumor Markers Multi-example detection chip |
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| CN110208534A (en) * | 2019-05-27 | 2019-09-06 | 上海理工大学 | Self-priming Diagnostic Value of Several Serum Tumor Markers Multi-example detection chip |
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