CN202083692U - Microfluidic chip specially used for gastric cancer diagnosis - Google Patents
Microfluidic chip specially used for gastric cancer diagnosis Download PDFInfo
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- CN202083692U CN202083692U CN2011200826263U CN201120082626U CN202083692U CN 202083692 U CN202083692 U CN 202083692U CN 2011200826263 U CN2011200826263 U CN 2011200826263U CN 201120082626 U CN201120082626 U CN 201120082626U CN 202083692 U CN202083692 U CN 202083692U
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- gastric cancer
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- diagnosing gastric
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Abstract
The utility model relates to a gastric cancer diagnosis device with a composite nanometer particulate modified electrode, in particular to a microfluidic chip which belongs to the field of analysis and test. The inside of the microfluidic chip comprises pipelines in a parallel structure; the parallel structure comprises four branch pipelines which are connected in parallel; four working electrodes are respectively assembled in the four branch pipelines; the working electrodes consist of metal electrodes and mixtures coated on the surfaces of the working electrodes and comprising graphene, nanogold, gastric cancer diagnosis antibodies and macromolecular Nafion liquid; the gastric cancer diagnosis antibodies on the surfaces of the four working electrodes are respectively specific antibodies comprising CEA antibody, CA12-5 antibody, CA19-9 antibody and CA72-4 antibody. The microfluidic chip has a simple and integrated structure. The microfluidic chip is beneficial for quickening the gastric cancer diagnosis, improving the diagnosis accuracy and reducing the diagnosis cost.
Description
[technical field]
The utility model relates to a kind of diagnosing gastric cancer device of composite nano particle modified electrode, and this diagnosing gastric cancer device is based on the micro-fluidic chip special that the antigen/antibody specific reaction comes diagnosis of gastric cancer, belongs to the analytical test field.
[background technology]
Cancer of the stomach (Stomach cancer) is one of modal cancer of digestive system, occupy disease of digestive tract cause of death first place, China's gastric cancer mortality is the 25.2/10 ten thousand (male sex: 32.8/10 ten thousand, women: 17.0/10 ten thousand), account for 23.2% of whole cancer mortalities, occupy first (male sex is 1.9 times of women) of cancer mortality.Therefore, exploitation is simple, fast, hypersensitivity and specificity diagnostic method, reagent and device, it is the effective way of preventing and treating cancer of the stomach and reducing case fatality rate that cancer of the stomach is carried out early diagnosis and early treatment.
At present, being used for the method for diagnosis of gastric cancer clinically has: fiberendoscopy, exfoliative cytology inspection, ultrasound diagnosis, CT examination, X-ray examination and methods such as biology and biochemical analysis.Though the first five kind method detection technique is accurately and reliably, apparatus expensive, pre-treatment is loaded down with trivial details consuming time and need the laboratory and the medical personnel of specialty, is difficult to promote also be popularized for on-the-spot and cheap application of vast patients with gastric cancer.Have the distinctive high selectivity of immunoassay itself based on immunologic biology and biochemical analysis method,, judge that aspects such as curative effect and prognosis are all significant the early diagnosis of cancer of the stomach.Yet the detection more complicated of cancer of the stomach, traditional immunoassay only detects at its single diagnostic antigen usually.Owing to may appear in the different cancers with a kind of diagnostic antigen, or diagnostic antigen not of the same race can occur in same cancer simultaneously, the diagnostic antigen of cancer of the stomach mainly contains: CEA antigen, CA12-5 antigen, CA19-9 antigen, several antigens such as CA72-4 antigen, therefore, carry out multiple efficiently immune joint inspection analytical approach, simultaneously qualitative truly from the angle of biological correlativity, cancer of the stomach is prevented and detected to the kind of the multiple diagnostic antigen of quantitative response and quantity in early days, judge the cancer of the stomach development degree, aspects such as O﹠A curing gastric cancer effect all have important medical and are worth.
Electrochemical immunosensor combines the high sensitivity of Electrochemical Detection and the high specific of immunoassay, have outstanding advantages such as amount of samples is little, detection is quick, the result is accurate, be widely used in the cancer diagnosis detection of antigens, unique advantages such as its automaticity height, cheapness, easily preparation, rapid sensitive, cooperate immunoreactive efficiently single-mindedly, be fit to make up very much the analyser that the multiple immunoassay of cancer of the stomach detects.The micro-fluidic chip that the title of " life science integrated circuit " is arranged is the desirable technique that makes up microminiaturized, integrated electrochemical immunosensor.The micro-fluidic chip technology be with sampling, pre-service, add reagent, reaction, separation, detection etc. be integrated in finish on the microchip one in front of the door along technology, have analysis speed fast, contain much information, reagent consumption less, advantage and very suitable suitability for industrialized production such as pollutions is little, operation cost is low, instrument is easy to use.On behalf of the 21 century analytical instrument, microflow control technique move towards microminiaturized, integrated developing direction.The micro-fluidic Amperometric Detection Coupled chip that the cancer of the stomach that is suitable for structure detects has following advantage: (1) is easy to integrated a plurality of sense channels and detecting electrode on chip by micro-processing technology, accomplishes that multiple diagnosing gastric cancer antigen analyzes the technical feasibility height simultaneously.(2) by the micro-flowing injection technology, mobile the guaranteed electrode surface of sample in pipeline upgrades constantly, overcome better that electrode is easily contaminated, false positive rate is high, problem such as collimation difference as a result.(3) microfluidic sensing device upper pipeline diameter has only μ m level, and the reaction tank volume also has only μ L level, and required reagent dosage is few during analysis, and analyte arrives the diffusion length of electrode surface lacks, therefore can significantly reduce the incubation time, realizes the fast joint inspection target.(4) the microfluidic sensing device is many based on the optical detection system at present, and the expensive volume of instrument is big, fully integrated being difficult to, and also it is very high to detect cost, has limited its popularization.And adopting Electrochemical Detection in the ampere immunity sensor, volume is little, automaticity is high, cost is low.
At present, in the micro-fluidic chip technical field, detect the correlation technique of quick diagnosis cancer of the stomach and method not to appear in the newspapers as yet when utilizing the multiple diagnostic antigen of cancer of the stomach.
[utility model content]
Technical problem to be solved in the utility model is, in the so total technological frame of micro-fluidic chip technology, develop a kind of multiple diagnosis antibody that can utilize cancer of the stomach and come that multiple diagnostic antigen to cancer of the stomach detects simultaneously, the micro-fluidic chip special of quick diagnosis.
The utility model solve the technical problem by following scheme, the device that this scheme provides is a kind of micro-fluidic chip of diagnosing gastric cancer special use, the structure of this micro-fluidic chip comprises that applying is installed in two plate objects together, described two plate objects are respectively a cover plate and a substrate, on the position of the mutual applying between described two plate objects, be equiped with pipeline, three pond shape things and be installed in the working electrode on the diverse location in the described pipeline in regular turn respectively, to electrode and contrast electrode, one end of pipeline via the manifold shape fluid passage respectively with wherein two pond shape thing UNICOMs, the other end of pipeline and another remaining pond shape thing UNICOM, described working electrode is made up of metal electrode and the film that is coated in surface of metal electrode, and described film is by Graphene, nm of gold, the potpourri of diagnosing gastric cancer antibody and macromolecule Nafion solution constitutes.
As further improvement of the utility model, the structure of described pipeline is structure in parallel, the described pipeline that is structure in parallel is made of four lateral parallel connections, and, the quantity of described working electrode is four, the installation position of these four working electrodes lays respectively in described four laterals, and the diagnosing gastric cancer antibody materials that these four working electrode top layers apply is respectively CEA antibody, CA12-5 antibody, CA19-9 antibody and the CA72-4 antibody that can combine with diagnosing gastric cancer antigen energy specificity.
Film on the described working electrode is the composite nano particle sensitive membrane, wherein embedding diagnosing gastric cancer antibody.This film is Graphene, nm of gold, various diagnosing gastric cancer antibody are fully mixed with high molecular Nafion solution and to vibrate evenly, drips in surface of metal electrode and its dried overnight film forming is formed by micro syringe.
Pipeline in the described microfluidic chip structure, its internal diameter size all can be selected arbitrarily size, but, for as far as possible less with the consideration of aspects such as liquid sample to be measured and reduction reagent loss, described pipeline is preferably all selected the passage of capillary level for use, the passage meaning of described capillary level is a capillary channel, and the internal diameter capillaceous on its internal diameter and the ordinary meaning is suitable.The shape of cross section of its inner passage of described kapillary can be a shape arbitrarily, described shape of cross section is for example circular, oval, square, rectangle, bar shaped, can certainly be have bending arbitrarily linear, and, described interior shape capillaceous is along with the extension of pipeline, and it is different shapes that the shape of cross section of different parts also can allow.Relate to some kinds of electrodes in the structure of this case micro-fluidic chip, its size is the size that is suitable for being placed in the pipeline of relevant position, but, based on as far as possible less with the consideration of aspects such as liquid sample to be measured and reduction reagent loss, described electrode is the minute sized electrode that is suitable for being placed in the kapillary, the shape of each electrode wherein all can be selected arbitrarily shape, and described selected arbitrarily shape is square shape sheet shape, rectangle sheet, strip or round sheet or the like for example.The material of each electrode wherein all can be selected arbitrarily material, and described selected arbitrarily material is gold, silver, platinum or other metal etc. for example.
Relate to several pond shape things in this case microfluidic chip structure, described pond shape thing is pond shape or the scrotiform structure that is used for actual tested blood serum sample, buffer solution and waste liquid storage, its shape of inner chamber of each pond shape thing wherein all can be selected arbitrarily shape, and described cavity shape is cylindrical cavity shape, square column type cavity-like, oblong cavity shape or spherical hollow space shape or the like for example.The size of described pond shape thing can be arbitrarily selected size, still, for can be as far as possible less with liquid sample to be measured and reduce reagent loss, the hypomegetic pond shape thing that described pond shape thing preferably can mate with kapillary.
This case device can further include some annexes certainly, and described annex is multiple tracks electrochemical workstation and microfluidic pump or the like for example, and the art-recognized meanings of the art-recognized meanings of described multiple tracks electrochemical workstation and microfluidic pump is known.Each working electrode that relates in this case microfluidic chip structure and to electrode and contrast electrode etc., can get lines crossed via corresponding special use is respectively connecting with the corresponding interface of described multiple tracks electrochemical workstation.It is to be used for the private cable that each the corresponding interface with each described electrode and described multiple tracks electrochemical workstation is coupled to each other that described special use is being got lines crossed.Described microfluidic pump is exclusively used in and drives micro liquid and flow, described microfluidic pump can with the described pond of selected any one shape thing UNICOM as required.
Can there be several different methods in the specific implementation path of this case micro-fluidic chip, selects the one detailed annotation as follows:
One, superpolymer (polycarbonate: the PC) preparation of the little gold electrode of electroless plating on the cover plate
1, is photomask with quartz/chromium plate, as light source the selective area on PC cover plate surface carried out radiation, generate carboxyl with the ultraviolet light of selecting wavelength.
2, adopt diamines as aminating agent, 1-[3 ,-(N-N-dimethyl amine) propyl group]-the basic carbodiimide hydrochloride of 3-is that coupling agent carries out aminating reaction to the PC cover plate through the selective area irradiation, forms amido link.
3, will immerse in the certain density chlorauric acid solution through the PC cover plate of selective area aminating reaction and react, gold ion is complexed to the amido surface.
4, clean with distilled water after, again it is immersed and reduces 8min in sodium borohydride solution.
5,, with ultrasonic cleaning 30min in the potassium rhodanate solution of its immersion 0.5mol/L, prevented plating again with after the distilled water cleaning.
6, will contain in the plating bath of gold ion, complexing agent and reductive agent formaldehyde through the PC cover plate immersion of cleaning, react 1h down in 45-60 ℃.
7, the cover plate that will be coated with little gold electrode is put into the about 3h of baking oven annealing in process, promptly obtains the many little gold electrode cover plates of superpolymer PC of the present invention.The long 3mm of wherein little gold electrode, wide 3mm, thickness is about 0.1mm.Spacing is about 3mm each other.
Two, the superpolymer (polycarbonate: the PC) making of substrate that has the microchannel
1, makes silicon mold
Draw floor layout, the pattern line width is about 25-100 μ m, adopts the output of high-resolution laser photocomposing system on transparent film, promptly obtains mask.
On substrate surface, deposit one deck silicon nitride film as sacrifice layer by chemical vapour deposition technique, on this substrate, cover one deck light-sensitive emulsion, in baking oven, handle 15-20min for 60 ℃ by the rotation film coating technique; Mask is covered on the substrate, the principle by the exposure imaging with the image transfer on the mask to the optical cement layer of substrate surface; Method by dry etching is processed into the spatial structure with certain depth with the planar figure on the optical cement layer.Can obtain the silicon mold of lobed passage.The silicon mold for preparing is used H successively
2O
2: H
2SO
4Solution (volume ratio is 1: 4), acetone and distilled water clean, to remove its surperficial oxide.
2, adopt pressure sintering to duplicate the microchannel
The PC sheet cut into be about 4cm, the size of wide 3cm.Ultrasonic cleaning is dried naturally, in hot-press arrangement the PC sheet is heated to about softening temperature (110 ℃), applies certain pressure then on silicon mold, and continues about 1min, can suppress microchannel with the silicon mold complementation at the PC sheet.Then, formpiston and the PC substrate that is carved with the microchannel are cooled off the back demoulding together, just obtain required microchannel.
3, on cover plate, make three microminiature pond shape things
Adopting electric drill on different positions, to get out three microminiature pond shape things respectively on the cover plate.
Three, little gold electrode cover plate of superpolymer and the superpolymer substrate that has a microchannel are handled in the cryogenic oxygen plasma atmosphere totally.
Four, will contain Graphene, nm of gold, diagnosing gastric cancer antibody is coated on corresponding little metal working respectively with the mixture solution of macromolecule Nafion solution and makes on the electrode dried overnight.
Five, chip involution
Aim at little gold electrode cover plate of superpolymer and the superpolymer substrate that has the microchannel at microscopically, with this substrate with cover plate is fixed and clamp with two cover glasses, be placed in the baking oven and under uniform temperature, be incubated 15min, can obtain required four-way micro-fluidic chip.
Six, the PC sheet that step 4 is finished is respectively at working electrode, reference electrode and utilize elargol to connect copper sheet as lead to electrode end, treats that elargol seals up hot melt adhesive after dried fully again with fixing copper sheet, promptly finishes the making of whole four-way micro-fluidic chip.
Its relevant each operating parameter of the above-mentioned specific implementation path of this case micro-fluidic chip of detailed annotation can be adjusted according to actual needs.
The using method of this case micro-fluidic chip:
Employing adds Micropump and drives the steady flow in the capillary channel of four-way micro-fluidic chip of liquid stream, utilizes the four-way electrochemical analytical instrument respectively four kinds of diagnosing gastric cancer antigens to be detected.
The concrete detection of this case micro-fluidic chip uses step as follows:
1, adopts the four-way electrochemical analyser, in the microtubule road, feed certain density potassium ferricyanide solution, adding under the Micropump driving, adopt the ampere method to detect the size of current of said process as electron mediator.
2, add actual blood serum sample liquid in the microtubule road, adding under the Micropump driving, various diagnosing gastric cancer antigen molecules are caught by corresponding diagnosing gastric cancer antibody molecule on the working electrode surface in each passage.
3, the diagnosing gastric cancer antigen molecule in diagnosing gastric cancer antibody molecule and the blood serum sample forms immune complex, and certain inhibition is played in the electron transport of the electron mediator potassium ferricyanide, and the electric current that the ampere method detects said process reduces degree.
4, the result who electric current is reduced degree carries out analysis-by-synthesis, obtains the kind and the content of various diagnosing gastric cancer antigen molecules thus, and the cancer of the stomach situation is carried out comprehensive diagnos.
The utility model has the advantages that, at a physical device is integrated four working electrodes that are coated with four kinds of diagnosing gastric cancer antibody materials respectively on the micro-fluidic chip, these four working electrodes are respectively at four kinds of feature diagnostic antigens that detect cancer of the stomach, and the design feature of the integrated structure of this case micro-fluidic chip has determined the use of this chip to help to accelerate diagnosing gastric cancer speed, improves accuracy rate of diagnosis, reduced diagnostic fees usefulness.
[description of drawings]
Fig. 1 is this case micro-fluidic chip embodiment organigram, and what showed is the form of the perspective under the depression angle of this example structure, does not depict described annex among the figure.
Among the figure, 1,2,11 is respectively three pond shape things that installation position is different, the 3rd, the manifold shape fluid passage, 4 represent the installation position difference but the various pipelines and the lateral of formation parallel with one another UNICOM in parallel structure, the 5th, its top layer that is installed in the pipeline is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CEA antibody and macromolecule Nafion solution, the 6th, its top layer that is installed in the pipeline is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA12-5 antibody and macromolecule Nafion solution, the 7th, its top layer that is installed in the pipeline is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA19-9 antibody and macromolecule Nafion solution, the 8th, its top layer that is installed in the pipeline is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA72-4 antibody and macromolecule Nafion solution, the 9th, to electrode, the 10th, contrast electrode.
[embodiment]
In this case embodiment that Fig. 1 showed, this device also is the structure of micro-fluidic chip, comprising that applying is installed in two plate objects together, and two plate objects are respectively the cover plate of micro-fluidic chip and the substrate of micro-fluidic chip.On the position of the mutual applying between two plate objects, be equiped with pipeline, and, three pond shape things.These three pond shape things are respectively the pond shape thing 1 of actual blood serum sample storage, the pond shape thing 2 of buffer solution storage and the pond shape thing 11 of waste liquid storage.One end of described pipeline via manifold shape fluid passage 3 respectively with pond shape thing 1 and pond shape thing 2 UNICOMs, the other end of described pipeline and remaining pond shape thing 11 UNICOMs.This micro-fluidic film-making also is provided with and is installed in the working electrode on the diverse location in the described pipeline in regular turn respectively, to electrode 9 and contrast electrode 10.Working electrode comprises metal electrode and applies the sensitive membrane that the potpourri by Graphene, nm of gold, diagnosing gastric cancer antibody and macromolecule Nafion solution constitutes at metal electrode surface layer.The structure of described pipeline is structure in parallel, and this pipeline that is structure in parallel is made of four lateral parallel connections, and the quantity of working electrode is four, and these four working electrodes are respectively working electrodes 5,6, and 7,8.Wherein, working electrode 5 is that its top layer that is installed in the lateral is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CEA antibody and macromolecule Nafion solution, working electrode 6 is that its top layer that is installed in the lateral is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA12-5 antibody and macromolecule Nafion solution, working electrode 7 is that its top layer that is installed in the lateral is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA19-9 antibody and macromolecule Nafion solution, working electrode 8 are that its top layer that is installed in the lateral is graphene coated, nm of gold, the working electrode of the potpourri of diagnosing gastric cancer CA72-4 antibody and macromolecule Nafion solution.
The annex of the utility model micro-fluidic chip also has annexes (not shown) such as microfluidic pump and multiple tracks electrochemical workstation.Each pond shape thing in this routine structure can be as required with as the microfluidic pump of annex UNICOM by any means.Each working electrode in this routine structure and to electrode and contrast electrode can be respectively via special-purpose separately cable or say get lines crossed respectively with as the corresponding cable interface of the multiple tracks electrochemical workstation of annex or say the interface of getting lines crossed and connect.
Claims (4)
1. the micro-fluidic chip of a diagnosing gastric cancer special use, it is characterized in that: the structure of this micro-fluidic chip comprises that applying is installed in two plate objects together, described two plate objects are respectively a cover plate and a substrate, on the position of the mutual applying between described two plate objects, be equiped with pipeline, three pond shape things and be installed in working electrode in the described pipeline in regular turn respectively, to electrode and contrast electrode, one end of pipeline via the manifold shape fluid passage respectively with wherein two pond shape thing UNICOMs, the other end of pipeline and another remaining pond shape thing UNICOM, described working electrode is made of metal electrode and the sensitive membrane that contains diagnosing gastric cancer antibody that is coated in surface of metal electrode.
2. the micro-fluidic chip of diagnosing gastric cancer special use according to claim 1, it is characterized in that: the structure of described pipeline is structure in parallel, the described pipeline that is structure in parallel is made of four lateral parallel connections, and, the quantity of described working electrode is four, and the installation position of these four working electrodes lays respectively in described four laterals.
3. the micro-fluidic chip of diagnosing gastric cancer special use according to claim 1, it is characterized in that: the diagnosing gastric cancer antibody on these four its top layers of working electrode is respectively that these four kinds of antibody materials are respectively diagnosing gastric cancer antibody: CEA antibody, CA12-5 antibody, CA19-9 antibody and CA72-4 antibody to the antibody of diagnosing gastric cancer antigen energy specificity combination.
4. according to the micro-fluidic chip of any described diagnosing gastric cancer special use among the claim 1-3, it is characterized in that: described pipeline is capillary channel.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011200826263U CN202083692U (en) | 2011-03-16 | 2011-03-16 | Microfluidic chip specially used for gastric cancer diagnosis |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011200826263U CN202083692U (en) | 2011-03-16 | 2011-03-16 | Microfluidic chip specially used for gastric cancer diagnosis |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233939A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | Physical examination examination male's classifiable tumor mark micro flow control chip device |
| CN107233940A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | A variety of everywoman's co-detections of tumor markers in benign micro flow control chip devices |
| CN107233938A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | The tumor markers joint-detection six-channel microfluidic chip device simplified |
-
2011
- 2011-03-16 CN CN2011200826263U patent/CN202083692U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233939A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | Physical examination examination male's classifiable tumor mark micro flow control chip device |
| CN107233940A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | A variety of everywoman's co-detections of tumor markers in benign micro flow control chip devices |
| CN107233938A (en) * | 2016-03-29 | 2017-10-10 | 李榕生 | The tumor markers joint-detection six-channel microfluidic chip device simplified |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111221 Termination date: 20130316 |