CN106563513A - Microfluidic chip device for combined detection of six typical tumor markers - Google Patents
Microfluidic chip device for combined detection of six typical tumor markers Download PDFInfo
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Abstract
Belonging to the field of analytical test, the invention realtes to a microfluidic chip device for combined detection of six typical tumor markers. Use of polydimethylsiloxane, i.e. PDMS as a substrate of the microfluidic chip for multichannel combined detection of typical tumor markers has advantages and problems. The invention aims to solve the problems. The key points of the invention lie in that: PDMS with an original ecological surface is selected as the substrate, holding arms of the substrate are attached with an elastic clip of a miniature ultrasonic transducer to achieve elastic occlusion positioning on a sample liquid flow terminal neighbouring position of the microfluidic chip, the interfacial tension is reduced by ultrasonic wave, thus greatly increasing the interfacial compatibility between a solid phase and a liquid phase, at the same time the strong absorption capacity of PDMS to ultrasonic wave is utilized to achieve rapid diminishing of ultrasonic wave intensity in a short distance, thus forming interfacial tension difference at two ends of the chip, and promoting the flow of the sample liquid flow to the terminal direction along an original strongly hydrophobic capillary channel. The device is free of micropump.
Description
Technical field
The present invention relates to a kind of six kinds of tumor markers joint-detection micro flow control chip devices of typical case, belong to analysis field tests.
Background technology
Tumor markers (tumor marker, TM) is referred in the generation and breeding of tumour, produced by tumour cell itself
It is raw or by body to tumour cell react and produce, reflect tumour exist and grow 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.At present clinically conventional tumor markers has：
(1) alpha-fetoprotein (AFP) is the mark of the tumours such as primary carcinoma of liver, carcinoma of testis, oophoroma；(2) carcinomebryonic antigen (CEA) is digestion
The mark of the tumours such as system tumor, lung cancer, breast cancer；(3) CA125 (CA125) is the mark of the tumours such as oophoroma；
(4) CA153 (CA153) is the mark of the tumours such as breast cancer；(5) CA19-9 (CA19-9) is digestive system tumor
Mark；(6) CA724 (CA724) is the mark of the tumours such as cancer of the stomach, oophoroma；(7) carbohydrate antigen 242 (CA242)
For the mark of digestive system tumor；(8) CA50 (CA50) is the mark of the tumours such as digestive system tumor, breast cancer, lung cancer
Thing；(9) CYFRA21-1 (Cy211) is the mark of the tumours such as non-small cell lung cancer；(10) neuronspecific enolase (NSE)
For the mark of the tumours such as ED-SCLC, neuroendocrine tumor；(11) PSA (PSA) is prostate cancer
Tumor markers；(12) human chorionic gonadotrophin (HCG) is that ECC, trophoblastic tumor (suede cancer, vesicular mole) etc. are swollen
The mark of knurl；(13) thyroglobulin (TG) is the mark of thyroid cancer；(14) ferritin (SF) is digestive system tumor, liver
The mark of the tumours such as cancer, breast cancer, lung cancer；(15) B2M (β 2-MG) is in chronic lymphocytic leukemia, lymph
Raise 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 occurs in breast cancer；CEA often occurs in intestinal cancer, cancer of the stomach；CA19-9 is often occurred in
Intestinal cancer, cancer of pancreas；CA125 often occurs 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 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-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 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, quartz plate, sheet glass, high polymer such as dimethyl silicone polymer, 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, comparatively dimethyl silicone polymer, i.e. PDMS are very easily molded, 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-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 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 surfaces of or surface modification modified through 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 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 the 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 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, 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 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 impact 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.
With regard to 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, as dependence test the biology of object is analyzed
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 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 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 through 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 the 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 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, 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 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 six kinds of tumor-markers of typical case
Thing joint-detection micro flow control chip device, the program is characterized in 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 markers 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 six lateral parallel connections, described to be presented in parallel
The pipeline its appearance profile of construction is similar to the profile of parallel circuit, and the quantity of the working electrode is six, six works
Make the installation position of electrode to be located in six laterals respectively, and, six working electrodes its top layer gold size sensitive membranes
Tumor markers antibody in structure is respectively the six kinds of tumor markers antibody things that can be specifically bound to tumor markers antigen
Matter, six kinds of antibody materials are respectively tumor markers antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, institute
State the antigen that antigen is broad sense, the antibody is the antibody of broad sense, the working electrode its material is argent material, gold material
Matter, carbon material or thermal decomposition conducting polymer material, the working electrode its pattern is presented column, sheet or thread, the base
Piece its material is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, its meaning of the surface of the primary form
Think of is referred to not through any surface chemical modification or the surface of the primary form of the material of any surface chemical modification, the dress
The structure put also includes elastic clip, and two opposite clamping limb snap-in locations of the elastic clip are in the neighbouring described of the micro-fluidic chip
The position of terminal, attaches fixation and is equiped with miniature ultrasonic transducer units on a clamping limb at least in, and, it is high
Frequency vibration swings electric signal transmission cable, and one end and the miniature ultrasonic transducer units of the higher-order of oscillation electric signal transmission cable are connected to one
Rise；The elastic clip provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units its major functions is in miniflow
During the actual sample introduction test of control chip, the ultrasonic wave launched using it is reducing the boundary between sample solution and the inwall of the pipeline
Face tension force so as to can be compatible, also, using the sample introduction end and the terminal and the miniature ultrasonic transducer units installing position
The distance between put difference and the difference on its ultrasonic intensity for being experienced, its interfacial tension of sample introduction end described in induced synthesis
With the difference between the terminal its interfacial tension, the interfacial tension difference between the micro-fluidic chip two ends can be micro-fluidic at this
Pressure gap is formed between the two ends of chip, the pressure gap can drive sample solution to the end flow；The miniature ultrasonic
Wave transducer its function also include the ultrasonic wave launched with it check large biological molecule contained in sample its in the pipeline
Absorption on inner surface, and then check swallow up work of its body phase of the substrate of the dimethyl silicone polymer material to the large biological molecule
With；The substrate of the dimethyl silicone polymer material its function is included with cover plate and working electrode and to electrode and reference electrode together
Build the micro-fluidic chip, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include with it to ultrasound
The property that ripple absorbs strongly, is absorbed strongly to ultrasonic wave, and thereby in the micro-fluidic chip terminal between the sample introduction end
Limited short distance within realize the rapid decrement of ultrasonic intensity.
The tumor markers 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 markers of various corresponding clinics
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 clinic involved by the tumor-marker for needing to differentiate, detect
Thing.
Described miniature ultrasonic transducer units can certainly be all installed on two clamping limbs；But, only installing one is miniature
Ultrasonic transducer is dealt with enough and is used.
The word of elastic clip one art-recognized meanings of itself are known.
The gold size sensitive membrane is to be sufficiently mixed shitosan gold size solution and tumor markers antibody-solutions uniformly, 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
For the tumor markers antibody that horseradish peroxidase or glucose oxidase are marked, the gold size sensitive membrane is 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 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 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 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, sheet, 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 wave transmission powers be between 5 milliwatts and 5000 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 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, 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 realize 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, select 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 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 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 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 ultrasonic wave drives, 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 immunity again
Compound.
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 markers antigen.
It is an advantage of the invention that in its elastic clip described in close position snap-in location of 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 on the clamping limb of the elastic clip
Ultrasonic wave so that without its pipe of strong hydrophobic micro-fluidic chip internal pipeline of surface chemical modification or surface chemical modification
Compatibility between wall and the test object aqueous solution is significantly increased, and this is test liquid stream by there is provided a realistic possibility；
Meanwhile, using its strong absorbability to ultrasonic wave of dimethyl silicone polymer substrate, in shorter distance, it is,
In from the terminal to the very short distance of the only several centimeters yardstick the sample introduction end, the fast express delivery of ultrasonic intensity is reached
Subtract, the difference of the interfacial tension is thereby caused at the two ends of the micro-fluidic chip, and then, using the boundary between the two ends
Pressure gap between its two ends for being formed of the difference of face tension force, drives test liquid stream originally strong hydrophobic in such a
To terminal direction flowing in capillary channel.By this case liquid stream drive scheme, entirely without must be to the dimethyl silicone polymer
The substrate and its internal pipeline of material carries out any surface chemical modification or chemical modification, has altogether dispensed with the surface chemical modification
Or the laborious procedures of chemical modification；And altogether dispense with the equipment of traditional Micropump etc；On the other hand, the low work(
The ultrasonic wave of rate, high-frequency band, additionally it is possible to contain the large biological molecule in sample in the literalness naked dimethyl silicone polymer
Absorption on substrate its inner surface of pipeline, and then contain that the dimethyl silicone polymer substrate its body phase is gulped down to the large biological molecule
Do not act on；The Reversible binding thing of the antigen, antibody and antigen and antibody is all belonging to the class of described large biological molecule certainly
Type；Because described suction-operated and the described effect of swallowing up effectively are contained, therefore, dependence test result more will can
It is enough objectively to reflect actual conditions；The effect of the low-power, high-frequency band ultrasonic wave, certainly also include facilitate antigen, antibody it
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 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 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 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
Preparation, production, application etc. field plays bigger effect.
The miniature ultrasonic transducer units have been installed in fixation on the elastic clip its clamping limb in this case structure, be this architecture provides
One facilitates the function that the device is disassembled, thus, the elastic clip just can be conveniently together with miniature ultrasonic transducer units appended thereon
Ground is mutually disengaged with the micro-fluidic chip, then, the component that the part can freely depart from is reused and permitted in which just can circulate benignly
Repeatedly；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 elastic clip, and 2,7 is respectively two clamping limbs of elastic clip, and 3 is the terminal of the micro-fluidic chip,
4 is the sample introduction end of the micro-fluidic chip, and 5 is the substrate of dimethyl silicone polymer material, and 6 is cover plate, and 8 is miniature super
Acoustic wave transducer, 9 is higher-order of oscillation electric signal transmission cable, and 10 is tension spring；The spring clamp structure in legend only shows
The legend structure of meaning, actual spring clamp structure is not limited to the legend spring clamp structure；It is micro-fluidic that arrow in legend indicates this
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 5 installed together bonded to each other and cover plate 6, the substrate 5 and cover plate 6
Plate object or tablet are, that face towards the cover plate 6 of the substrate 5 is contained and formed via mould pressing process or etching technics
Channel structure, the substrate 5 also containing be connected with the channel structure and pierce the substrate 5 via mould pressing process, etch work
The window structure that skill or simple drilling technology are formed, substrate 5 being installed together bonded to each other is built into jointly with the cover plate 6
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 5 with
The cover plate 6 interface zone bonded to each other, its side of the window is blocked by the cover plate 6 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 4 of the micro-fluidic chip
Pond, the sample introduction end 4 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 3 then have a terminal liquid pool, the terminal 3 refers to its core when the actual sample introduction of the micro-fluidic chip is tested
The terminal location of liquid flowing in piece, the terminal 3 is located remotely from each other with the sample introduction end 4, one end of the pipeline with positioned at sample introduction end 4
The sample introduction end liquid pool UNICOM, the other end of the pipeline be located at the micro-fluidic chip the terminal 3 the terminal liquid pool
UNICOM, and, sequentially or backward be respectively installed in working electrode in the pipeline on diverse location and to electrode and ginseng
Than electrode, the order refers to its locations of structures of the reference electrode closer to the position of the terminal 3, and the backward is referred to
, closer to the position of sample introduction end 4, the working electrode is by conductive electrode and is attached to this for the reference electrode locations of structures
The gold size sensitive membrane for having embedded tumor markers antibody on conductive electrode is constituted, and the construction of the pipeline is presented parallel construction, institute
State the pipeline in parallel construction to be made up of six lateral parallel connections, its appearance profile of the pipeline of the presentation parallel construction is near
The profile of parallel circuit is similar to, the quantity of the working electrode is six, the installation position of six working electrodes is located at respectively institute
State in six laterals, and, the tumor markers antibody difference in the sensitive membrane structure of its top layer gold size of six working electrodes
It is six kinds of tumor markers antibody materials that tumor markers antigen can be specifically bound, six kinds of antibody materials are respectively tumours
Mark antibody A FP, CEA, PSA, CA125, CA19-9 and CA15-3, the antigen is the antigen of broad sense, the antibody
It is the antibody of broad sense, the working electrode its material is argent material, gold material, carbon material or thermal decomposition conductive polymer
Sub- material, the working electrode its pattern is presented column, sheet or thread, and the substrate 5 its material is dimethyl silicone polymer material
Matter, its surface of substrate 5 is the surface of primary form, the surface of the primary form its be intended to refer to not through any surface
The surface of the primary form of the material of chemical modification or any surface chemical modification, the structure of the device also includes elastic clip 1,
Two opposite clamping limbs 2,7 snap-in locations of the elastic clip 1 in the position of the neighbouring described terminal 3 of the micro-fluidic chip,
A clamping limb at least in for example attaches fixation on clamping limb 7 and is equiped with miniature ultrasonic transducer units 8, and,
Higher-order of oscillation electric signal transmission cable 9, one end of the higher-order of oscillation electric signal transmission cable 9 and the miniature ultrasonic transducer units 8
Link together；The elastic clip 1 provides a function of facilitating the device to disassemble；Its is main for the miniature ultrasonic transducer units 8
Function is that, when the actual sample introduction of micro-fluidic chip is tested, the ultrasonic wave launched using it is reducing sample solution with the pipeline
Interfacial tension between inwall so as to can be compatible, also, it is miniature super with this using the sample introduction end 4 and the terminal 3
Difference in the distance between the installation position of acoustic wave transducer 8 difference and its ultrasonic intensity for being experienced, induced synthesis institute
State the difference between its interfacial tension of sample introduction end 4 and the terminal 3 its interfacial tension, the micro-fluidic chip two ends 3,4 it
Between interfacial tension difference can form pressure gap between the two ends 3,4 of the micro-fluidic chip, the pressure gap can drive
Sample solution to the terminal 3 flows；The miniature ultrasonic transducer units 8 its functions also includes that the ultrasonic wave launched with it is checked
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 5 to the large biological molecule；The substrate 5 of the dimethyl silicone polymer material its function includes
The micro-fluidic chip, the poly- diformazan that is soft and having elasticity are together built with cover plate 6 and working electrode and to electrode and reference electrode
The substrate 5 of radical siloxane material its function also includes, with its property to the strong absorption of ultrasonic wave, ultrasonic wave being inhaled strongly
Receive, and thereby within micro-fluidic chip terminal 3 to the limited short distance between the sample introduction end 4 realize ultrasonic intensity
Rapid decrement.
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.
Itself is commercially available for involved elastic clip.It is available for commercially available alternative spring clamp structure and make that this case structure uses
Type and size etc., quality is various, and concrete elastic clip pattern can be selected as needed.
Involved miniature ultrasonic transducer units 8 are commercially available；Can also customize to ultrasonic transducer producer.
Involved higher-order of oscillation electric signal transmission cable 9 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 for being close to needs commercially available；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 annex or say interface connection of getting lines crossed.
The multiple tracks electrochemical workstation is commercially available；The multiple tracks electrochemical workstation can also be special to correlation according to specific needs
Industry 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. typical six kinds of tumor markers joint-detection micro flow control chip devices, it is characterised in that the structure of the device includes many
Channel microfluidic chip, the structure of the micro-fluidic chip includes installing bonded to each other substrate together and cover plate, the substrate and
Cover plate is plate object or tablet, and that face towards the cover plate of the substrate is contained and formed via mould pressing process or etching technics
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 containing pipe jointly with the cover plate
The micro-fluidic chip of road structure and the liquid pool structure being attached thereto, the locations of structures of the pipeline is mutual with the cover plate positioned at the substrate
The interface zone of laminating, 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 liquid
The locations of structures in pond, the liquid pool has two kinds, and two kinds of liquid pools are respectively sample introduction end liquid pool and the ends for being located at different structure position
End liquid pool, the sample introduction end position of the micro-fluidic chip has one or more sample introduction end liquid pool, what the sample introduction end referred to
The injection end position of detected solution, then has an institute in the terminal location of the micro-fluidic chip when being the micro-fluidic chip actual sample introduction
Terminal liquid pool is stated, the 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 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 pipeline
The other end be located at the micro-fluidic chip the terminal the terminal liquid pool UNICOM, and, sequentially or backward ground fill respectively
The working electrode that is located in the pipeline on diverse location and to electrode and reference electrode, the order refers to the reference electricity
Closer to the terminal location, the backward refers to the reference electrode locations of structures closer to the sample introduction to extremely locations of structures
End position, the working electrode is by conductive electrode and is attached on the conductive electrode and has embedded tumor markers antibody
Gold size sensitive membrane is constituted, and the construction of the pipeline is presented parallel construction, and the pipeline in parallel construction is by six lateral parallel connections
Constitute, the described pipeline its appearance profile that parallel construction is presented is similar to the profile of parallel circuit, the number of the working electrode
Amount is six, and the installation position of six working electrodes is located in six laterals respectively, and, this six work electricity
Tumor markers antibody extremely in top layer gold size sensitivity membrane structure is respectively six can specifically bound to tumor markers antigen
Tumor markers antibody materials are planted, 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 is presented column, sheet
Or it is thread, the substrate its material is dimethyl silicone polymer material, and its surface of the substrate is the surface of primary form, the primary shape
The surface of state its be intended to refer to not through any surface chemical modification or the primary shape of the material of any surface chemical modification
The surface of state, the structure of the device also includes elastic clip, and two opposite clamping limb snap-in locations of the elastic clip are micro-fluidic at this
The position of the neighbouring described terminal of chip, attaching fixation on a clamping limb at least in is equiped with miniature ultrasonic and changes
Energy device, 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
Transducer links together；The elastic clip provides a function of facilitating the device to disassemble；The miniature ultrasonic transducer units its master
Function is wanted to be that, when the actual sample introduction of micro-fluidic chip is tested, the ultrasonic wave launched using it is reducing sample solution with the pipeline
Inwall between interfacial tension so as to can be compatible, also, using the sample introduction end and the terminal and the miniature ultrasonic
Difference in the distance between wave transducer installation position difference and its ultrasonic intensity for being experienced, enters described in induced synthesis
Difference between its interfacial tension of sample end and the terminal its interfacial tension, the interfacial tension between the micro-fluidic chip two ends is poor
Different to form pressure gap between the two ends of the micro-fluidic chip, the pressure gap can drive sample solution to the terminal stream
It is dynamic；The miniature ultrasonic transducer units its functions also includes that the ultrasonic wave launched with it checks large biological molecule contained in sample
Its absorption on the inner surface of pipeline, and then check its body phase of the substrate of the dimethyl silicone polymer material to the biology greatly point
The effect of swallowing up of son；The substrate of the dimethyl silicone polymer material its function is included with cover plate and working electrode and to electrode and ginseng
The micro-fluidic chip is together built than electrode, it is soft and have the substrate of the dimethyl silicone polymer material of elasticity its function and also include
With its property to the strong absorption of ultrasonic wave, ultrasonic wave is absorbed strongly, and thereby in the micro-fluidic chip terminal to this
The rapid decrement of ultrasonic intensity is realized within limited short distance between sample introduction end.
2. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The pipeline is capillary channel including the lateral.
3. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The thermal decomposition conducting polymer is the conductive material formed Jing after anoxybiotic heat treatment by polyimides or polyacrylonitrile.
4. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The width or diameter of the working electrode between 0.1 micron to 2000 microns, and, the length of the working electrode is 1
Micron is between 15000 microns.
5. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The thickness of the gold size sensitive membrane is between 10 nanometers and 200 nanometers.
6. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
Its material of the cover plate in structure is dimethyl silicone polymer material.
7. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The distance between the terminal and the sample introduction end are between 3 centimetres and 10 centimetres.
8. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
The cover plate in structure and substrate its thickness are between 1.0 millimeters and 5.0 millimeters.
9. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in 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 other end
It is connected with the higher-order of oscillation electric signal generator.
10. typical six kinds of tumor markerses joint-detection micro flow control chip device according to claim 1, it is characterised in that
Between 5 milliwatts and 5000 milliwatts, the miniature ultrasonic is changed for the miniature ultrasonic transducer units its specified ultrasonic wave transmission powers
Can the frequency of device its ultrasonic wave operationally launched be between 100KHz and 12MHz.
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|---|---|---|---|
| CN201510686539.1A CN106563513A (en) | 2015-10-12 | 2015-10-12 | Microfluidic chip device for combined detection of six typical tumor markers |
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|---|---|---|---|
| CN201510686539.1A CN106563513A (en) | 2015-10-12 | 2015-10-12 | Microfluidic chip device for combined detection of six typical tumor markers |
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| US20090242059A1 (en) * | 2005-09-28 | 2009-10-01 | Emil Kartalov | Pdms microfluidic components and methods of operation of the same |
| CN101587124A (en) * | 2009-07-12 | 2009-11-25 | 宁波大学 | Micro-fluidic chip special for diagnosing syphilis by aid of organic conductor material technology |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090242059A1 (en) * | 2005-09-28 | 2009-10-01 | Emil Kartalov | Pdms microfluidic components and methods of operation of the same |
| CN101587124A (en) * | 2009-07-12 | 2009-11-25 | 宁波大学 | Micro-fluidic chip special for diagnosing syphilis by aid of organic conductor material technology |
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