CN106814184A - The tumor markers joint-detection six-channel microfluidic chip device simplified - Google Patents
The tumor markers joint-detection six-channel microfluidic chip device simplified Download PDFInfo
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Abstract
The present invention relates to a kind of tumor markers joint-detection six-channel microfluidic chip device simplified, belong to analysis testing field.Dimethyl silicone polymer is its substrate for being used for making the multichannel combined detection micro-fluidic chip of classifiable tumor mark of PDMS, advantageous, while there are a series of problems for needing and overcoming；This case is directed to the serial problem.This case main points are, substrate selectes the PDMS with ecosystem surface, and the chain ring type magnetic holding device socket of miniature ultrasonic transducer units will be equiped be positioned at the test liquid stream terminal of the micro-fluidic chip its neighbor positions, with ultrasonic wave reduction interfacial tension, the interface compatibility between related solid-liquid two-phase is significantly increased, while strong absorbabilities of the utilization PDMS to ultrasonic wave, reach ultrasonic intensity rapid decrement in short distance, so as to form interfacial tension difference at the two ends of the chip, test liquid stream is thereby facilitated to be flowed to terminal direction along originally hydrophobic capillary channel.The device exempts to use Micropump completely.
Description
Technical field
The present invention relates to a kind of tumor markers joint-detection six-channel microfluidic chip device simplified, belong to analysis test neck
Domain.
Background technology
Tumor markers (tumor marker, TM) refers in the generation and breeding of tumour, to be produced in itself by tumour cell
It is raw or by body to tumour cell react and produce, reflection tumour exist and growth a class material, including protein,
Hormone, enzyme (isodynamic enzyme) and oncoprotein etc..Tumor markers in chemical examination blood samples of patients or body fluid, can be in cancer screening
Early detection tumour, and observe the curative effect of oncotherapy and judge patient's prognosis.Clinically conventional tumor markers has at present：
(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)
It is 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)
It is 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 swollen ECC, trophoblastic tumor (suede cancer, vesicular mole) etc.
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 for clinically detecting 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 tumor markers exceptions；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 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 and the cover plate that fits together therewith of small fluid course are etched with,
Small fluid course on the substrate, before upper cover plate is assembled, it is apparent on see to be exactly some micro-channels, 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 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 the 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 can be with identical, it is also possible to differ；From material consumption, manufacture difficulty and
Application popularization prospect etc. aspect from the point of view of, 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 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, micro-channel is built on this material, if not carrying out the modified operation for the micro-channel surface, then,
After overall assembling is completed, that is, after covering cover plate, because of structure in the micro-channel its inner surface occupy most liquid circulation
The inner surface in road, then, the PDMS micro-channels inner surface its strong hydrophobic property is deciding factor, and 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
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 by that micro-channel of surface modifying and decorating its inner surface it is modified after state can not maintain foot
Enough long time, being held time for micro-channel its inner surface state after that is surface-modified be substantially only sufficient in completion laboratory
The time of portion's test experiments needs；In other words, PDMS micro-channel inner surfaces of or surface modification modified by surface, its
After modified or say the surface state formed after modification can not be lasting, but soon automatically tend to or say 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, does not do if surface modification, and the fine liquid stream of polar solvent similar to the aqueous solution cannot pump
Send and pass through, chip also cannot just be used；And if having done surface modification, its state after modifying 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 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, the depression that these adsorbed large biological molecules 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, caused by travel motion；Such case, it is also possible to explained from another angle, i.e.
Continuously from inside PDMS body phases to its diffusion into the surface, those polymer molecules of migration, the result of its motion, 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 to be related to the severe deviations of all kinds of test data of experiment of large biological molecule.
As described above, the problem of PDMS substrates is, 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 surface saturation is adsorbed, 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 is explained, there are substantial amounts of Minute pores in PDMS body phases, associated biomolecule macromolecular 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 clamp-ons 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.
In order to prevent this PDMS substrate bodies relative to the effect of swallowing up of large biological molecule, can be from containment PDMS surfaces opposite
The absorption of thing macromolecular is addressed, method be chemically modified aiming at the PDMS material surface it is modified, for
For PDMS is for the situation of substrate material, exactly the surface of described micro-channel part is chemically modified it is modified, 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 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 by the micro-channel inner surface state that surface chemical modification is modified
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
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 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 thorny problem.The problem makes the numerous specialties in this area as another problem addressed above, equally
Personnel are entangled with, perplex 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, by the solution be applied to build it is a kind of it is new can be for six kinds than more typical primary tumor mark
Will thing 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 a kind of tumor markers simplified
Joint-detection six-channel microfluidic chip device, the program is characterized 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 formed via mould pressing process or etching technics, the base in tablet, that face towards the cover plate of the substrate
Piece also contains and be connected with the channel structure and pierce being formed via mould pressing process, etching technics or simple drilling technology for the substrate
Window structure, substrate being installed together bonded to each other 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 respectively the sample introduction end liquid pool and terminal liquid pool for being located at different structure position, 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 the micro-fluidic chip reality
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
Be located remotely from each other, one end of the pipeline and the sample introduction end liquid pool UNICOM for being located at sample introduction end, the other end of the pipeline be located at that this is micro-
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 the reference electrode its locations of structures and more leans 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,
The construction of the pipeline is presented parallel construction, and the pipeline in parallel construction is made up of six lateral parallel connections, and the presentation is simultaneously
The pipeline its appearance profile for joining construction is similar to the profile of parallel circuit, and the quantity of the working electrode is six, this six
The installation position of working electrode is located in six laterals respectively, and, its top layer gold size of six working electrodes is sensitive
Tumor markers antibody in membrane structure is respectively the six kinds of tumor markers antibody that can be specifically bound to tumor markers antigen
Material, six kinds of antibody materials are respectively tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3,
The antigen is the antigen of broad sense, and the antibody is the antibody of broad sense, and the working electrode its material is argent material, gold
Material, carbon material or thermal decomposition conducting polymer material, the working electrode its pattern presentation column, sheet or thread, should
Substrate its material is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, the surface of the primary form its
It is intended to refer to not by any surface chemical modification or the surface of the primary form of the material of any surface chemical modification, should
The structure of device also includes chain ring type magnetic holding device, and six links are had in the chain ring type magnetic holding device its structure, and the link is equal
In strip, shaft-like or bar-shaped, each adjacent link is hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link,
Each drift angle of the hexagonal annular structure its locations of structures is articulated structure position, two therein of the hexagonal annular structure
Link in opposite side position its material is magnetic conductivity material, the remaining four processes of the hexagonal annular structure equal right and wrong of its material
Magnetic material, the non magnetic material refers to the material not responded strongly to externally-applied magnetic field, two rings of the magnetic conductivity material
Magnet exciting coil is wound with a link among section, this is wound with the both sides neighbour position of the link in its magnet exciting coil of magnet exciting coil
The pole shoe for being respectively equiped with a magnetic conductivity material is put, two pole shoes are had, two pole shoes are directed to another magnetic conductivity
The link of material, attaches fixation and is equiped with miniature ultrasonic transducer units on its surface of the link of another magnetic conductivity material, rely on
The chain ring type magnetic holding device socket of electromagnetism adhesive strength is positioned at the close position of its terminal of the micro-fluidic chip, and,
Higher-order of oscillation electric signal transmission cable, one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable are connected to
Together；The chain ring type magnetic holding device provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units its main work(
Can be that, when the actual sample introduction of micro-fluidic chip is tested, the ultrasonic wave launched using it is reduced in sample solution and the pipeline
Interfacial tension between wall, can be compatible, also, is changed with the miniature ultrasonic using the sample introduction end and the terminal
Difference in the distance between energy device installation position difference and its ultrasonic intensity experienced, sample introduction end described in induced synthesis
Difference between its interfacial tension and the terminal its interfacial tension, the interfacial tension difference meeting between the micro-fluidic chip two ends
Pressure gap is formed between the two ends of the micro-fluidic chip, the pressure gap can drive sample solution to the end flow；
The miniature ultrasonic transducer units its functions also include the ultrasonic wave launched with it check contained large biological molecule in sample its
Absorption on the inner surface of pipeline, and then check the substrate of the dimethyl silicone polymer material its body phase to the large biological molecule
The effect of swallowing up；Its function of the substrate of the dimethyl silicone polymer material is including with cover plate and working electrode and to electrode and reference
Electrode together builds the micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function also include with
Its property to the strong absorption of ultrasonic wave, is absorbed strongly to ultrasonic wave, and thereby in the micro-fluidic chip, the terminal is entered to this
The rapid decrement of ultrasonic intensity is realized within limited short distance between sample end.
The tumor markers antibody is the antibody of broad sense, and the antibody of the broad sense is referred to possessing antibody function or is functionally similar to
Can be combined with the various involved tumor markerses of corresponding clinic and forming immune complex or immune compound 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 corresponding
Antibody or be functionally similar to antibody material carry out enzyme mark detection it is various clinical involved the need for differentiate, the tumor-marker of detection
Thing.
The word of the magnet exciting coil one art-recognized meanings of itself are known.
The word of magnetic conductivity material one art-recognized meanings of itself are known；Belong to magnetic conductivity material material such as pure iron,
Silicon steel, soft magnetic ferrite etc..
Non magnetic material described in this case refers to the material not responded strongly to externally-applied magnetic field, belongs to the non magnetic material
Material such as metallic copper, metallic zinc, ormolu, metallic aluminium, almag etc..
The word art-recognized meanings of itself field in Machine Design that is hinged is known.
The gold size sensitive membrane is to be sufficiently mixed uniformly shitosan gold size solution and tumor markers antibody-solutions, uses point sample instrument
Point sample coats specified structure position, and forms its drying and forming-film.Tumor markers antibody in the gold size sensitive membrane is equal
It is the tumor markers antibody that horseradish peroxidase or glucose oxidase are marked, 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 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 aspect such as reagent loss is reduced less as far as possible, described in the pipeline includes
The passage of the preferred capillary level of lateral, the passage of the capillary level implies that the interior of the capillary on its internal diameter and 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
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 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 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 wave transmission powers be between 5 milliwatts and 9000 milliwatts it
Between；The preferred scope of the frequency of the miniature ultrasonic transducer units its ultrasonic waves operationally launched 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
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 required position installed 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 the coating of other appropriate process is quick
Sense film, its thicknesses of layers can allow be any setting the sample measuring liquid treated occur electrical signals response thickness, but, push away
Preferred thickness is between 10 nanometers and 200 nanometers to the thickness recommended in other words conj.or perhaps.
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
Into the micro-fluidic chip of only 2.0 centimetres to 3.0 centimetres of super-small of length, and realized to ultrasonic wave in the extremely short distance
Extremely fast decay, it may be preferred to dimethyl silicone polymer is 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.
The cover plate and substrate its thickness can allow be any setting the thickness for being easy to assembling, 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 application 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 ultrasonic wave, drives liquid
Stream flows in the capillary channel of the six-channel microfluidic chip, using multi-channel electrochemical analyzer device respectively to six kinds of typical cases
Tumor markers antigen carries out joint-detection.
The specific detection of this case micro-fluidic chip is as follows using step：
1st, blood serum sample liquid is added in micro-pipe road, under ultrasonic wave driving, various tumor markers antigen molecules are each
In passage on electrode surface gold size sensitive membrane embedding 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
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 markers antigen.
It is an advantage of the invention that its close position relies on the socket of electromagnetism adhesive strength fixed in the terminal of the micro-fluidic chip
Position described in chain ring type magnetic holding device, with the chain ring type magnetic holding device one magnetic conductivity link attaching installation miniature ultrasonic
Wave transducer, low-power, the ultrasonic wave of high-frequency band launched using it so that without surface chemical modification or come 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 the only several centimeters the sample introduction end
In the very short distance of yardstick, the rapid decrement of ultrasonic intensity is reached, thereby cause institute at the two ends of the micro-fluidic chip
The difference of interfacial tension is stated, and then, using the pressure between its two ends for being formed of the difference of the interfacial tension between the two ends
Difference, drives test liquid stream to be flowed to the terminal direction in such a originally strong hydrophobic capillary channel.By this
Case liquid stream drive scheme, entirely without any coming to the surface must be carried out to the substrate of the dimethyl silicone polymer material and its internal pipeline
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 the Micropump in system meaning etc；On the other hand, the ultrasonic wave of the low-power, high-frequency band, additionally it is possible in containing 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 belonging to the type of 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 that facilitates the Reversible binding between antigen, antibody to react quickly reaches certainly, and this causes correlation
Test operation can be completed with than speed faster.
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 presence, then, the dimethyl silicone polymer
Substrate its body phase interior polymer molecule constantly diffuses to the surface automatically, migrate caused by it to surface chemical modification layer or
The damaging influence of chemically modified layer does not just exist yet.
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, the very cheap polydimethyl siloxane material of this kind is just possible in the micro-fluidic chip
Prepare, production, using etc. field play bigger effect.
Fixation is attached in the chain ring type magnetic holding device one magnetic conductivity link in this case structure and has installed the miniature ultrasonic
Transducer, this architecture provides a function of facilitating the device to disassemble, in this way, the chain ring type magnetic holding device is led together with one
The miniature ultrasonic transducer units attached in magnetic link just easily can be mutually disengaged with the micro-fluidic chip, then, the portion
The component that dividing freely to depart from is reused many times in which just can circulate benignly；The architectural feature is conducive to saving making for the device
Use cost.
The framework of this case micro-fluidic chip its large-scale integrated, determines it among actual test application, to Serum samples
The need for measure smaller, this body and mind for helping to reduce related subject is damaged.
Brief description of the drawings
Fig. 1 be this case device chain ring type magnetic holding device its with outside side view rough under the micro-fluidic chip socket state,
The legend and the relative position relation for indicating under the socket state between each component.
Fig. 2 is the outside side view under another viewing angle of this case device；The viewing angle of Fig. 2 and the sight of Fig. 1
Angle is examined to be mutually perpendicular to.
In figure, 1 is chain ring type magnetic holding device, and 2 is the terminal of the micro-fluidic chip, and 3 is the described of the micro-fluidic chip
Sample introduction end, 4 is the substrate of dimethyl silicone polymer material, and 5 is cover plate, and 6 is miniature ultrasonic transducer units, and 7 is high frequency vibrating
Electric signal transmission cable is swung, 8,15 is respectively two links of magnetic conductivity material for being located at different structure position, two magnetic conductions
Property material link end-to-end relation each other in the structure shown here, 9,22 is respectively two pole shoes for being located at different structure position, 10,
12nd, 14,17,19,21 is respectively that, in six six pin joints of diverse location, 11,13,18,20 is respectively place
In four links of the four of diverse location non magnetic materials, 16 is magnet exciting coil；The chain ring type electromagnetic clamp in legend is signed an undertaking
Structure is only the legend structure illustrated, and the actual chain ring type magnetic holding device structure is not limited to the legend chain ring type magnetic holding device structure；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.
Specific embodiment
In this case that Fig. 1 and Fig. 2 are shown embodiment, the example its main points are that the structure of the device includes that multichannel is micro-
Fluidic chip, the structure of the micro-fluidic chip includes installing bonded to each other substrate 4 and cover plate 5 together, the He of the substrate 4
Cover plate 5 is plate object or tablet, and that face towards the cover plate 5 of the substrate 4 is contained via mould pressing process or etching work
The channel structure that skill is formed, the substrate 4 also containing be connected with the channel structure and pierce the substrate 4 via mould pressing process,
The window structure that etching technics or simple drilling technology are formed, substrate 4 being installed together bonded to each other is common with the cover plate 5
The micro-fluidic chip containing pipeline configuration and the liquid pool structure being attached thereto is built into, the locations of structures of the pipeline is located at the base
Piece 4 and the cover plate 5 interface zone bonded to each other, its side of the window is blocked by the cover plate 5 and opposite side is opened, the window
Locations of structures be exactly the liquid pool locations of structures, the liquid pool has two kinds, and two kinds of liquid pools are respectively to be located at different structure position
There is the one or more sample introduction sample introduction end liquid pool and terminal liquid pool put, the position of sample introduction end 3 of the micro-fluidic chip
End liquid pool, the sample introduction end 3 refers to the injection end position of detected solution during the micro-fluidic chip actual sample introduction, micro-fluidic at this
Then there is a terminal liquid pool position of terminal 2 of chip, and the terminal 2 refers to the micro-fluidic chip actual sample introduction when testing
The terminal location of liquid flowing in its chip, the terminal 2 is located remotely from each other with the sample introduction end 3, one end of the pipeline with positioned at sample introduction
The terminal of the sample introduction end liquid pool UNICOM at end 3, the other end of the pipeline and the terminal 2 positioned at the micro-fluidic chip
Liquid pool UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode with
And reference electrode, the order refers to its locations of structures of the reference electrode closer to the position of the terminal 2, and the backward refers to
Be the reference electrode locations of structures closer to the position of sample introduction end 3, the working electrode is by conductive electrode and attaching
The gold size sensitive membrane for having embedded tumor markers antibody on the conductive electrode is constituted, and the construction of the pipeline is presented structure in parallel
Make, the pipeline in parallel construction is made up of six lateral parallel connections, the described pipeline its profile that parallel construction is presented
In the profile of parallel circuit, the quantity of the working electrode is six to contour approximation, the installation position difference of six working electrodes
In six laterals, and, the tumor markers in the sensitive membrane structure of its top layer gold size of six working electrodes resists
Body is respectively the six kinds of tumor markers antibody materials that can be specifically bound to tumor markers antigen, six kinds of antibody materials difference
It is tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen is the antigen of broad sense, institute
State the antibody that antibody is broad sense, the working electrode its material is that argent material, gold material, carbon material or thermal decomposition are led
Electric macromolecule material, the working electrode its pattern is presented column, sheet or thread, and the substrate 4 its material is poly dimethyl silicon
Oxygen alkane material, its surface of substrate 4 is the surface of primary form, the surface of the primary form its be intended to refer to not by appointing
The surface of the primary form of what surface chemical modification or the material of any surface chemical modification, the structure of the device also includes chain link
Have six links in formula magnetic holding device 1, the chain ring type magnetic holding device 1 its structure, six links be respectively link 8,11,
13rd, 15,18,20, the link 8,11,13,15,18,20 is in strip, shaft-like or bar-shaped, each adjacent link
Be hinged to form chain link, the hexagonal cyclic structure of its appearance profile of the chain link, each drift angle of the hexagonal annular structure its
Locations of structures 10,12,14,17,19,21 is articulated structure position, and two therein of the hexagonal annular structure is in
The link 8,15 of opposite side position its material is magnetic conductivity material, the remaining four processes 11 of the hexagonal annular structure, 13,
18th, 20 its material are non-magnetic materials, and the non magnetic material refers to the material not responded strongly to externally-applied magnetic field,
Magnet exciting coil 16 is wound with a link 15 among two links of the magnetic conductivity material, this is wound with magnet exciting coil 16
Link 15 pole shoe for magnetic conductivity material 9,22 is respectively equiped with the both sides neighbor positions of its magnet exciting coil 16, have
Two pole shoes 9,22, two pole shoes 9,22 are directed to the link 8 of another magnetic conductivity material, another magnetic conduction
Property its surface of the link 8 of material on attach fixation and be equiped with miniature ultrasonic transducer units 6, rely on the chain link of electromagnetism adhesive strength
Formula magnetic holding device 1 is socketed the close position for being positioned at its terminal 2 of the micro-fluidic chip, and, higher-order of oscillation electric signal is passed
Transmission cable 7, one end of the higher-order of oscillation electric signal transmission cable 7 links together with the miniature ultrasonic transducer units 6；The chain
Ring type magnetic holding device 1 provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units 6 its major functions be
When the actual sample introduction of micro-fluidic chip is tested, the ultrasonic wave launched using it is reduced between sample solution and the inwall of the pipeline
Interfacial tension, can be compatible, also, using the sample introduction end 3 and the terminal 2 and the miniature ultrasonic transducing
Difference in the distance between the installation position of device 6 difference and its ultrasonic intensity experienced, sample introduction end described in induced synthesis
Difference between 3 its interfacial tension and the terminal 2 its interfacial tension, the interface between the micro-fluidic chip two ends 2,3
Tension difference can form pressure gap between the two ends 2,3 of the micro-fluidic chip, and the pressure gap can drive sample solution
Flowed to the terminal 2；Its function also ultrasonic wave including being launched with it of miniature ultrasonic transducer units 6 checks institute in sample
The large biological molecule for containing its absorption on the inner surface of pipeline, and then check the substrate 4 of the dimethyl silicone polymer material
Swallow up effect of its body phase to the large biological molecule；The substrate 4 of the dimethyl silicone polymer material its function includes and cover plate 5
And working electrode and the micro-fluidic chip is together built to electrode and reference electrode, soft dimethyl silicone polymer for simultaneously having elasticity
The substrate 4 of material its function also includes with its property to the strong absorption of ultrasonic wave, ultrasonic wave absorbed strongly, and thereby
The fast express delivery of ultrasonic intensity is realized within micro-fluidic chip terminal 2 to the limited short distance between the sample introduction end 3
Subtract.
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 and Fig. 2.
Involved miniature ultrasonic transducer units 6 are commercially available；Can also be customized to ultrasonic transducer producer.
Involved higher-order of oscillation electric signal transmission cable 7 is commercially available；Can also be to ultrasonic transducer producer or cable special manufacturer
Family's customization.
Involved higher-order of oscillation electric signal generator market has the product close to needs commercially available；Can also be customized 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 annex or say interface of getting lines crossed and couple.
The multiple tracks electrochemical workstation is commercially available；The multiple tracks electrochemical workstation can also according to specific needs to related special
Industry producer customizes.
In view of its form of this case related text expresses that it is described above 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；It is immunized described in this case multiple
Compound refers to the immune complex of broad sense.
Claims (10)
1. the tumor markers joint-detection six-channel microfluidic chip device simplified, it is characterised in that the structure bag of the device
Multichannel micro-fluidic chip is included, the structure of the micro-fluidic chip includes installing bonded to each other substrate and cover plate together, the base
Piece and cover plate are plate object or tablet, and that face towards the cover plate of the substrate is contained via mould pressing process or etching technics
The channel structure of formation, the substrate also containing be connected with the channel structure and pierce the substrate via mould pressing process, etch work
The window structure that skill or simple drilling technology are formed, substrate being installed together bonded to each other has been built into and has contained jointly with the cover plate
The locations of structures for having the micro-fluidic chip of pipeline configuration and the liquid pool structure being attached thereto, the pipeline is located at the substrate and the 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 institute
State 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 with
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
The injection end position of detected solution during the micro-fluidic chip actual sample introduction is referred to, then has one in the terminal location of the micro-fluidic chip
The individual 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 markers 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 six laterals
Parallel connection is constituted, and the described pipeline its appearance profile that parallel construction is presented is similar to the profile of parallel circuit, the working electrode
Quantity be six, six installation positions of working electrode respectively be located at six laterals in, and, six works
The tumor markers antibody made in electrode sensitive membrane structure of its top layer gold size is respectively that tumor markers antigen can be specifically bound
Six kinds of tumor markers antibody materials, six kinds of antibody materials be respectively tumor markers antibody A FP, CEA, PSA, CA125,
CA19-9 and CA15-3, 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 presentation post
Shape, sheet or thread, the substrate its material is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form,
Its material for being intended to refer to not by any surface chemical modification or any surface chemical modification of the surface of the primary form
Primary form surface, the structure of the device also includes chain ring type magnetic holding device, is had in the chain ring type magnetic holding device its structure
Six links, in strip, shaft-like or bar-shaped, each adjacent link is hinged to form chain link the link, the chain link its profile
The hexagonal cyclic structure of profile, each drift angle of the hexagonal annular structure its locations of structures is articulated structure position, and this six
Two therein of side shape cyclic structure is magnetic conductivity material in the link of opposite side position its material, the hexagonal annular structure it is remaining
Lower its material of four processes is non-magnetic material, and the non magnetic material refers to the material not responded strongly to externally-applied magnetic field
Matter, magnet exciting coil is wound with a link among two links of the magnetic conductivity material, and this is wound with the link of magnet exciting coil
A pole shoe for magnetic conductivity material is respectively equiped with the both sides neighbor positions of its magnet exciting coil, two pole shoes are had, this two
Individual pole shoe is directed to the link of another magnetic conductivity material, and fixed installing is attached on its surface of the link of another magnetic conductivity material
There are miniature ultrasonic transducer units, rely on the chain ring type magnetic holding device socket of electromagnetism adhesive strength to be positioned at its institute of the micro-fluidic chip
State the close position of terminal, and, higher-order of oscillation electric signal transmission cable, one end of the higher-order of oscillation electric signal transmission cable with
The miniature ultrasonic transducer units link together；The chain ring type magnetic holding device provides a function of facilitating the device to disassemble；Should
Miniature ultrasonic transducer units its major functions is that, when the actual sample introduction of micro-fluidic chip is tested, the ultrasonic wave launched using it is dropped
Interfacial tension between the inwall of low sample solution and the pipeline, can be compatible, also, using the sample introduction end and
In the distance between the terminal and the miniature ultrasonic transducer units installation position difference and its ultrasonic intensity experienced
Difference, the difference between its interfacial tension of sample introduction end described in induced synthesis and the terminal its interfacial tension, the micro-fluidic chip
Interfacial tension difference between the two ends can form pressure gap between the two ends of the micro-fluidic chip, and the pressure gap can drive
Dynamic sample solution is to the end flow；The miniature ultrasonic transducer units its function also ultrasonic wave including being launched with it checks examination
Its absorption on the inner surface of pipeline of contained large biological molecule in sample, and then check the dimethyl silicone polymer material
Swallow up effect of its body phase of substrate to the large biological molecule；The substrate of the dimethyl silicone polymer material its function includes and cover plate
And working electrode and the micro-fluidic chip is together built to electrode and reference electrode, soft dimethyl silicone polymer for simultaneously having elasticity
The substrate of material its function also includes with its property to the strong absorption of ultrasonic wave, ultrasonic wave is absorbed strongly, and thereby exist
The micro-fluidic chip terminal is to the rapid decrement that ultrasonic intensity is realized within the limited short distance between the sample introduction end.
2. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the pipeline is capillary channel including the lateral.
3. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the thermal decomposition conducting polymer is the conductive material formed after being heat-treated through anoxybiotic by polyimides or polyacrylonitrile.
4. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
Be, the width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the working electrode
Length is between 1 micron to 15000 microns.
5. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.
6. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that its material of the cover plate in structure is dimethyl silicone polymer material.
7. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.
8. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the cover plate in structure and substrate its thickness are between 1.0 millimeters and 5.0 millimeters.
9. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
Be that 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
The other end is connected with the higher-order of oscillation electric signal generator.
10. the tumor markers joint-detection six-channel microfluidic chip device simplified according to claim 1, its feature
It is that the miniature ultrasonic transducer units its specified ultrasonic wave transmission powers is between 5 milliwatts and 9000 milliwatts, and this is miniature super
The frequency of acoustic wave transducer its ultrasonic wave operationally launched is between 100KHz and 12MHz.
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| CN107655879A (en) * | 2017-09-01 | 2018-02-02 | 北京华科泰生物技术有限公司 | For the micro-fluidic chemiluminescence detection system for the magnetic particle for detecting sexual gland series |
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