CN107225002A - The classifiable tumor markers in detecting micro flow control chip device that women is applicable - Google Patents
The classifiable tumor markers in detecting micro flow control chip device that women is applicable Download PDFInfo
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Abstract
The present invention relates to the classifiable tumor markers in detecting micro flow control chip device that a kind of women is applicable, belong to analysis testing field.The substrate for the classifiable tumor markers in detecting micro-fluidic chip that women is applicable is made so that PDMS is dimethyl silicone polymer, has price and technologic advantage, but also have any problem；This case is intended to overcome the difficulty.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 test liquid stream terminal of the micro-fluidic chip, simultaneously, ultrasonic wave damper is installed at the sample introduction end of the micro-fluidic chip, reach ultrasonic intensity rapid decrement in short distance, and thus form interfacial tension difference at the two ends of the chip, the interfacial tension difference causes the two ends to form pressure differential, and the pressure differential drives test liquid stream to be flowed along strong hydrophobic capillary level passage originally to terminal direction.
Description
Technical field
The present invention relates to the classifiable tumor markers in detecting micro flow control chip device that a kind of women is applicable, belong to analysis test
Field.
Background technology
Tumor markers (tumor marker, TM) refers in the generation and breeding of tumour, is produced in itself by tumour cell
Raw either reacted by body tumour cell and produce, reflection tumour exist and growth a class material, including protein,
Hormone, enzyme (isodynamic enzyme) and oncoprotein etc..The tumor markers in blood samples of patients or body fluid is chemically examined, can be in cancer screening
Early detection tumour, and observe the curative effect of oncotherapy and judge patient's prognosis.The tumor markers clinically commonly used at present has：
(1) alpha-fetoprotein (AFP) is the mark of the tumours such as primary carcinoma of liver, carcinoma of testis, oophoroma；(2) carcinomebryonic antigen (CEA) is digestion
The mark of the tumours such as system tumor, lung cancer, breast cancer；(3) CA125 (CA125) is the mark of the tumours such as oophoroma；
(4) CA153 (CA153) is the mark of the tumours such as breast cancer；(5) CA19-9 (CA19-9) is digestive system tumor
Mark；(6) CA724 (CA724) is the mark of the tumours such as stomach cancer, oophoroma；(7) carbohydrate antigen 242 (CA242)
For the mark of digestive system tumor；(8) CA50 (CA50) is the mark of the tumours such as digestive system tumor, breast cancer, lung cancer
Thing；(9) CYFRA21-1 (Cy211) is the mark of the tumours such as non-small cell lung cancer；(10) neuronspecific enolase (NSE)
For the mark of the tumours such as ED-SCLC, neuroendocrine tumor；(11) PSA (PSA) is prostate cancer
Tumor markers；(12) human chorionic gonadotrophin (HCG) is that embryonic cell carcinoma, trophoblastic tumor (suede cancer, vesicular mole) etc. are swollen
The mark of knurl；(13) thyroglobulin (TG) is the mark of thyroid cancer；(14) ferritin (SF) is digestive system tumor, liver
The mark of the tumours such as cancer, breast cancer, lung cancer；(15) B2M (β 2-MG) is in chronic lymphocytic leukemia, lymph
Raised in the patient body fluids such as knurl, myeloma, lung cancer, thyroid cancer, nasopharyngeal carcinoma；(16) squamous cell antigen (SCC) be cervical carcinoma,
The tumor markerses such as lung squamous cancer, the cancer of the esophagus.The tumor markers overwhelming majority clinically detected at present is not only present in malignant tumour
In, exist in benign tumour, embryonic tissue, even in normal structure.Therefore, tumor markers has dynamic chek and multinomial
Joint inspection is more valuable.So for numerous tumor markerses, clinically how to selectDifferent tumours understands some phases
To special tumor markers, such as CA153 often appears in breast cancer；CEA often appears in intestinal cancer, stomach cancer；CA19-9 is often appeared in
Intestinal cancer, cancer of pancreas；CA125 often appears in oophoroma etc..Clinician can be according to the different mark of different tumor examinations.
Same tumour or different types of tumour can have one or more of tumor markerses abnormal；Same tumor markers can be in difference
Tumour in occur.To improve the additive diagnostic value of tumor markers and determining which kind of mark can be supervised as the follow-up after treatment
Index is surveyed, tumor markers joint-detection, the complementary tumor markers group of several sensitivity of reasonable selection, specific performance can be carried out
Into best of breed, joint-detection is carried out.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, may refer to
Lower Chinese invention patent application case：CN200410041175.3、CN200510026780.8、CN200610040051.2、
CN200910064647.X。
Only for the microfluidic chip technology overall general picture of itself, famous micro-fluidic expert Mr. Lin Ping Cheng may refer to soon
Before the monograph " diagram Microfluid based Lab on a chip " that goes out, the monograph is published via Science Press, and the monograph is for micro-fluidic
Past of technology, now, and, in terms of vision of the future etc., suffer from detail, be deep into long discussion of detail.
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 it is etched with the substrate of small fluid course and the cover plate fit together therewith,
Small fluid course on the substrate, assembling upper cover plate before, it is apparent on see to be exactly some micro-channels, wait until thereon
After covering cover plate, just real closure forms the small fluid course, and the conduit inner surface of the micro-channel is together with surround this
The part cover plate of micro-channel constitutes described small fluid course together；So, it is clear that this after assembling is completed is small
Fluid course, the major part of its inner surface area is the inner surface area of that micro-channel, in other words, in the micro-channel
The state or property on surface substantially determine the integrality or property of the small fluid course；Therefore say, this is built in base
The inner surface state or inner surface property of micro-channel on piece are key factors；In principle, it is any to keep or protecting substantially
The material of its solid forms is held, can be used to make substrate and cover plate, such as, the material that can act as substrate and cover plate can be with
It is monocrystalline silicon piece, quartz plate, sheet glass, high polymer such as dimethyl silicone polymer, polymethyl methacrylate, makrolon etc.
Deng；Certainly, the selection of substrate and the selection of cover plate be able to can also be differed with identical；From material consumption, manufacture difficulty and
From the point of view of in terms of application popularization prospect etc., between these materials exist not small difference, the especially selection of that substrate, influence compared with
Greatly.
In various substrate making materials, dimethyl silicone polymer, i.e. PDMS, comparatively very easily shaping, so
Substrate on make that micro-channel is extremely simple, and the lower cost for material makes substrate with the polydimethyl siloxane material,
Micro-channel is being suppressed or etched thereon, and is being engaged with the cover plate that the cheap material such as glass or polypropylene or other plastic sheets makes,
It is a kind of more satisfactory selection seemingly；Certainly, patch material can also select to use cheap polydimethyl siloxane material：
So, this substrate selection is the scheme of polydimethyl siloxane material, and material is extremely cheap, makes extremely simple, seems and should also be as
It is extremely easy to popularize, promotes.
But, thing is really not so simple.
First, this polydimethyl siloxane material, that is, the material that the letter PDMS that abridges is referred to, itself are a kind of strong
Hydrophobic material, micro-channel is built on this material, if without being operated for the modified of the micro-channel surface, then,
After overall assembling is completed, that is, cover after cover plate, because the micro-channel its inner surface in structure occupies most liquid circulation
The inner surface in road, then, its strong hydrophobic property of the PDMS micro-channels inner surface, is deciding factor, it can cause class
Be similar to the aqueous solution the fine liquid stream of polar liquid by becoming very difficult, its flow resistance is big, or even general Micropump is all
It is 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
Operation；So, this is pretty troublesome for the modified operation of PDMS micro-channel inner surfacesThat falls nor this problem, structure
Perplex into serious technical, be another problem：PDMS polymer molecules tool inside its body phase of this PDMS material substrate
There are PDMS polymer molecules inside the characteristic for diffusing to the surface, migrating automatically, this substrate body phase to diffuse to the surface, move automatically
The characteristic of shifting, will cause the state after modification by its inner surface of that micro-channel of surface modifying and decorating can not maintain foot
Enough long time, the holding time for micro-channel its inner surface state after that is surface-modified is substantially only sufficient to complete in laboratory
The time of portion's test experiments needs；In other words, by surface modification or the PDMS micro-channel inner surfaces of surface modification, its
The surface state that is formed can not be lasting after modification or after saying modification, but soon automatically tend to or say and become surface again and change
Surface state before property, returns to the strong hydrophobic surface state of that script in the shorter time, then, just think,
Such micro-fluidic chip can largely make, mass storage, be widely popularized, and answer is it is obvious that is, impossible.
Micro-channel on this PDMS material, can not pump similar to the fine liquid stream of polar solvent of the aqueous solution if not doing surface modification
Send and pass through, chip also cannot just be used；And if having done surface modification, the state after its modification can not be persistently kept again, also
Being equally can not popularization and application.
So, how to accomplish that substrate can either be made using cheap PDMS material, and table in the micro-channel can be released
Face decorating state can not persistently, chip can not largely make, largely lay in so that be widely popularized it is such a make this area it is numerous specially
The puzzlement that industry personnel 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, above it is stated that, its surface is strongly hydrophobic, this strong hydrophobic
Material surface and also 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 gradually deep, directly
It is trapped in heavy within the body phase of PDMS substrates, in fact, this process is partly also due to inside PDMS material body phase
Polymer molecule, which has, to be diffused to the surface, caused by travel motion；Such case, can also be explained from another angle, i.e.
Continuously from inside PDMS body phases to its diffusion into the surface, migration those polymer molecules, its result moved, be by
Gradually those are involved within the body phase of PDMS substrates by the large biological molecule of adsorption, briefly, these are inhaled
Attached large biological molecule is exactly to be swallowed up by PDMS substrate body phases；So, this PDMS substrates body phase swallows up large biological molecule
Phenomenon, the influence caused by it necessarily causes the severe deviations of all kinds of test data of experiment for being related to large biological molecule.
As described above, be the problem of PDMS substrates, its not only adsorption large biological molecule, and swallow up large biological molecule,
So, as the large biological molecule of experiment test object, its disappearance will not stop because the absorption of surface saturation, but, no
It is disconnected adsorbed, also constantly swallowed up.
On PDMS substrates in related experiment test process its body phase constantly swallow up test associated biomolecule macromolecular phenomenon, separately
It is to say that one kind, which is explained, there are substantial amounts of Minute pores in PDMS body phases, and associated biomolecule macromolecular is by after adsorption, depression
Into these Minute pores, and then swallowed up；However, inventor thinks, those can allow the air point of miniature scale
Son squeezes into the Minute pores therebetween, and being not equal to them also can directly allow that the large biological molecule of relative large scale enters, and two
Person's difference on yardstick is huge, must not make sweeping generalizations.Explanation is bypassed, in any case, the biology of object is analyzed as dependence test
Macromolecular is adsorbed by PDMS substrate micro-channels inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is known objective
The phenomenon of presence.
, can be from containment PDMS surfaces to life in order to prevent this PDMS substrate bodies relative to the effect of swallowing up of large biological molecule
The absorption of thing macromolecular is addressed, 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 quilt
PDMS substrate body phases are swallowed up；But, or that old problem, that is, the chemical modification on PDMS material surface changes
Surface state after property can not persistently be kept, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface automatically,
The process of migration, 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
Its micro-channel inner surface of substrate 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, can reach again
Growth stage contains absorption process of the PDMS substrate micro-channel inner surfaces to large biological molecule, and then prevents PDMS substrate body phases
The effect of swallowing up to large biological molecule so that related chip manufactured goods are able to maintain that a prolonged enough, rational shelf-life,
It is exactly a very intractable problem.The problem makes the numerous specialties in this area as another problem addressed above, equally
Personnel are entangled with, perplexed for a long time, and the problem is equally the highly difficult problem that its obvious technology barrier can not despise.The hardly possible
Also be present many year in topic, so far, also not yet properly settled.
The content of the invention
The technical problems to be solved by the invention are to provide a package solution, solve what is addressed above totally
Two problems, also, the solution is applied to build a kind of new can especially need concern for belonging to women physical examination
Seven kinds than more typical primary tumor mark carry out simultaneously examination, while detection micro flow control chip device.
The present invention solves the technical problem by following scheme, and the device that the program is provided is that the typical case that a kind of women is applicable swells
Tumor markers joint-detection micro flow control chip device, it is characterised in that the structure of the device includes multichannel micro-fluidic chip,
The structure of the micro-fluidic chip includes installing substrate and cover plate together bonded to each other, the substrate and cover plate be plate object or
Contain the channel structure via mould pressing process or etching technics formation, the base in tablet, that face towards the cover plate of the substrate
Piece also contains and is connected and pierces being formed via mould pressing process, etching technics or simple drilling technology for the substrate with the channel structure
Window structure, substrate bonded to each other being installed together and the cover plate have been built into containing pipeline configuration and phase therewith jointly
The micro-fluidic chip of liquid pool structure even, the locations of structures of the pipeline is located at the substrate and cover plate interface zone bonded to each other,
Its side of the window is blocked by the cover plate and opposite side is opened, and the locations of structures of the window is exactly the locations of structures of the liquid pool, institute
Stating liquid pool has two kinds, and two kinds of liquid pools are the sample introduction end liquid pool and terminal liquid pool positioned at different structure position respectively, and this is micro-fluidic
The sample introduction end position of chip has one or more sample introduction end liquid pool, and the sample introduction end refers to that the micro-fluidic chip is real
The injection end position of detected solution during the sample introduction of border, then has a terminal liquid pool in the terminal location of the micro-fluidic chip, described
Terminal refers to the terminal location of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested, the terminal and the sample introduction end
It is located remotely from each other, one end of the pipeline and the sample introduction end liquid pool UNICOM positioned at sample introduction end, the other end of the pipeline are micro- with being located at this
The terminal liquid pool UNICOM of the terminal of fluidic chip, and, sequentially or backward be respectively installed in the pipeline it is different
Working electrode on position and to electrode and reference electrode, the order refers to that its locations of structures of the reference electrode is more leaned on
The nearly terminal location, the backward refers to the reference electrode locations of structures closer to the sample introduction end position, the work
Electrode is made up of conductive electrode and the gold size sensitive membrane for having embedded tumor markers antibody being attached on the conductive electrode,
Parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is made up of eight lateral parallel connections, and the presentation is simultaneously
Its appearance profile of the pipeline of connection construction is similar to the profile of parallel circuit, and the quantity of the working electrode is eight, this eight
The installation position of working electrode is located in eight laterals respectively, and, its top layer gold size of eight working electrodes is sensitive
Tumor markers antibody in membrane structure is the eight kinds of tumor markers antibody that can be specifically bound to tumor markers antigen respectively
Material, eight kinds of antibody materials be respectively tumor markers antibody CA199, CA153, CA125, CA242, AFP, CEA,
EB and β-HCG, the antigen is the antigen of broad sense, and the antibody is the antibody of broad sense, and its material of the working electrode is gold
The silver-colored material of category, gold material, carbon material or thermal decomposition conducting polymer material, the working electrode its pattern presentation column,
Sheet is thread, and its material of the substrate is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, the original
The surface of raw form its be intended to refer to not by any surface chemical modification or the original of the material of any surface chemical modification
The surface of raw form, the structure of the device also includes miniature ultrasonic transducer units, and, higher-order of oscillation electric signal transmission cable,
One end of the higher-order of oscillation electric signal transmission cable links together with the miniature ultrasonic transducer units, the miniature ultrasonic transducer units
It is installed in the position of the cover plate of the micro-fluidic chip or the neighbouring terminal of substrate with attaching；Its master of the miniature ultrasonic transducer units
It is that in the test of micro-fluidic chip actual sample introduction, the ultrasonic wave launched using it reduces sample solution and the pipeline to want function
Inwall between interfacial tension, can be compatible, also, utilize the sample introduction end and the terminal and the miniature ultrasonic
Difference in the distance between wave transducer installation position difference and its ultrasonic intensity experienced, enters described in induced synthesis
Interfacial tension between difference between its interfacial tension of sample end and the terminal its interfacial tension, the micro-fluidic chip two ends is poor
Different to form pressure gap between the two ends of the micro-fluidic chip, the pressure gap can drive sample solution to the terminal stream
It is dynamic；Its function of the miniature ultrasonic transducer units also includes checking large biological molecule contained in sample with its ultrasonic wave launched
Its absorption on the inner surface of pipeline, and then check its body phase of the substrate of the dimethyl silicone polymer material to the biology greatly point
The effect of swallowing up of son；Its function of the substrate of the dimethyl silicone polymer material is included with cover plate and working electrode and to electrode and ginseng
Together build the micro-fluidic chip than electrode, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include
With its property to the strong absorption of ultrasonic wave, ultrasonic wave is absorbed strongly, and thereby the micro-fluidic chip terminal to should
The rapid decrement of ultrasonic intensity is realized within limited short distance between sample introduction end；And, ultrasonic wave damper, the ultrasound
Its profile of ripple damper is in block, tabular or bar-shaped；Its material of the ultrasonic wave damper is microcellular rubber, sponge rubber or poly- ammonia
Ester micro-pore elastomer；And, elastic clip, the elastic force accompanies two clamping limbs, and the medial surface of each clamping limb is clamped with another
The medial surface of arm is relative, and the medial surface of one of clamping limb is close to the sample introduction end of the micro-fluidic chip, another clamping limb
Medial surface is close to the ultrasonic wave damper, and the elastic clip is with its gripping strength by the sample introduction end of the micro-fluidic chip and the ultrasonic wave
Damper is clipped together bonded to each otherly, the strong consumption to ultrasonic wave that the ultrasonic wave damper is brought with its material,
Further support to realize ultrasonic intensity within micro-fluidic chip terminal to the limited short distance between the sample introduction end
Rapid decrement, and thereby further expand the difference between described its interfacial tension of sample introduction end and the terminal its interfacial tension.
The tumor markers antibody is the antibody of broad sense, and the antibody of the broad sense refers to possessing antibody function or functionally similar
Can occur to combine and formed immune complex or immune compound with the various involved tumor markerses of corresponding clinic in antibody
The material of the analog of thing；The tumor markers antigen is the antigen of broad sense, and the antigen of the broad sense is referred to can be using accordingly
Antibody or be functionally similar to antibody material carry out enzyme mark detection it is various it is clinical involved the need for differentiate, the tumor-marker of detection
Thing.
The gold size sensitive membrane is to be sufficiently mixed chitosan gold size solution and tumor markers antibody-solutions uniformly, uses point sample instrument
Point sample is coated on specified structure position, and forms its drying and forming-film.Tumor markers antibody in the gold size sensitive membrane is equal
The tumor markers antibody marked for horseradish peroxidase or glucose oxidase, the gold size sensitive membrane has been included as fixing
Above-mentioned each tumor markers antibody and introduce complementary medium therein, the complementary medium for example chitosan, cellulose acetate,
Gelatin is 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 arbitrarily selected
Size, still, for less with the consideration in terms of prepare liquid sample and reduction reagent loss, the pipeline includes described as far as possible
The passage of the preferred capillary level of lateral, the passage of the capillary level implies that the interior of its internal diameter and the capillary on ordinary meaning
The suitable passage in footpath.The shape of cross section of its inner passage of capillary can be arbitrary shape, the shape of cross section example
Such as circular, oval, square, rectangle, bar shaped, naturally it is also possible to be arbitrarily the presence of the linear of bending, also, the hair
The interior shape of tubule is with the extension of pipeline, and the shape of cross section of different parts can also allow to be different shapes.Only with regard to hair
For the word of tubule 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 may each be any choosing
Fixed shape, the arbitrarily selected shape such as column, sheet, strip or thread etc..It is described to electrode and the ginseng
Art-recognized meanings than the 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 scrotiform structure for transitional liquid storage
Make, its shape of the inner chamber of each liquid pool may each be arbitrarily selected shape, the cavity shape such as cylindrical empty
Cavity-like, square column type cavity-like, oblong cavity shape or spherical hollow space shape etc..
It is public only for professional of the word of ultrasonic transducer one art-recognized meanings of itself for ultrasonic technology field
Know.
Various sizes, variously-shaped ultrasonic transducer are commercially available；Commercially available miniature ultrasonic transducer units its sizes can be with
It is small 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 itself and
Speech, is known general technology for the professional in ultrasonic technology field.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 technology；Together
Sample, the technology of hole-opening is even more known simple technique on naked PDMS substrates.This case is not also to this expansion superfluous words.
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 transmission electricity
Its other end of 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, for the professional in ultrasonic technology field, be
It is simple and known；The higher-order of oscillation electric signal generator can be customized to ultrasonic instrument specialized factory.
The preferred scope of its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units be between 2 milliwatts and 2000 milliwatts it
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, the annex such as multiple tracks electrochemical workstation etc., institute certainly
The art-recognized meanings for stating multiple tracks electrochemical workstation are known.Each working electrode for being related in this case microfluidic chip structure and
, can be respectively via corresponding special the corresponding interface got lines crossed with the multiple tracks electrochemical workstation to electrode and reference electrode etc.
Coupled.It is described that special to get lines crossed be for each the corresponding interface of each electrode and the multiple tracks electrochemical workstation is carried out into phase
The private cable being mutually coupled with.The micro-fluidic chip in this case device, its structure can also include micro-valve, the number of the micro-valve
Amount is not limited, according to actual needs, and the micro-valve can be installed in any required position installed in the microfluidic chip structure；
The word of micro-valve one is for the professional of micro fluidic chip technical field, and the art-recognized meanings of itself are known；The micro-valve
The manufacturing technology of itself 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, it is however recommended to
Or say preferred its scope of the diameter between 0.1 micron to 2000 microns；The length of the working electrode can permit
Permitted to be that any setting is easily installed the length used, it is however recommended to or to say preferred its scope of the length be at 1 micron
To between 15000 microns.
The gold size for being installed in the working electrode surface layer by spraying or point sample instrument point sample or the coating of other appropriate process is quick
Feel film, its thicknesses of layers can allow be any setting treat sample measuring liquid occur electrical signals response thickness, still, push away
Preferred thickness is between 10 nanometers and 200 nanometers in other words for the thickness recommended.
The cover plate in chip structure, its material can allow to be any electrical insulating property material, for example：Polypropylene, glass
Glass, polymethyl methacrylate, dimethyl silicone polymer, etc., in order to make smaller size of micro-fluidic chip, such as do
Realized into the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and 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
It is used as the cover plate using dimethyl silicone polymer, is also that this case technical scheme is allowed.
The preferred scope of the distance between the terminal and the sample introduction end is between 3 centimetres and 10 centimetres.
Described its thickness of cover plate and substrate can allow be any setting be easy to assembling thickness, the thickness of recommendation or say preferably
Thickness be between 1.0 millimeters and 5.0 millimeters.Less thickness is conducive to saving material.
Described its size of ultrasonic wave damper is not limited, and described its length of ultrasonic wave damper, width, thickness may each be basis
Need the arbitrary size of setting.
It is known in field of rubber technology only for the word of microcellular rubber one art-recognized meanings of itself.
The microcellular rubber is commercially available.
It is known in field of rubber technology only for the word of sponge rubber one art-recognized meanings of itself.
The sponge rubber is commercially available.
It is known in technical field of macromolecules only for the word of microporous polyurethane elastomer one art-recognized meanings of itself.
The polyurethane micropore elastomer material is commercially available.
The microcellular rubber preferred microporous silicon rubber or micropore fluorubber.
It is known in field of rubber technology only for the word of micropore silicon rubber one art-recognized meanings of itself.
The microporous silicon Rubber market is on sale.
It is known in field of rubber technology only for the word of micropore fluorubber one art-recognized meanings of itself.
The micropore fluorubber is commercially available.
The word of elastic clip one art-recognized meanings of itself are known.
The elastic clip is commercially available.
The application method of this case micro-fluidic chip：
Proposed first based on this case and the first public New stream driving principle, its application running of this case micro-fluidic chip
In, the New stream driving method is determined completely without involving any additional Micropump.
The interfacial tension difference of this case to be formed between micro-fluidic chip two ends caused by the ultrasonic wave, drives liquid
Stream flows in the capillary channel of eight channel microfluidic chip, using eight passage electrochemical analytical instruments respectively to eight kinds of typical cases
Markers in diagnosis antigen is detected.
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 are each
In passage on electrode surface gold size sensitive membrane embed corresponding horseradish peroxidase-labeled tumor markers antibody capture.
2nd, the tumor markers antigen in the tumor markers antibody and blood serum sample of horseradish peroxidase-labeled forms immune multiple
Compound.
3rd, using multi-channel electrochemical analyzer, the electron mediators such as catechol are added, using the above-mentioned reaction of amperometric detection
Caused curent change, 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.
Changed it is an advantage of the invention that installing miniature ultrasonic in its close position attaching of the terminal of the micro-fluidic chip
Can device, it is launched using the miniature ultrasonic transducer units low-power, the ultrasonic wave of high-frequency band so that without coming to the surface
The compatibility learned between strong hydrophobic its tube wall of micro-fluidic chip internal pipeline and the test object aqueous solution of modification significantly increases
Plus, this is test liquid stream by there is provided a realistic possibility；Meanwhile, using dimethyl silicone polymer substrate its to ultrasound
The strong absorbability of ripple, in shorter distance, it is, from the terminal to only several centimeters the sample introduction end
In the very short distance of yardstick, reach the rapid decrement of ultrasonic intensity, thereby cause institute at the two ends of the micro-fluidic chip
The difference of interfacial tension is stated, and then, utilize the pressure between its two ends for being formed of the difference of the interfacial tension between the two ends
Difference, driving test liquid stream flows in the strong hydrophobic capillary channel of such a script to the terminal direction.By this
Case liquid stream drive scheme, entirely without any come to the surface must be carried out to the substrate and its internal pipeline of the dimethyl silicone polymer material
Modification or chemical modification are learned, the laborious procedures of the surface chemical modification or chemical modification have been altogether dispensed with；And altogether dispense with biography
The equipment of Micropump in meaning of uniting etc；On the other hand, the ultrasonic wave of the low-power, high-frequency band, additionally it is possible to contain in sample
Absorption of the large biological molecule on literalness naked its inner surface of pipeline of dimethyl silicone polymer substrate, and then contain that this gathers
Swallow up effect of its body phase of dimethyl siloxane piece to the large biological molecule；The antigen, antibody and antigen and antibody
Reversible binding thing is all the type for belonging to described large biological molecule certainly；Due to described suction-operated and the described work that swallows up
With effectively being contained, therefore, dependence test result will be better able to objectively reflect actual conditions；The low-power, height are again and again
The effect of section ultrasonic wave, also includes facilitating what the Reversible binding between antigen, antibody react quickly to reach, this causes correlation certainly
Test operation can be with than the completion of faster speed.
Due to the surface chemical modification for the dimethyl silicone polymer substrate its relevant surfaces or chemical modification behaviour need not be carried out
Make, therefore, this surface chemical modification layer or chemically modified layer not need to exist, then, the dimethyl silicone polymer
Its body phase interior polymer molecule of substrate constantly diffuses to the surface automatically, migrate caused by it to surface chemical modification layer or
The damaging influence of chemically modified layer is also just not present.
This case its installation position is located at the ultrasonic wave damper at the sample introduction end, from its terminal of this case micro-fluidic chip to
Said on the sport technique segment that ultrasonic intensity rapid decrement is realized in the such short distance in sample introduction end, its effect of the ultrasonic wave damper is suitable
In the size expansion device of micro-fluidic chip, in other words, the ultrasonic wave damper its function as and significantly extend miniflow
Its terminal of chip is controlled the distance between to sample introduction end, is the equal of from several centimetres of extension of script to more than ten centimetres of length
Or twenties centimetres of length or longer length, so so that described terminal to the ultrasonic wave between sample introduction end in short distance
The sport technique segment of intensity rapid decrement can relatively easily be reached, also, the size expansion device namely the ultrasonic wave damper
Its installation and removal is all very convenient, it is important that it can also be reused.
The technical scheme of this case has dissolved its application related one to dimethyl silicone polymer substrate addressed above totally
Row technical barrier.Based on this case scheme, this kind very cheap polydimethyl siloxane material is just possible in the micro-fluidic chip
Prepare, production, using etc. field play bigger effect.
Its highly integrated structure of this case micro-fluidic chip, is measured the need for determining it among test application to Serum samples
Smaller, this advantageously reduces the body and mind damage of 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；Figure
Example in arrow indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, the flowing of its test liquid stream
Direction.It is depicted without being also depicted without the elastic clip in the ultrasonic wave damper, legend in legend.
Embodiment
In this case that Fig. 1 is shown embodiment, the example is characterized in, the structure of the device includes multichannel micro-fluidic core
Piece, the structure of the micro-fluidic chip includes the substrate 1 and cover plate 2 of installing bonded to each other 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 of simple drilling technology formation, substrate 1 bonded to each other being installed together and the cover plate 2 are built into jointly
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 1 with being somebody's turn to do
The interface zone bonded to each other of cover plate 2, its side of the window is blocked by the cover plate 2 and opposite side is opened, the structure position of the window
Put be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are the sample introduction positioned at different structure position respectively
Liquid pool and terminal liquid pool are held, there is the one or more sample introduction end liquid pool position of sample introduction end 5 of the micro-fluidic chip,
The sample introduction end 5 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, at the end of the micro-fluidic chip
6 positions are held then to have a terminal liquid pool, the terminal 6 is referred to when the actual sample introduction of the micro-fluidic chip is tested in its chip
The terminal location of liquid flowing, the terminal 6 is located remotely from each other with the sample introduction end 5, one end of the pipeline and the institute positioned at sample introduction end 5
Sample introduction end liquid pool UNICOM is stated, the other end of the pipeline and the terminal liquid pool of the terminal 6 positioned at the micro-fluidic chip join
It is logical, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and reference
Electrode, the order refers to its locations of structures of the reference electrode closer to the position of terminal 6, and the backward refers to institute
Reference electrode locations of structures is stated closer to the position of sample introduction end 5, the working electrode is by conductive electrode and is attached to this and leads
The gold size sensitive membrane for having embedded tumor markers antibody on conductive electrodes is constituted, and parallel construction is presented in the construction of the pipeline, described
It is made up of in the pipeline of parallel construction eight lateral parallel connections, its appearance profile of the pipeline of the presentation parallel construction is approximate
In the profile of parallel circuit, the quantity of the working electrode is eight, and the installation position of eight working electrodes is respectively positioned at described
In eight laterals, and, the tumor markers antibody in the sensitive membrane structure of its top layer gold size of eight working electrodes is respectively
The eight kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen, eight kinds of antibody materials are tumour mark respectively
Will thing antibody CA199, CA153, CA125, CA242, AFP, CEA, EB and β-HCG, the antigen is the antigen of broad sense,
The antibody is the antibody of broad sense, and its material of the working electrode is argent material, gold material, carbon material or thermal decomposition
Column, sheet or thread is presented in conducting polymer material, the working electrode its pattern, and its material of the substrate 1 is poly dimethyl
Siloxanes material, its surface of substrate 1 is the surface of primary form, the surface of the primary form its be intended to refer to not pass through
The surface of the primary form of any surface chemical modification or the material of any surface chemical modification, the structure of the device is also including micro-
Type ultrasonic transducer 4, and, higher-order of oscillation electric signal transmission cable 3, the one of the higher-order of oscillation electric signal transmission cable 3
End links together with the miniature ultrasonic transducer units 4, and the miniature ultrasonic transducer units 4 are installed in the micro-fluidic chip with attaching
Cover plate 2 or substrate 1 the neighbouring terminal 6 position；Its major function of the miniature ultrasonic transducer units 4 is in miniflow
When controlling the actual sample introduction of chip and testing, the ultrasonic wave launched using it reduces the boundary between sample solution and the inwall of the pipeline
Face tension force, 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 its
Interfacial tension between difference between interfacial tension and the terminal 6 its interfacial tension, the micro-fluidic chip two ends 5,6
Difference can form pressure gap between the two ends 5,6 of the micro-fluidic chip, and the pressure gap can drive sample solution to institute
Terminal 6 is stated to flow；Its function of the miniature ultrasonic transducer units 4 also includes checking with its ultrasonic wave launched contained in sample
Large biological molecule its absorption on the inner surface of pipeline, and then check the substrate 1 of the dimethyl silicone polymer material its body
With respect to the effect of swallowing up of the large biological molecule；Its function of the substrate 1 of the dimethyl silicone polymer material includes and cover plate 2 and work
Make electrode and the micro-fluidic chip is together built to electrode and reference electrode, the dimethyl silicone polymer material that is soft and having elasticity
Substrate 1 its function also include, with its property to the strong absorption of ultrasonic wave, ultrasonic wave being absorbed strongly, and thereby at this
The micro-fluidic chip terminal 6 is to the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end 5；With
And, ultrasonic wave damper, its profile of ultrasonic wave damper is in block, tabular or bar-shaped；Its material of the ultrasonic wave damper is
Microcellular rubber, sponge rubber or microporous polyurethane elastomer；And, elastic clip, the elastic force accompanies two clamping limbs, each folder
The medial surface of gripping arm is relative with the medial surface of another clamping limb, and the medial surface of one of clamping limb is close to the micro-fluidic chip
Sample introduction end 5, the medial surface of another clamping limb is close to the ultrasonic wave damper, and the elastic clip is micro- by this with its gripping strength
The sample introduction end 5 of fluidic chip and the ultrasonic wave damper are clipped together bonded to each otherly, and the ultrasonic wave damper is with its material institute band
The strong consumption to ultrasonic wave come, is further supported in micro-fluidic chip terminal 6 to having between the sample introduction end 5
The rapid decrement of ultrasonic intensity is realized within the short distance of limit, and thereby further expand described its interfacial tension of sample introduction end 5 with
Difference between described its interfacial tension of terminal 6.
It is depicted without being also depicted without the elastic clip in the ultrasonic wave damper, legend in legend.
Arrow in legend indicate the micro-fluidic chip its in actual motion, by pressure at two ends difference drive, its test liquid
The flow direction of 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；It can also be customized to ultrasonic transducer producer or to cable
Specialized factory customizes.
Involved higher-order of oscillation electric signal generator market has the product close to needs commercially available；It can also be customized to relevant speciality producer.
The multi-channel electrochemical work station can be with commercially available；The multi-channel electrochemical work station can also be according to specific
Whereabouts specialised manufacturers are needed to customize.
Each working electrode in this example structure and electrode and reference electrode via each special cable or can be said respectively
Get 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 hydrophobic
Capillary form pipeline.
In view of this case pipeline of the presentation parallel construction its form that related text expresses that it is described above is clear enough
It is 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；It is immunized described in this case multiple
Compound refers to the immune complex of broad sense.
Claims (10)
1. the classifiable tumor markers in detecting micro flow control chip device that women is applicable, it is characterised in that the structure of the device
Including multichannel micro-fluidic chip, the structure of the micro-fluidic chip includes the substrate and cover plate of installing bonded to each other together, described
Substrate 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 formation channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etch
Technique or the window structure of simple drilling technology formation, substrate bonded to each other being installed together are built into jointly with the cover plate
Micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline is located at the substrate and the lid
Piece interface zone bonded to each other, its side of the window is blocked by the cover plate and opposite side is opened, and the locations of structures of the window is exactly
The locations of structures of the liquid pool, the liquid pool has two kinds, and two kinds of liquid pools are the sample introduction end liquid pool positioned at different structure position respectively
And terminal liquid pool, the sample introduction end position of the micro-fluidic chip has one or more sample introduction end liquid pool, the sample introduction
End refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, then has in the terminal location of the micro-fluidic chip
One terminal liquid pool, the terminal refers to the terminal of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested
Position, the terminal is located remotely from each other with the sample introduction end, one end of the pipeline and the sample introduction end liquid pool UNICOM positioned at sample introduction end, should
The other end of pipeline and the terminal liquid pool UNICOM of the terminal positioned at the micro-fluidic chip, and, sequentially or backward
The working electrode that is respectively installed in the pipeline on diverse location and to electrode and reference electrode, the order refers to described
Its locations of structures of reference electrode is closer to the terminal location, and the backward refers to the reference electrode locations of structures closer to institute
State sample introduction end position, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumor markers
The gold size sensitive membrane of antibody is constituted, and parallel construction is presented in the construction of the pipeline, and the pipeline in parallel construction is by eight branched pipes
Road parallel connection is constituted, and its appearance profile of the pipeline of the presentation parallel construction is similar to the profile of parallel circuit, the work electricity
The quantity of pole is eight, and the installation position of eight working electrodes is located in eight laterals respectively, and, this eight
Tumor markers antibody in the sensitive membrane structure of its top layer gold size of working electrode is respectively can specificity knot to tumor markers antigen
Close eight kinds of tumor markers antibody materials, eight kinds of antibody materials be respectively tumor markers antibody CA199, CA153, CA125,
CA242, AFP, CEA, EB and β-HCG, the antigen are the antigen of broad sense, and the antibody is the antibody of broad sense, the work
It is argent material, gold material, carbon material or thermal decomposition conducting polymer material, the working electrode to make electrode its material
Column, sheet or thread is presented in its pattern, and its material of the substrate is dimethyl silicone polymer material, and its surface of the substrate is primary
The surface of form, the surface of the primary form its be intended to refer to by any surface chemical modification or any surface chemistry not change
Property the material primary form surface, the structure of the device also includes miniature ultrasonic transducer units, and, higher-order of oscillation electricity
Signal transmission cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable link together, and this is micro-
Type ultrasonic transducer is installed in the position of the cover plate of the micro-fluidic chip or the neighbouring terminal of substrate with attaching；This is miniature super
Its major function of acoustic wave transducer is that in the test of micro-fluidic chip actual sample introduction, the ultrasonic wave launched using it reduces sample
Interfacial tension between the inwall of solution and the pipeline, can be compatible, also, utilizes the sample introduction end and the end
Difference in the distance between end and the miniature ultrasonic transducer units installation position difference and its ultrasonic intensity experienced,
Difference between its interfacial tension of sample introduction end described in induced synthesis and the terminal its interfacial tension, the micro-fluidic chip two ends it
Between interfacial tension difference can form pressure gap between the two ends of the micro-fluidic chip, the pressure gap can drive sample molten
Liquid is to the end flow；Its function of the miniature ultrasonic transducer units is also contained in sample including being checked with its ultrasonic wave launched
Its absorption on the inner surface of pipeline of some large biological molecules, 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；Its function of the substrate of the dimethyl silicone polymer material includes and cover plate and work electricity
Pole and the micro-fluidic chip is together built to electrode and reference electrode, the base of the dimethyl silicone polymer material that is soft and having elasticity
Piece its function also includes, with its property to the strong absorption of ultrasonic wave, ultrasonic wave being absorbed strongly, and thereby micro-fluidic at this
The chip terminal is to the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end；And, ultrasonic wave
Damper, its profile of ultrasonic wave damper is in block, tabular or bar-shaped；Its material of the ultrasonic wave damper be microcellular rubber,
Sponge rubber or microporous polyurethane elastomer；And, elastic clip, the elastic force accompanies two clamping limbs, the inner side of each clamping limb
Face is relative with the medial surface of another clamping limb, and the medial surface of one of clamping limb is close to the sample introduction end of the micro-fluidic chip,
The medial surface of another clamping limb is close to the ultrasonic wave damper, the elastic clip entering the micro-fluidic chip with its gripping strength
Sample end and the ultrasonic wave damper are clipped together bonded to each otherly, the ultrasonic wave damper with its material brought to ultrasonic wave
Strong consumption, further supports to realize within micro-fluidic chip terminal to the limited short distance between the sample introduction end
The rapid decrement of ultrasonic intensity, and thereby further expand described its interfacial tension of sample introduction end and the terminal its interfacial tension it
Between difference.
2. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, it is capillary channel that the pipeline, which includes the lateral,.
3. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, the thermal decomposition conducting polymer be by polyimides or polyacrylonitrile be heat-treated through anoxybiotic after the electric conductivity material that is formed
Material.
4. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, 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. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.
6. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, its material of the cover plate in structure is dimethyl silicone polymer material.
7. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, the distance between the terminal and the sample introduction end between 3 centimetres and 10 centimetres, the cover plate and substrate its
Thickness is between 1.0 millimeters and 5.0 millimeters.
8. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, the microcellular rubber is micropore silicon rubber or micropore fluorubber.
9. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, 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. the classifiable tumor markers in detecting micro flow control chip device that women according to claim 1 is applicable, it is special
Levy and be, its specified ultrasonic wave transmission power of the miniature ultrasonic transducer units is between 2 milliwatts and 2000 milliwatts, and this is miniature
The frequency of its ultrasonic wave operationally launched of ultrasonic transducer is between 100KHz and 12MHz.
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Cited By (1)
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