CN1462878A - Sensor of chip contaonong microelectrode array - Google Patents
Sensor of chip contaonong microelectrode array Download PDFInfo
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- CN1462878A CN1462878A CN 03129410 CN03129410A CN1462878A CN 1462878 A CN1462878 A CN 1462878A CN 03129410 CN03129410 CN 03129410 CN 03129410 A CN03129410 A CN 03129410A CN 1462878 A CN1462878 A CN 1462878A
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Abstract
A microelectrode array chip sensor on the silicon substrate features that said microelectrode array is composed of interdigital electrodes for smaller gap between positive and negative electrodes and higher transmission speed of electrons. It is prepared by MEMS technique. Its advantages are simple preparing process and high sensitivity.
Description
Technical field
The present invention is a kind of chip sensor of the microelectrode array based on micromachining technology.
Background technology
Chip sensor is the very fast a kind of chemical analysis means of current International Development.It has the advantage of many uniquenesses, and is high as sensitivity, and can be mass-produced (cheap), dynamic response performance is good, size is dwindled greatly, precision height, good reliability, easily be made into the sensor array of multielement, can realize and electronics, the chip that mechanical system integrates etc.Because its range of application is extremely wide, can be used as the number of chemical biology sensor, can be operated in gas, the environment of vacuum and liquid.So it can become the chemical constitution biomolecule is made real-time, online prospecting tools.These characteristics are that other many type sensors are difficult to accomplish.
In past 10 years, Chinese patent about micro-array chip reaches 83, as Chinese patent " the multi-impression fixed-point synthetic method prepares the method for micro-array chip of chemical compound " (application number 98111220, publication number 1193049), it at first overlaps little seal according to the microarray design preparation one of required compound, on each little seal of preparation, be coated with the chemical reactant of last layer correspondence then according to original design, little seal that last order according to design will scribble chemical reactant is stamped on the same substrate one by one successively, utilize the convex-concave of little seal to control the position of chemical reaction on the substrate, on substrate, form required micro-array chip of chemical compound at last." based on the densification compound process for preparing microarray chip of electronic mesh and screen printing technique " (application number 99114435 for another example, notification number 1096627), its characteristics are: (1) has the reaction template of little reaction channel system according to the design of required genetic chip and preparation; (2) utilize the accurate location between little reaction template and the substrate to form the little reaction zone that seals, utilize the method for serigraphy or spray to add corresponding chemical reactant at little reaction zone; (3) according to the order of design,, change template by locating device, the little reaction zone of conversion, thus on substrate, form the required micro-array chip that contains different compound micro units." prepare the method for micro-array chip of chemical compound and by the micro-array chip of chemical compound of this method preparation " (application number 99106790 and for example, publication number 1274758), be to adopt the micro reaction pool motherboard to be positioned at the synthetic method for preparing micro-array chip of chemical compound on the substrate step by step.
In the disclosed micro-array chip invention of this external major part, have many all relevant with biochip, this class chip production method all adopts chemical/biological reaction immobilization of biological substances such as DNA, RNA, cell, tissue etc., and chip itself separates with detection system, the input part that does not relate to.Detect with fluorescence spectrophotometer after for the signal demand that reacts on the chip by fluorescence labeling.As " protein-chip of diagnosing autoimmune diseases " (application number 01126929, publication number 1338633) mainly be to be fixed with being connected on the solid phase carrier by chemical bond-linking with the every index corresponding proteins of autoimmunity disease matter, with described protein example dot matrix in the same chamber of multiple-sample microarray biochip, make multiple-sample microarray biochip the equal dot matrix of each chamber the antigen of above-mentioned many index.Detection method usage flag fluorescence detection, for another example based on the method for making (application number 01127073 of autoantibody repertoire antigen microarray, publication number 1343887), by point sample equipment with the substrate surface of different biomacromolecule antigen points to the above-mentioned modification, make required chip, chip signal is detected, gather sample information with microscope or fluorescent scanning.In addition, application number is 01132492 to 01132499,01135856-01135877, the dozens of patent of 01137660-01137677 all is patents relevant with the paddy gene chip, new expressed sequence tag is used microarray technology and is made expressed sequence tag combination gene chip, and detection means does not all adopt electrochemical measuring technique.
Existing such chip cost height, complicated operation, analyzed object have certain limitation.
Summary of the invention
The purpose of this invention is to provide a kind of novel structure that utilizes prior art for preparing to obtain, simple and practical, easy to operate, highly sensitive, the microelectrode array chip sensor that can be used for multiple detected object.
Microelectrode array chip sensor of the present invention, by on the silicon wafer substrate microelectrode array chip and detection cell form.Microelectrode array on the chip is an interdigitated, and promptly the positive electrode of every pair of electrode, negative electrode are that interdigitated Qian rotation is arranged, and the extension line of microelectrode is positioned at the chip both sides.Detection cell is positioned on the chip, and be one by the encapsulation and the chip of transparent cover plate, the microarray area of the area of detection cell and chip is suitable, be that chip microelectrode array face is the end of detection cell, be covered with transparent cover plate above the detection cell, electrode hole is on transparent cover plate, and the connecting line of contrast electrode and detector is drawn in the hole, import (7) and outlet (8) that the corresponding both sides of detection cell are respectively working fluids.During test sample, testing liquid can be a moving phase, closes if import and export, and testing liquid is non-moving phase.
In the above-mentioned microelectrode array, the distance between the positive and negative microelectrode is better at 10～50 μ m, is not increasing on the basis that technology prepares difficulty, and transducer sensitivity is increased.
Electrode is suitable with 1～20 pair, according to the logarithm that detects the different determining electrode that require.Along with the variation of electrode pair, detection cell size (especially floorage) also makes corresponding changes.
When Chip Packaging, as the encapsulation of detection cell bottom one, the detection cell upper lobes covers completely with transparent cover plate above it on the chip that encapsulation finishes with chip, on every side with the sealing gasket sealing, in case the leakage of testing liquid when flowing.Pass through screw retention between transparent cover plate and the chip.
Adopt microelectron-mechanical process technology (MEMS) during above-mentioned microelectrode array chip sensor production, promptly on silicon chip, use the physical surface film-forming method to make the titanium film of 10～200nm, the golden film of 50～500nm respectively, with the cloudy version of electrod-array system photograph mask, photoetching then, use the silicon dioxide insulating layer of chemical deposition (PECVD) system 100～300nm again, annealing gets final product electrode outlet line with the epoxy resin sealing.The method of determination sensor electric capacity of the present invention is used potential step method, and its principle is as follows: potential step method:
This method is based on electrode system being considered as a simple R C series circuit
After this circuit applied a small voltage pulse (as 50mV), its discharge current was with exponential damping.Expression formula is as follows:
i(t)＝u/Rs?exp(-t/Rs?Cs)
I (t) is the electrode current response, and u is for being applied to electrode surface voltage, and Rs is a bath resistance, and Cs is an electrode capacitance.Response current is taken the logarithm and to time mapping, will be got a linearity curve.Can calculate the electrode capacitance value from this slope of a curve and intercept.The advantage of this method is: can pass through electrode current attenuation measurement electrode capacitance in the time of microsecond level, be convenient to realize hyperchannel, multiple goal clinical detection; Fit procedure is easy simultaneously, is easy to practicability.
The invention provides a kind of microelectrode array chip sensor of silicon substrate surface, this microelectrode array is made up of the interdigitated microelectrode, this structure makes between the positive and negative electrode distance enough little, when having guaranteed between the both positive and negative polarity electro-deposition electropolymerization functional molecular, and the film thickness homogeneous and controllable; Electric transmission speed was far above plane electrode when simultaneously both positive and negative polarity reacted, and measured object is also very fast in the transmission speed of electrode surface, thereby the design of this interdigitated is better than plane electrode, thereby had improved detection sensitivity greatly; This chip utilization MEMS technology makes preparation technology simple, and is easy to operate, and as if the modification of chip surface being made various uses, the present invention can be used for the chip sensor of different purposes and makes.
Table 1-3 has listed sensor experiment result of the present invention.
Wherein table 1 chip array electrode numbering is as follows: left part is divided into A series, from A
1To A
9, A
0, right part is divided into B series, from B
1To B
9, B
0Every pair of electrode specification is as shown in Table 1 on the chip that designs and produces for the first time:
The physical dimension and the area of table 1 interdigited electrode array
The interdigital number electrode area (cm of live width (μ m) spacing (μ m)
2) A12, B12 10 30 4 * 28 * 10
-4A34, B34 10 20 8 * 2 1.6 * 10
-3A56, B56 10 10 8 * 2 1.6 * 10
-3A78, B78 20 20 8 * 2 3.2 * 10
-3A90, B90 20 20 4 * 2 1.6 * 10
-3The interdigital electrode of table 2 different size detects antigen HABP and the sensitivity of LN and the comparison ((nF/cm of responding range
2)/(ng/mL))
Sensitivity ((nF cm
-2
)/(ng mL
-1
))
Responding range (ng mL
-1
)
HABP LN HABP LNA12，B12 0.6106 5.119 0.5-88 0.1-35A34，B34 0.2345 1.989 0.5-188 0.1-55A56，B56 0.2890 1.219 2-88 0.8-35A78，B78 0.4718 2.220 0.5-188 0.1-55A90，B90 0.3287 2.069 0.5-188 0.5-55
Impedance, capacitive reactance and time constant that table 3 interdigited electrode is modified
Impedance capacitive reactance time constant
(KΩ) (nF) (R×C)(μs)A12，B12 85.5 1.5131 129.3A34，B34 52.2 1.9474 101.6A56，B56 120.2 0.8805 105.8A78，B78 38.8 3.6581 142.1A90，B90 49.9 2.6832 133.9
Description of drawings
Fig. 1 is the planimetric map of microelectrode array chip sensor of the present invention.
Fig. 2 is a microelectrode array chip electrode enlarged drawing of the present invention.
Fig. 3 is the detection cell structural drawing that combines with microelectrode array chip sensor of the present invention.
Fig. 4 is the electric capacity response diagram that the present invention detects HABP, LN antigen molecule.
Curve a, b have shown that electrode specification is that live width, distance between centers of tracks are the comb electrode of 20 μ m after having modified HABP, LN antibody molecule respectively, electric capacity response diagram to the HABP in the solution, the detection of LN antigen molecule, in order to provide signal more clearly, this figure ordinate is the capacitance variations value.As can be seen from Figure, the comb electrode that live width and distance between centers of tracks are 20 μ m is 0.5-188ng/ml for the sensing range of HABP molecule, then is 0.1-55ng/ml to the sensing range of LN molecule.The capacitance variations (sensitivity) that it should be noted that the unit area of the same antigen system that the use plane electrode records can not show a candle to table institute column data, and the latter is respectively 2～50ngmL to the responding range of HABP and LN
-1, 1～50ngmL
-1, only be the former 1/4 and 1/10.Be respectively 5ng/ml with the sensitivity of the conventional determining method (being euzymelinked immunosorbent assay (ELISA)) of these two kinds of antigens at present again and compare with 1ng/ml, therefore, the present invention has improved 10 times to the detection sensitivity of antigen.
Among the above-mentioned figure, the 1st, positive electrode, the 2nd, negative electrode, the 3rd, extension line, the 4th, detection cell, the 5th, transparent cover plate, the 6th, electrode hole, the 7th, detection cell import, the 8th, detection cell outlet, the 9th, sealing gasket, the 10th, screw, the 11st, mounting base, the 12nd, live width, the 13rd, electrode separation.
Embodiment
The base material of microchip is a slice n-type or p-type 100 monocrystalline silicon, is of a size of length 3cm, width 2.0cm.The physical surface system of use membrane means vacuum vapour deposition can prepare the layer of metal titanium film at this silicon chip surface, and thickness is 100nm, adopts the gold thin film for preparing a layer thickness 300nm with quadrat method then thereon.The microelectrode array of design as required is made as the cloudy version mask of taking a picture, uses optical etching technology that the metal level (titanium and gold layer) that mask does not hide part is removed.Chemical deposition (PECVD, Plasma Enhance Chemical Vapor Deposition) preparation one layer thickness that uses plasma to strengthen then at the position of removing metal level is the silicon dioxide insulating layer of 150nm.Then, entire chip places oxygen atmosphere to take off fire 30 minutes under high temperature 500 degree.The extension line part of interdigitated microelectrode is sealed with epoxy resin.
With PET film production one length * wide * height is the 50 μ L detection cells (4) of 1cm * 0.5cm * 0.1cm, during Chip Packaging it is together encapsulated, this detection cell is the end with the chip, its upper cover one transparent cover plate (5), and the sealing gasket sealing is arranged, contrast electrode extension line (3) is drawn from cover plate top electrode hole and is connected to detector (11).Fixing with screw (10) between cover plate and the chip, the liquid-inlet of detection cell (7) outlet (8) can be connected with little injector of sensor outside by microchannel.If chip is done following the modification, then can be used for multi-field various sample detection such as biological medicine:
The present invention can modify the surface of the dielectric film of electropolymerization in advance with hepatopathy blood serum designated object monoclonal antibody by bifunctional reagent, also can adopt the method for electrochemical in-situ codeposition that antibody is embedded in the film of electropolymerization, thereby prepare the chip sensor that is used for the different target Detection of antigen.Also can prepare the antibody membrane of poly-adjacent benzene two case dielectric film embeddings and use potential step method to measure the capacitance variations of conjugated antigen.Adopting polymeric sol-gel sets antibody membrane also is one of means of attempting, and perhaps is used for the biology enzyme of stock size electrode surface and the modification of antibody and fixes.
Claims (5)
1, a kind of microelectrode array chip sensor, form by micro-electrode chip on the silicon wafer substrate and detection cell, it is characterized in that the microelectrode array on the chip is an interdigitated, positive electrode (1), the negative electrode (2) that is every pair of electrode is the circuitous arrangement of interdigitated, spacing between the positive and negative electrode is 10～100 μ m, live width is 10-50 μ m, and interdigital several 4-10, the extension line of microelectrode (3) is positioned at the chip both sides; Detection cell (4) is positioned on the chip, and be one by the encapsulation and the chip of transparent cover plate (5), the microarray area of the area of detection cell and chip is suitable, be that chip microelectrode array face is the end of detection cell, above the detection cell transparent cover plate (5), contrast electrode hole (6) is positioned on the transparent cover plate, and import (7), outlet (8) that the corresponding both sides of detection cell are respectively working fluids are by the electroanalyzer detection signal.
2, microelectrode array chip sensor according to claim 1 is characterized in that the distance between the positive and negative microelectrode is 10-50 μ m.
3, microelectrode array chip sensor according to claim 1 is characterized in that microelectrode array is that 1-20 is right on the chip.
4, microelectrode array chip sensor according to claim 1 is characterized in that between detection cell and the transparent cover plate it being sealing gasket (9).
5, microelectrode array chip sensor according to claim 1 is characterized in that between chip and the transparent cover plate it being that screw (10) is fixing.
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|---|---|---|---|
| CNB031294103A CN1195219C (en) | 2003-06-20 | 2003-06-20 | Sensor of chip contaonong microelectrode array |
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|---|---|---|---|
| CNB031294103A CN1195219C (en) | 2003-06-20 | 2003-06-20 | Sensor of chip contaonong microelectrode array |
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| CN1195219C CN1195219C (en) | 2005-03-30 |
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| CN102449467A (en) * | 2009-03-31 | 2012-05-09 | 西门子公司 | Device similar to an electrochemical camera and method for the production and use of the device |
| CN102636530A (en) * | 2012-03-23 | 2012-08-15 | 厦门大学 | Microelectrode array electrolyzer for in-situ cell electrochemical and microscopic detection |
| CN101614729B (en) * | 2008-06-27 | 2013-04-24 | 博奥生物有限公司 | Microelectrode array device and special device for cell manipulation and electrophysiological signal detection |
| CN101426899B (en) * | 2006-04-19 | 2013-08-14 | 戈尔企业控股股份有限公司 | Functional porous substrates for attaching biomolecules |
| CN103675075A (en) * | 2013-11-27 | 2014-03-26 | 复旦大学 | Organophosphorus detection method based on microelectrode chip |
| CN107764880A (en) * | 2017-10-11 | 2018-03-06 | 浙江海洋大学 | A kind of biosensor thin layer electrode of quick detection miocardial infarction protein marker |
| WO2018048362A1 (en) * | 2016-09-09 | 2018-03-15 | King Mongkut's University Of Technology Thonburi (Kmutt) | Process of production of interdigitated array of electrodes and derivatives by screen printing technique |
| WO2018152296A1 (en) * | 2017-02-15 | 2018-08-23 | New Jersey Institute Of Technology | Enhanced sensitivity and specificity for point-of-care (poc) micro biochip |
| CN109831729A (en) * | 2019-01-30 | 2019-05-31 | 武汉大学 | Compact high sensitivity MEMS capacitive sensor |
| CN110006969A (en) * | 2019-04-18 | 2019-07-12 | 哈尔滨理工大学 | A kind of multi-parameter water environment integrated microsensor and preparation method thereof based on electrochemical measuring technique |
| CN111495449A (en) * | 2020-04-01 | 2020-08-07 | 重庆中泽拓邦生物技术有限公司 | Detection chip based on micro-fluidic technology |
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- 2003-06-20 CN CNB031294103A patent/CN1195219C/en not_active Expired - Fee Related
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| CN101426899B (en) * | 2006-04-19 | 2013-08-14 | 戈尔企业控股股份有限公司 | Functional porous substrates for attaching biomolecules |
| CN101614729B (en) * | 2008-06-27 | 2013-04-24 | 博奥生物有限公司 | Microelectrode array device and special device for cell manipulation and electrophysiological signal detection |
| US9995703B2 (en) | 2009-03-31 | 2018-06-12 | Boehringer Ingelheim Vetmedica Gmbh | Device similar to electrochemical camera and method for producing device |
| CN102449467B (en) * | 2009-03-31 | 2014-05-07 | 西门子公司 | Device similar to an electrochemical camera and method for the production and use of the device |
| CN102449467A (en) * | 2009-03-31 | 2012-05-09 | 西门子公司 | Device similar to an electrochemical camera and method for the production and use of the device |
| CN102636530A (en) * | 2012-03-23 | 2012-08-15 | 厦门大学 | Microelectrode array electrolyzer for in-situ cell electrochemical and microscopic detection |
| CN103675075A (en) * | 2013-11-27 | 2014-03-26 | 复旦大学 | Organophosphorus detection method based on microelectrode chip |
| GB2568428A (en) * | 2016-09-09 | 2019-05-15 | King Mongkuts Univ Of Technology Thonburi Kmutt | Process of production of interdigitated array of electrodes and derivatives by screen printing technique |
| WO2018048362A1 (en) * | 2016-09-09 | 2018-03-15 | King Mongkut's University Of Technology Thonburi (Kmutt) | Process of production of interdigitated array of electrodes and derivatives by screen printing technique |
| WO2018152296A1 (en) * | 2017-02-15 | 2018-08-23 | New Jersey Institute Of Technology | Enhanced sensitivity and specificity for point-of-care (poc) micro biochip |
| US11480567B2 (en) | 2017-02-15 | 2022-10-25 | New Jersey Institute Of Technology | Enhanced sensitivity and specificity for point-of-care (POC) micro biochip |
| CN107764880A (en) * | 2017-10-11 | 2018-03-06 | 浙江海洋大学 | A kind of biosensor thin layer electrode of quick detection miocardial infarction protein marker |
| US11020740B2 (en) | 2017-10-24 | 2021-06-01 | New Jersey Institute Of Technology | Microfluidic biochip with enhanced sensitivity |
| CN109831729A (en) * | 2019-01-30 | 2019-05-31 | 武汉大学 | Compact high sensitivity MEMS capacitive sensor |
| CN110006969A (en) * | 2019-04-18 | 2019-07-12 | 哈尔滨理工大学 | A kind of multi-parameter water environment integrated microsensor and preparation method thereof based on electrochemical measuring technique |
| CN111495449A (en) * | 2020-04-01 | 2020-08-07 | 重庆中泽拓邦生物技术有限公司 | Detection chip based on micro-fluidic technology |
| CN111495449B (en) * | 2020-04-01 | 2023-11-03 | 重庆中泽拓邦生物技术有限公司 | Detection chip based on micro-fluidic technology |
| CN111579594A (en) * | 2020-05-27 | 2020-08-25 | 上海交通大学 | Application of room temperature gas sensor in nerve agent detection |
| CN113008959A (en) * | 2021-02-26 | 2021-06-22 | 深圳市西尔曼科技有限公司 | Test electrode, electrode module and detection system |
| CN113008959B (en) * | 2021-02-26 | 2022-07-15 | 深圳市西尔曼科技有限公司 | Test electrode, electrode module and detection system |
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