CN106680501A - Convenient disassembling universal multi-tumor marker joint inspection chip device - Google Patents
Convenient disassembling universal multi-tumor marker joint inspection chip device Download PDFInfo
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- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
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Abstract
The invention relates to a convenient disassembling universal multi-tumor marker joint inspection chip device, and belongs to the field of analysis and tests. The polydimethylsiloxane (PDMS) is used to manufacture a substrate for the universal microfluidic chip used for joint inspection of more than ten tumor markers, the device has obvious advantages, but a plurality of difficult problems needing to be solved also exist, and the scheme aims at solving the problems. The key point of the scheme is as follows: the PDMS with an original ecological surface is chosen for the substrate, a hinge type clamp equipped with a minitype ultrasonic transducer is manually fastened and located on a terminal or the adjacent position of a sample liquid flow of the microfluidic chip to reduce the interfacial tension through the ultrasonic wave, the compatibility of an interface between a solid phase and a liquid phase is greatly improved, meanwhile, the strong power of absorbing the ultrasonic wave of the PDMS is utilized to rapidly decrease the ultrasonic intensity progressively within a short distance, so that the difference of interfacial tensions is formed at the two ends of the chip, and the sample liquid flow flows towards the direction of the terminal along with an originally hydrophobic capillary channel.
Description
Technical field
The present invention relates to a kind of general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled, belong to analysis test neck
Domain.
Background technology
Tumor markerses (tumor marker, TM) are referred in the generation and breeding of tumor, produced by tumor cell itself
It is raw or by body to tumor cell react and produce, reflect tumor exist and grow a class material, including protein,
Hormone, enzyme (isozyme) and oncoprotein etc..Tumor markerses in chemical examination blood samples of patients or body fluid, can be in cancer screening
Early discovery tumor, and observe the curative effect of oncotherapy and judge patient's prognosis.At present clinically conventional tumor markerses have:
(1) alpha-fetoprotein (AFP) is the mark of the tumors such as primary hepatocarcinoma, carcinoma of testis, ovarian cancer;(2) carcinoembryonic antigen (CEA) is digestion
The mark of the tumors such as system tumor, pulmonary carcinoma, breast carcinoma;(3) CA125 (CA125) is the mark of the tumors such as ovarian cancer;
(4) CA153 (CA153) is the mark of the tumors such as breast carcinoma;(5) CA19-9 (CA19-9) is digestive system tumor
Mark;(6) CA724 (CA724) is the mark of the tumors such as gastric cancer, ovarian cancer;(7) carbohydrate antigen 242 (CA242)
For the mark of digestive system tumor;(8) CA50 (CA50) is the mark of the tumors such as digestive system tumor, breast carcinoma, pulmonary carcinoma
Thing;(9) CYFRA21-1 (Cy211) is the mark of the tumors such as nonsmall-cell lung cancer;(10) neuronspecific enolase (NSE)
For the mark of the tumors such as small cell lung cancer, neuroendocrine tumor;(11) prostate specific antigen (PSA) is carcinoma of prostate
Tumor markerses;(12) human chorionic gonadotropin (HCG) is that ECC, trophoblastic tumor (choriocarcinoma, hydatidiform mole) etc. are swollen
The mark of tumor;(13) Elityran (TG) is the mark of thyroid carcinoma;(14) ferritin (SF) is digestive system tumor, liver
The mark of the tumors such as cancer, breast carcinoma, pulmonary carcinoma;(15) B2M (β 2-MG) is in chronic lymphocytic leukemia, lymph
Raise in the patient body fluids such as tumor, myeloma, pulmonary carcinoma, thyroid carcinoma, nasopharyngeal carcinoma;(16) squamous cell antigen (SCC) be cervical cancer,
The tumor markerses such as lung squamous cancer, the esophageal carcinoma.The tumor markerses overwhelming majority for clinically detecting at present is not only present in malignant tumor
In, exist in benign tumor, embryonal tissue, even in normal structure.Therefore, tumor markerses have dynamic chek and multinomial
Joint inspection is more valuable.So for numerous tumor markerses, clinically how to selectDifferent tumors understands some phases
To special tumor markerses, such as CA153 often occurs in breast carcinoma;CEA often occurs in intestinal cancer, gastric cancer;CA19-9 is often occurred in
Intestinal cancer, cancer of pancreas;CA125 often occurs in ovarian cancer etc..Clinician can be according to the different mark of different tumor examinations.
Same tumor or different types of tumor can have one or more tumor markerses exceptions;Same tumor markerses can be in difference
Tumor in occur.To improve the additive diagnostic value of tumor markerses and determining which kind of mark can be supervised as the follow-up after treatment
Index is surveyed, tumor markerses joint-detection, the complementary tumor markerses 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 markerses 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 is may refer to soon
Before the monograph " diagram Microfluid based Lab on a chip " that goes out, the monograph publishes via Science Press, and the monograph is for micro-fluidic
Past of technology, now, and, vision of the future etc. aspect, suffer from detail, be deep into long discussion of detail.
So, the focal issue that this case that to have a talk below pays special attention to.
The basic framework of micro-fluidic chip, including the substrate for being etched with small fluid course and the cover plate for fitting together therewith,
Small fluid course on the substrate, before assembling upper cover plate, it is apparent on see to be exactly some micro-channel, be when 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 together described small fluid course;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 determines 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 the micro-channel on piece is key factor;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, piezoid, sheet glass, high polymer such as polydimethylsiloxane, polymethyl methacrylate, Merlon etc.
Deng;Certainly, the selection of substrate can be with identical with the selection of cover plate, it is also possible to differs;From material consumption, manufacture difficulty and
Application popularization prospect etc. aspect from the point of view of, there is not little difference between these materials, especially the selection of that substrate, affect compared with
Greatly.
In various substrate making materials, polydimethylsiloxane, i.e. PDMS, comparatively very easy molding, 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 suppressed or etched thereon, and the cover plate made with glass or the cheap material such as polypropylene or other plastic sheets is engaged,
It is seemingly a kind of more satisfactory selection;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, on this material micro-channel is built, if not carrying out the modified operation for the micro-channel surface, then,
Integral installation is covered after cover plate with after completing, because of structure in the micro-channel its inner surface occupy most liquid circulation
The inner surface in road, then, the PDMS micro-channel inner surface its strong hydrophobic property is deciding factor, and it can cause class
Be similar to 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 surfaciesThat falls nor this problem, structure
It is another problem into serious technical puzzlement:PDMS polymer molecules tool inside this PDMS material substrate its body phase
There is the characteristic for diffusing to the surface automatically, migrating, this substrate body phase inside PDMS polymer molecules are diffused to the surface, moved automatically
The characteristic of shifting, by cause through that micro-channel of surface modifying and decorating its inner surface it is modified after state can not maintain foot
The enough long time, being held time of the micro-channel after that is surface-modified its inner surface state be substantially only sufficient to, and to complete experiment indoor
The time of portion's test experiments needs;In other words, PDMS micro-channel inner surfacies of or surface modification modified through surface, its
After modified or say the apparent condition formed after modification can not be lasting, but soon automatically tend to or say become surface again and change
Apparent condition before property, returns to the strong hydrophobic apparent condition of that script in the shorter time, then, just think,
Such micro-fluidic chip can make in a large number, mass storage, be widely popularized, and answer is it is obvious that is, impossible.
Micro-channel on this PDMS material, does not do if surface modification, cannot pump similar to the fine liquid stream of polar solvent of aqueous solution
Send and pass through, chip also just cannot be used;And if having done surface modification, the state after its modification cannot be persistently kept again, also
Being equally cannot 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 cannot persistently, chip cannot in a large number make, deposit and then be widely popularized and such a make this area numerous specially in a large number
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 days 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 biomacromolecule, and
And, the depression that these adsorbed biomacromolecules can also be further further on PDMS surfaces gradually falls into gradually deep, directly
It is trapped within the body phase of PDMS substrates to heavy, in fact, this process is partly also due to inside PDMS material body phase
Polymer molecule have diffuse to the surface, travel motion is caused;Such case, it is also possible to explain from another angle, i.e.
Continuously from inside PDMS body phases to its diffusion into the surface, migration those polymer molecules, its motion result, be by
Gradually those are involved within the body phase of PDMS substrates by the biomacromolecule of surface adsorption, briefly, these are inhaled
Attached biomacromolecule is exactly to be swallowed up by PDMS substrate body phases;So, this PDMS substrates body phase swallows up biomacromolecule
Phenomenon, the impact caused by it necessarily causes to be related to the severe deviations of all kinds of test data of experiment of biomacromolecule.
As described above, the problem of PDMS substrates is, its not only surface adsorption biomacromolecule, and swallow up biomacromolecule,
So, as the biomacromolecule of experiment test object, its disappearance will not stop because surface saturation is adsorbed, but, no
It is disconnected adsorbed, also constantly swallowed up.
With regard to PDMS substrates in related experiment test process its body phase constantly swallow up test associated biomolecule macromole phenomenon, separately
It is to say that one kind is explained, there are substantial amounts of Minute pores in PDMS body phases, associated biomolecule macromole by after surface adsorption, depression
Into these Minute pores, and then swallowed up;However, inventor thinks, those can allow the air point of miniature scale
Son clamp-ons the Minute pores therebetween, and being not equal to them also can directly allow that the biomacromolecule of relative large scale is entered, and two
Person's difference on yardstick is huge, must not make sweeping generalizations.Explanation is bypassed, in any case, as dependence test the biology of object is analyzed
Macromole is adsorbed by PDMS substrate micro-channel inner surface, and then is constantly swallowed up by PDMS substrate body phases, and this is known objective
The phenomenon of presence.
In order to prevent this PDMS substrate bodies relative to the effect of swallowing up of biomacromolecule, can be from containment PDMS surfaces opposite
The absorption of thing macromole addressing, method be aiming at the PDMS material surface be chemically modified it is modified, for
For PDMS is for the situation of substrate material, exactly modified, Jing Guohua is chemically modified to the surface of described micro-channel part
The micro-channel inner surface of modification is learned, its absorption to biomacromolecule can be contained, and then avoid biomacromolecule quilt
PDMS substrate body phases are swallowed up;But, or that old problem, that is, the chemical modification on PDMS material surface changes
Apparent condition after property cannot persistently keep, the polymer molecule inside the PDMS substrate body phases its diffuse to the surface automatically,
The process of migration, soon can become that again script strong and dredge through the micro-channel inner surface state that surface chemical modification is modified
The state of water and strong adsorption biomacromolecule, in other words, no matter how professionals in the field turn from side to side, the PDMS
Substrate its micro-channel inner surface 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 the absorption process of the PDMS substrate micro-channel inner surfacies to biomacromolecule, and then prevents PDMS substrate body phases
The effect of swallowing up to biomacromolecule so that related chip manufactured goods are able to maintain that a prolonged enough, rational shelf-life,
It is exactly a very thorny difficult problem.The difficult problem makes the numerous specialties in this area as another difficult problem addressed above, equally
Personnel are entangled with for a long time, perplex, and the difficult problem is equally the highly difficult problem that its obvious technology barrier can not despise.The hardly possible
Also be present many days in topic, so far, also not yet properly settled.
The content of the invention
The technical problem to be solved is to provide a package solution, solves what is addressed above totally
Two difficult problems, also, the solution is applied to build a kind of new crowd's health check-up is blanket can be compared for ten kinds
More typical primary tumor mark carries out simultaneously examination, while the micro flow control chip device of detection.
The present invention solves the technical problem by following scheme, and the device that the program is provided is a kind of general type conveniently disassembled
Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus, program main points are that the structure of the device includes multichannel micro-fluidic chip, should
The structure of micro-fluidic chip includes the substrate installed together bonded to each other and cover plate, and the substrate and cover plate are plate object or piece
Contain the channel structure formed via mould pressing process or etching technics, the substrate in shape thing, that face towards the cover plate of the substrate
Also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics or simple drilling technology formation
Window structure, substrate being installed together bonded to each other has been built into containing pipeline configuration and has been attached thereto jointly with the cover plate
Liquid pool structure micro-fluidic chip, the locations of structures of the pipeline is located at the substrate and cover plate interface zone bonded to each other, should
Its side of 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, described
Liquid pool has two kinds, two kinds of liquid pools respectively be located at different structure position sample introduction end liquid pool and terminal liquid pool, the micro-fluidic core
The sample introduction end position of piece has one or more sample introduction end liquid pool, and the sample introduction end refers to the micro-fluidic chip reality
The injection end position of detected solution during sample introduction, then has a terminal liquid pool, the end in the terminal location of the micro-fluidic chip
End 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 phase
Mutually away from one end of the pipeline is with the sample introduction end liquid pool UNICOM for being located at sample introduction end, the other end of the pipeline and positioned at the miniflow
The terminal liquid pool UNICOM of the terminal of control chip, and, sequentially or backward ground is respectively installed in the pipeline difference positions
The working electrode put and to electrode and reference electrode, the order refer to its locations of structures of the reference electrode closer to
The terminal location, the backward refers to the reference electrode locations of structures closer to the sample introduction end position, the work electricity
Pole is made up of conductive electrode and the gold size sensitive membrane for having embedded tumor markerses antibody being attached on the conductive electrode, should
The construction of pipeline is presented parallel construction, and the pipeline in parallel construction is made up of 15 lateral parallel connections, and the presentation is simultaneously
The pipeline its appearance profile of connection construction is similar to the profile of parallel circuit, and the quantity of the working electrode is 15, and this ten
The installation position of five working electrodes is located in 15 laterals respectively, and, 15 its top layers of working electrode
Tumor markerses antibody in gold size sensitivity membrane structure is respectively the 15 kinds of tumors that can be specifically bound to tumor markerses antigen
Mark antibody materials, 15 kinds of antibody materials be respectively tumor markerses antibody A FP, CEA, CA242, CA125, CA199,
CA153, CA724, CA50, NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, it is described anti-
Original is the antigen of broad sense, and the antibody is the antibody of broad sense, the working electrode its material be argent material, gold material,
Carbon material or thermal decomposition conducting polymer material, the working electrode its pattern is presented column, lamellar or thread, the substrate its
Material is polydimethylsiloxane material, and its surface of the substrate is the surface of primary form, the surface of the primary form its be meant to
Be not through any surface chemical modification or the surface of the primary form of the material of any surface chemical modification, the device
Structure also includes hinge-type fixture, the hinge-type fixture its appearance profile likeness in form hinge, and the hinge-type fixture is by being hinged one
Two hinges and a fastening screw through two hinges and one for rising match with the fastening screw and with the fastening
The nut for fastening and hand break loose manually that screw connects together is constituted, and two hinges of the hinge-type fixture are respectively with its end
The micro-fluidic chip is mutually drawn close and clamped to the tip, and the tip is positioned at the locations of structures of the neighbouring described terminal of the micro-fluidic chip
On, fixation is attached in a hinge at least in and is equiped with miniature ultrasonic transducer units, and, higher-order of oscillation telecommunication
Number transmission cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable link together;The hinge
Formula fixture provides a function of facilitating the device to disassemble;The miniature ultrasonic transducer units its major functions is in micro-fluidic chip
When actual sample introduction is tested, the ultrasound wave launched using it is reducing the interface between sample solution and the inwall of the pipeline
Power so as to can be compatible, also, using the sample introduction end and the terminal and the miniature ultrasonic transducer units installation position it
Between distance difference and the difference on its ultrasonic intensity for being experienced, its interfacial tension of sample introduction end and institute described in induced synthesis
The difference between terminal its interfacial tension is stated, the interfacial tension difference between the micro-fluidic chip two ends can be in the micro-fluidic chip
The two ends between form pressure gap, the pressure gap can drive sample solution to the end flow;The miniature ultrasonic is changed
Can device its function also include that the ultrasound wave launched with it checks biomacromolecule contained in sample its table in the pipeline
Absorption on face, and then check swallow up effect of its body phase of the substrate of the polydimethylsiloxane material to the biomacromolecule;Institute
Stating the substrate of polydimethylsiloxane material its function includes with cover plate and working electrode and to electrode and reference electrode together building
The micro-fluidic chip, it is soft and have the substrate of the polydimethylsiloxane material of elasticity its function also include it is strong to ultrasound wave with it
The property of strong absorption, is absorbed strongly to ultrasound wave, and thereby in the micro-fluidic chip terminal to having between the sample introduction end
The rapid decrement of ultrasonic intensity is realized within the short distance of limit.
The tumor markerses antibody is the antibody of broad sense, and the antibody of the broad sense is referred to possess antibody function or is functionally similar to
Immune complex or immune compound can be combined and be formed in antibody with the involved tumor markerses of various corresponding clinics
The material of the analog of thing;The tumor markerses antigen is the antigen of broad sense, and the antigen of the broad sense is referred to can be using corresponding
Antibody or be functionally similar to antibody material carry out enzyme mark detection it is various clinic involved by the tumor-marker for needing to differentiate, detect
Thing.
Described miniature ultrasonic transducer units can certainly be all installed in two hinges;But, only installing one is micro-
Type ultrasonic transducer is dealt with enough and is used.
The word of hinge one art-recognized meanings of itself are known.
The gold size sensitive membrane is to be sufficiently mixed shitosan gold size solution and tumor markerses antibody-solutions uniformly, uses point sample instrument
Point sample coats specified structure position, and forms its drying and forming-film.Tumor markerses antibody in the gold size sensitive membrane is equal
For horseradish peroxidase or the tumor markerses antibody of glucose oxidase labelling, the gold size sensitive membrane is included as fixing
Above-mentioned each tumor markerses antibody and introduce complementary medium therein, the complementary medium for example shitosan, 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, but, for prepare liquid sample and the consideration of the aspects such as reagent loss is reduced less as far as possible, the pipeline is including described
The passage of the preferred capillary level of lateral, the passage of the capillary level implies that the interior of the capillary tube on its internal diameter and ordinary meaning
The suitable passage in footpath.The shape of cross section of its inner passage of capillary tube 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
With the extension of pipeline, the shape of cross section of different parts can also allow to be different shapes the interior shape of tubule.Only with regard to hair
For the word of tubule one, its art-recognized meanings is known.
What is be related in structure is minute sized electrode to electrode and reference electrode, and its electrode shape may each be arbitrarily choosing
Fixed shape, the arbitrarily selected shape such as column, lamellar, strip or thread etc..It is described to electrode and the ginseng
It is known than the art-recognized meanings of the electrode vocabulary of itself.
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, the inner chamber of each liquid pool its shape 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 for the word of ultrasonic transducer one art-recognized meanings of itself of professional only with regard to to(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 little to only with millimeter calculate magnitude.
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 ground, 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 device can also include higher-order of oscillation electric signal generator;Its is another for the higher-order of oscillation electric signal transmission cable
One end can be connected with the higher-order of oscillation electric signal generator.
The involved higher-order of oscillation electric signal generator technology of itself, 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 the miniature ultrasonic transducer units its specified ultrasonic emitting power be between 5 milliwatts and 9000 milliwatts it
Between;The preferred scope of the frequency of the miniature ultrasonic transducer units its ultrasound wave operationally launched be between 100KHz with
Between 12MHz.
This case device can further include some adnexaes, the adnexa 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
To electrode and reference electrode etc., can respectively via corresponding special the corresponding interface got lines crossed with the multiple tracks electrochemical workstation
Coupled.It is described that special to get lines crossed be for each electrode is carried out into phase with each the corresponding interface of the multiple tracks electrochemical workstation
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, and according to actual needs, the micro-valve can be installed in any need position to be mounted in the microfluidic chip structure;
For the professional of micro fluidic chip technical field, the art-recognized meanings of itself are known to the word of micro-valve one;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 for using, 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 for using, it is however recommended to or to say the preferred length its scope 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 other appropriate process coatings is quick
Sense film, its thicknesses of layers can allow the thickness for being the sample measuring liquid the treated generation electrical signals response of any setting, but, push away
In other words conj.or perhaps preferred thickness is between 10 nanometers and 200 nanometers to 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, polydimethylsiloxane, etc., in order to make smaller size of micro-fluidic chip, such as do
Into the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and realize to ultrasound wave in the extremely short distance
Extremely fast decay, it may be preferred to polydimethylsiloxane is used as cover plate.Certainly, select on large-sized micro-fluidic chip
It is used as the cover plate using polydimethylsiloxane, 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.
The cover plate and substrate its thickness can allow the thickness for being easy to assemble for being any setting, the thickness of recommendation or say preferably
Thickness be between 1.0 millimeters and 5.0 millimeters.Less thickness is conducive to material-saving.
The using method of this case micro-fluidic chip:
Proposed first based on this case and the first public New stream driving principle, this case micro-fluidic chip its application running
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 the micro-fluidic chip two ends caused by the ultrasound wave, drives liquid
Stream flows in the capillary channel of the ten Five-channels micro-fluidic chip, using multi-channel electrochemical analyzer device respectively to 15 kinds
Tumor markerses carry out joint-detection.
The concrete detection of this case micro-fluidic chip is as follows using step:
1st, blood serum sample liquid is added in micro-pipe road, in the case where the ultrasound wave drives, various tumor markerses antigen molecules are each
In passage on electrode surface gold size sensitive membrane embedding corresponding horseradish peroxidase-labeled tumor markerses specific antibody
Capture.
2nd, the tumor markerses antigen in the tumor markerses specific antibody and blood serum sample of horseradish peroxidase-labeled is formed
Immune complex.
3rd, multi-channel electrochemical analyzer is adopted, the electron mediators such as catechol is added, using the above-mentioned reaction of amperometric detection
The curent change for causing, is derived from the species and content of various analytes.
4th, result is carried out into comprehensive analysis, comprehensive diagnos is carried out to tumor markerses antigen.
It is an advantage of the invention that its close position positions the hinge-type fixture in the terminal of the micro-fluidic chip, with
The miniature ultrasonic transducer units installed, the low-power launched using it, high-frequency band are attached in the hinge-type fixture its hinge
Ultrasound wave so that without surface chemical modification or surface chemical modification the strong hydrophobic micro-fluidic chip internal pipeline its
The compatibility between tube wall and test object aqueous solution is significantly increased, and this is test liquid stream by possible there is provided a reality
Property;Meanwhile, using its strong absorbability to ultrasound wave of polydimethylsiloxane substrate, in shorter distance, also
Be, from the terminal to the very short distance of the only several centimeters yardstick the sample introduction end in, reach the quick of ultrasonic intensity
Successively decrease, the difference of the interfacial tension is thereby caused at the two ends of the micro-fluidic chip, and then, using between the two ends
Pressure gap between its two ends for being formed of the difference of interfacial tension, drives test liquid stream originally strong hydrophobic in such a
Capillary channel in the terminal direction flowing.By this case liquid stream drive scheme, entirely without must be to the polydimethylsiloxanes
The substrate and its internal pipeline of alkane material carries out any surface chemical modification or chemical modification, has altogether dispensed with the surface chemistry and has repaiied
The laborious procedures of decorations or chemical modification;And altogether dispense with the equipment of traditional Micropump etc;On the other hand, this is low
The ultrasound wave of power, high-frequency band, additionally it is possible to contain the biomacromolecule in sample in the literalness naked polydimethylsiloxanes
Absorption on alkyl piece its inner surface of pipeline, and then contain the polydimethylsiloxane substrate its body phase to the biomacromolecule
The effect of swallowing up;The Reversible binding thing of the antigen, antibody and antigen and antibody is all belonging to described biomacromolecule certainly
Type;Because described adsorption and the described effect of swallowing up effectively are contained, therefore, dependence test result will more
Practical situation can objectively be reflected;The effect of the low-power, high-frequency band ultrasound wave, also includes facilitating antigen, antibody certainly
Between Reversible binding reaction quickly reach, this cause dependence test operation can be to complete than speed faster.
Due to the surface chemical modification for the polydimethylsiloxane 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 presence, then, the polydimethylsiloxane
Substrate its body phase interior polymer molecule constantly diffuses to the surface automatically, migrate caused by its institute to the surface chemical modification layer or
The damaging influence of chemically modified layer is not also just present.
The technical scheme of this case has dissolved its application related one to polydimethylsiloxane substrate addressed above totally
Row technical barrier.Based on this case scheme, the very cheap polydimethyl siloxane material of this kind is just possible in the micro-fluidic chip
Preparation, production, application etc. field plays bigger effect.
The miniature ultrasonic transducer units have been installed in fixation in the hinge-type fixture its hinge in this case structure, and the structure is provided
One function of facilitating the device to disassemble, thus, the hinge-type fixture is together with miniature ultrasonic transducer units appended in its hinge
Just easily can be mutually disengaged with the micro-fluidic chip, then, the component that the part can freely depart from just can circulate benignly ground
Reuse many times;The architectural feature is conducive to saving the use cost of the device.
The framework of this case micro-fluidic chip its large-scale integrated, determines it among actual test application, to Serum samples
Requirement it is less, this body and mind for contributing to reducing related subject is damaged.
Description of the drawings
Fig. 1 is its rough outside side view of this case device.
In figure, 1 is hinge-type fixture, and 2 indicate linkwork positions, and 3 is fastening screw, and 4 is higher-order of oscillation telecommunication
Number transmission cable, 5 is miniature ultrasonic transducer units, and 6,11 is respectively two different hinges of locations of structures, and 7 is cover plate, 8
It is substrate, 9 is sample introduction end, and 10 is terminal, and 12 is nut;The hinge-type clamp structure in legend is only the legend illustrated
Structure, actual its structure of the hinge-type fixture is not limited to the legend hinge-type clamp structure;It is micro- that arrow in legend indicates this
Fluidic chip its in actual motion, by pressure at two ends difference drive, the flow direction of its test liquid stream.
Specific embodiment
In this case that Fig. 1 is shown embodiment, the example its main points are that the structure of the device includes multichannel micro-fluidic core
Piece, the structure of the micro-fluidic chip includes the substrate 8 installed together bonded to each other and cover plate 7, the substrate 8 and cover plate 7
Plate object or tablet are, that face towards the cover plate 7 of the substrate 8 is contained and formed via mould pressing process or etching technics
Channel structure, the substrate 8 also containing be connected with the channel structure and pierce the substrate 8 via mould pressing process, etch work
The window structure that skill or simple drilling technology are formed, substrate 8 being installed together bonded to each other is built into jointly with the cover plate 7
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 8 with
The cover plate 7 interface zone bonded to each other, its side of the window is blocked by the cover plate 7 and opposite side is opened, 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 respectively to be located at entering for different structure position
There is the one or more sample introduction end liquid sample end liquid pool and terminal liquid pool, the position of sample introduction end 9 of the micro-fluidic chip
Pond, the sample introduction end 9 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, in the micro-fluidic chip
The position of terminal 10 then have a terminal liquid pool, the terminal 10 refers to when the actual sample introduction of the micro-fluidic chip is tested it
The terminal location of liquid flowing in chip, the terminal 10 is located remotely from each other with the sample introduction end 9, one end of the pipeline with positioned at sample introduction
The end of the sample introduction end liquid pool UNICOM at end 9, the other end of the pipeline and the terminal 10 positioned at the micro-fluidic chip
End 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 refers to its locations of structures of the reference electrode closer to the position of the terminal 10, described inverse
Sequence refers to the reference electrode locations of structures closer to the position of sample introduction end 9, the working electrode by conductive electrode and
The gold size sensitive membrane for having embedded tumor markerses antibody being attached on the conductive electrode is constituted, and the construction of the pipeline is presented in parallel
Construction, the pipeline in parallel construction is made up of 15 lateral parallel connections, the pipeline that parallel construction is presented its
Appearance profile is similar to the profile of parallel circuit, and the quantity of the working electrode is 15, the installing of 15 working electrodes
Position is located in 15 laterals respectively, and, in the sensitive membrane structure of its top layer gold size of 15 working electrodes
Tumor markerses antibody is respectively the 15 kinds of tumor markerses antibody materials that can be specifically bound to tumor markerses antigen, and this ten
Five kinds of antibody materials be respectively tumor markerses antibody A FP, CEA, CA242, CA125, CA199, CA153, CA724, CA50,
NSE, CYFRA21-1, FPSA, TPSA, β-hCG, SCCA and β 2-MG, the antigen is the antigen of broad sense, described
Antibody is the antibody of broad sense, and the working electrode its material is that argent material, gold material, carbon material or thermal decomposition are conductive
Macromolecule material, the working electrode its pattern is presented column, lamellar or thread, and the substrate 8 its material is polydimethylsiloxanes
Alkane material, its surface of substrate 8 is the surface of primary form, the surface of the primary form its be intended to refer to not through any
The surface of the primary form of surface chemical modification or the material of any surface chemical modification, the structure of the device also includes hinge-type
Fixture 1, the hinge-type fixture 1 its appearance profile likeness in form hinge, the hinge-type fixture 1 is by two pages being mutually hinged
Piece 6,11 and a fastening screw 3 through two hinges 6,11 and one match with the fastening screw 3 and with
The nut 12 for fastening and hand break loose manually that the fastening screw 3 connects together is constituted, and the two of the hinge-type fixture 1
The micro-fluidic chip is mutually drawn close and clamped to individual hinge 6,11 respectively with its tip, and the tip is positioned at the neighbour of the micro-fluidic chip
In the locations of structures of the nearly terminal 10, a hinge at least in for example attach in hinge 6 fixation be equiped with it is micro-
Type ultrasonic transducer 5, and, higher-order of oscillation electric signal transmission cable 4, the one of the higher-order of oscillation electric signal transmission cable 4
End links together with the miniature ultrasonic transducer units 5;The hinge-type fixture 1 provides a function of facilitating the device to disassemble;
The miniature ultrasonic transducer units 5 its major functions is the ultrasound wave launched using it when the actual sample introduction of micro-fluidic chip is tested
To reduce the interfacial tension between sample solution and the inwall of the pipeline so as to can be compatible, also, using the sample introduction end
The distance between 9 and the terminal 10 and the installation position of miniature ultrasonic transducer units 5 difference and its experienced it is super
Difference between difference on intensity of acoustic wave, its interfacial tension of sample introduction end 9 described in induced synthesis and the terminal 10 its interfacial tension
It is different, the interfacial tension difference between the micro-fluidic chip two ends 9,10 can the two ends 9,10 of the micro-fluidic chip it
Between form pressure gap, the pressure gap can drive sample solution to flow to the terminal 10;The miniature ultrasonic transducer units 5
Its function also include the ultrasound wave launched with it check biomacromolecule contained in sample its on the inner surface of pipeline
Absorption, and then check swallow up effect of its body phase of the substrate 8 of the polydimethylsiloxane material to the biomacromolecule;It is described
The substrate 8 of polydimethylsiloxane material its function includes with cover plate 7 and working electrode and to electrode and reference electrode together building
The micro-fluidic chip, it is soft and have the substrate 8 of the polydimethylsiloxane material of elasticity its function and also include with it to ultrasound wave
The strong property for absorbing, is absorbed strongly to ultrasound wave, and thereby in the micro-fluidic chip, the terminal 10 arrives the sample introduction end 9
Between limited short distance within realize the rapid decrement of ultrasonic intensity.
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.
The associate members such as the higher-order of oscillation electric signal generator and multiple tracks electrochemical workstation are depicted without in Fig. 1.
Involved hinge-type fixture 1 can carry out simply cutting, the transformation of the way using commercially available hinge;The miniature ultrasonic transducing
Device 5 is commercially available;Commercially available miniature ultrasonic transducer units are various in style, can select as needed to use;The miniature ultrasonic
Transducer 5 can also be customized to ultrasonic transducer specialized factory.
Involved higher-order of oscillation electric signal transmission cable 4 is commercially available;Can also be to ultrasonic transducer producer or cable special manufacturer
Family's customization.
There is the product for being close to needs commercially available on involved higher-order of oscillation electric signal generator market;Can also customize to relevant manufacturers.
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 adnexa or say interface connection of getting lines crossed.
The multi-channel electrochemical work station is commercially available;The multi-channel electrochemical work station can also as needed to specialty
Producer customizes.
In view of its form of this case above related text expresses that it is described pipeline of the presentation parallel construction 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;Immunity is multiple described in this case
Compound refers to the immune complex of broad sense.
Claims (10)
1. the general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled, it is characterised in that the structure of the device includes
Multichannel micro-fluidic chip, the structure of the micro-fluidic chip includes the substrate installed together bonded to each other and cover plate, the substrate
Plate object or tablet are with cover plate, that face towards the cover plate of the substrate is contained via mould pressing process or etching technics shape
Into channel structure, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etching technics
Or the window structure that simple drilling technology is formed, substrate being installed together bonded to each other has been built into and contained jointly with the cover plate
The micro-fluidic chip of pipeline configuration and the liquid pool structure being attached thereto, the locations of structures of the pipeline is located at the substrate and the cover plate phase
The interface zone mutually fitted, 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 described
The locations of structures of liquid pool, the liquid pool has two kinds, two kinds of liquid pools respectively be located at different structure position sample introduction end liquid pool and
Terminal liquid pool, the sample introduction end position of the micro-fluidic chip has one or more sample introduction end liquid pool, and the sample introduction end refers to
Be the micro-fluidic chip actual sample introduction when detected solution injection end position, then have one in the terminal location of the micro-fluidic chip
The terminal liquid pool, the terminal refers to the terminal position of liquid flowing in its chip when the actual sample introduction of the micro-fluidic chip is tested
Put, the terminal is located remotely from each other with the sample introduction end, one end of the pipeline be located at sample introduction end the sample introduction end liquid pool UNICOM, the pipe
The other end in road be located at the micro-fluidic chip the terminal the terminal liquid pool UNICOM, and, sequentially or backward ground point
The working electrode that is not installed in the pipeline on diverse location and to electrode and reference electrode, the order refers to the ginseng
Than its locations of structures of electrode closer to the terminal location, the backward refers to the reference electrode locations of structures closer to described
Sample introduction end position, the working electrode is resisted by conductive electrode and the tumor markerses that embedded being attached on the conductive electrode
The gold size sensitive membrane of body is constituted, and the construction of the pipeline is presented parallel construction, and the pipeline in parallel construction is by 15 branched pipes
Road is in parallel to be constituted, described that the profile that the pipeline of parallel construction its appearance profile is similar to parallel circuit, the work electricity is presented
The quantity of pole is 15, and the installation position of 15 working electrodes is located in 15 laterals respectively, and,
Tumor markerses antibody in the sensitive membrane structure of its top layer gold size of 15 working electrodes is respectively to tumor markerses antigen energy
Specific binding 15 kinds of tumor markerses antibody materials, 15 kinds of antibody materials be respectively tumor markerses antibody A FP,
CEA、CA242、CA125、CA199、CA153、CA724、CA50、NSE、CYFRA21-1、FPSA、TPSA、β-hCG、
SCCA and β 2-MG, the antigen is the antigen of broad sense, and the antibody is the antibody of broad sense, the working electrode its material
It is argent material, gold material, carbon material or thermal decomposition conducting polymer material, the working electrode its pattern is presented post
Shape, lamellar or thread, the substrate its material is polydimethylsiloxane material, and its surface of the substrate is the surface of primary form,
The surface of the primary form its be intended to refer to not through any surface chemical modification or the material of any surface chemical modification
Primary form surface, the structure of the device also includes hinge-type fixture, the hinge-type fixture its appearance profile likeness in form hinge,
The hinge-type fixture is by two hinges being mutually hinged and a fastening screw through two hinges and one
The nut for fastening and hand break loose manually for matching with the fastening screw and connecting together with the fastening screw is constituted, the conjunction
The micro-fluidic chip is respectively mutually drawn close and clamped to two hinges of page fixture with its tip, and the tip is positioned at the micro-fluidic core
In the locations of structures of the neighbouring described terminal of piece, fixation is attached in a hinge at least in and is equiped with miniature ultrasonic
Transducer, and, higher-order of oscillation electric signal transmission cable, one end and the miniature ultrasonic of the higher-order of oscillation electric signal transmission cable
Wave transducer links together;The hinge-type fixture provides a function of facilitating the device to disassemble;The miniature ultrasonic transducing
Device its major function is that, when the actual sample introduction of micro-fluidic chip is tested, the ultrasound wave launched using it is reducing sample solution and institute
State the interfacial tension between the inwall of pipeline so as to can be compatible, also, it is micro- with this using the sample introduction end and the terminal
Difference in the distance between type ultrasonic transducer installation position difference and its ultrasonic intensity for being experienced, induced synthesis
Difference between described its interfacial tension of sample introduction end and the terminal its interfacial tension, the interface between the micro-fluidic chip two ends
Tension difference can form pressure gap between the two ends of the micro-fluidic chip, and the pressure gap can drive sample solution to described
End flow;The miniature ultrasonic transducer units its functions also includes that the ultrasound wave launched with it checks biology contained in sample
Macromole its absorption on the inner surface of pipeline, and then check the substrate of the polydimethylsiloxane material its body phase to the life
The effect of swallowing up of thing macromole;The substrate of the polydimethylsiloxane material its function is included with cover plate and working electrode and to electricity
Pole and reference electrode together build the micro-fluidic chip, soft and have the substrate of the polydimethylsiloxane material of elasticity its function
Also include, with its property to the strong absorption of ultrasound wave, ultrasound wave being absorbed strongly, and thereby at the micro-fluidic chip end
Hold the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end.
2. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In the pipeline is capillary channel including the lateral.
3. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In the thermal decomposition conducting polymer is the conductive material formed Jing after anoxybiotic heat treatment by polyimides or polyacrylonitrile.
4. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the length of the working electrode
Degree is between 1 micron to 15000 microns.
5. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.
6. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In its material of the cover plate in structure is polydimethylsiloxane material.
7. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In the distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.
8. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In, the cover plate in structure and substrate its thickness between 1.0 millimeters and 5.0 millimeters.
9. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
Also include higher-order of oscillation electric signal generator in the structure of, the micro fluidic device, its is another for the higher-order of oscillation electric signal transmission cable
One end is connected with the higher-order of oscillation electric signal generator.
10. general type Diagnostic Value of Several Serum Tumor Markers joint inspection chip apparatus conveniently disassembled according to claim 1, its feature exists
In, the miniature ultrasonic transducer units its specified ultrasonic emitting power between 5 milliwatts and 9000 milliwatts, the miniature ultrasonic
The frequency of wave transducer its ultrasound wave operationally launched is between 100KHz and 12MHz.
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