CN101609063A - A kind of microelectrode array chip sensor that is used for the electro-chemistry immunity detection - Google Patents
A kind of microelectrode array chip sensor that is used for the electro-chemistry immunity detection Download PDFInfo
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Abstract
The invention belongs to sensor technical field, be specially a kind of microelectrode array chip sensor that is used for Electrochemical Detection.The present invention uses silica-base material manufacturing, and chip the is integrated working electrode of Electrochemical Detection, auxiliary electrode and contrast electrode has good signal to noise ratio (S/N ratio).Auxiliary electrode and working electrode constitute ring dish structure, and the distance between auxiliary electrode and working electrode is little, has improved the sensitivity of electrode.A plurality of working electrodes become array distribution, can be used for multi-channel detection.The present invention can be widely used in multi-field detections such as biological medicine.The present invention adopts MEMS technology to make, and technology is simple, and cost is low.
Description
Technical field
The invention belongs to sensor technical field, be specifically related to a kind of microelectrode integrated array chip sensor that is used for the electrochemica biological immune detection.
Background technology
Electrochemical detection method is to utilize electrode that the conversion of signals of the biochemical reaction generation of different material generation is become electric signal, thus the microprocess that reflects between definite material, content of material etc.Difference according to the Electrochemical Detection principle can be divided into current method, potential method, conductance method etc.Since electrochemical method invention, through development constantly, characteristic research to electrode and solution interface has had extraordinary theoretical foundation, and Electrochemical Detection can be applied to analytes such as inorganic ions, organic ion, protein, amino acid, DNA.This also is the reason that the method can be widely used in biology sensor.
Biology sensor is to grow up on the basis of chemical sensor, and it is made up of the sensitive element (molecular recognition elements) and the conversion of signals device that produce signal.The development of biology sensor starts from 1962, and L.c.clark combined electrode and is applied to glucose detection with the film that contains glucose oxidase at that time.Its application is constantly expanded subsequently, and detection method is also updated.According to the difference of detection material, can be divided into several classes such as immunosensor, DNA sensor, enzyme sensor, cell sensor.
Twenty or thirty is in year recently, and the development of MEMS (micro electro mechanical system) (MEMS) process technology further develops biology sensor and biochemical analyzer thereof to the direction of miniaturization, digitizing and high reliability.An urgent demand along with the becoming better and approaching perfection day by day of biotechnology, microelectronics technology rapid development and practical application area, as a kind of hi-tech of multidisciplinary intersection, as a kind of strong analysis tool, biology sensor successfully is applied to fields such as medical science, national defence, environment, food industry and agricultural.
MEMS (micro electro mechanical system) (MEMS) technology is the improvement of electrode to the main influence of electrochemica biological sensor, and the size of electrode has original conventional electrodes to develop into microelectrode.The three-electrode system electrode (contrast electrode, auxiliary electrode, working electrode) that tradition is used respectively can be integrated on the chip; Just can integrated a plurality of electrodes on the sensor component with independent detection function, thus realized that a device can detect multiple biological sample simultaneously.Microelectrode not only has the advantage of miniaturization, the more important thing is that it compares with conventional electrodes and have unrivaled advantage, promptly high steady-state current density, characteristics such as extremely short response time, polarization current is little, mass transfer rate is high, signal to noise ratio (S/N ratio) is big.
Research direction to microelectrode is to make a kind of high stability that has at present, high response characteristic, repeatable high electrode.Be used for the whole integrated chips that are manufactured in of three electrodes of Electrochemical Detection, have very important significance for the stability that detects.The distance that reduces working electrode and auxiliary electrode also becomes a kind of main trend that increases response characteristic.Use Ag/AgCl as contrast electrode, utilize MEMS technology to be integrated among the same device, it is very crucial to improve stability, repeatability.Make a plurality of electrodes simultaneously, and carry out array distribution with a certain structure and many reports and trial are arranged for the raising that detects individual features.Carrying out for the discussion of electrode shape in addition always.
Because integrated electrochemical sensor contrast electrode and auxiliary electrode are introduced, and must consider its shape, the influence of area.In the basic theory of galvanochemistry, make referrals to when working electrode and auxiliary electrode during at the electrolytic cell that is separated from each other, the reaction of auxiliary electrode one side can be ignored substantially.But when working electrode and auxiliary electrode exist together an electrolytic cell, the resultant of reaction of auxiliary electrode one side will have a strong impact on the reaction of working electrode.And point out that in the electrolytic reaction without constant potential, wish that the auxiliary electrode area is more a lot of greatly than working electrode area, the polarization that the outside adds mainly acts on the working electrode.F.Lisdat.D.
Point out also that in " The use ofelectrochemical impedance spectroscopy for biosensing " (on April 16th, 2008) that " Analytical and Bioanalytical Chemistry " magazine is delivered the auxiliary electrode area will be much larger than the area of working electrode.Zhang Zuxun points out that by the method for simulation the current density of little ring electrode is higher than disk electrode in " microelectrode galvanochemistry simulation new method finite analysis method " that magazine " Electroanalytical Chemistry progress " is delivered.
So, on the basis of present MEMS technology, manufacture and design a kind of biochip and will have very large meaning with following characteristics.Working electrode, auxiliary electrode, contrast electrode are integrated in a chip; Increase the area of auxiliary electrode with respect to working electrode; Auxiliary electrode and working electrode structure are loop configuration.
Summary of the invention
It is obvious to the objective of the invention is to propose a kind of response characteristic, and electrochemical properties is stable, the micro-electrode chip sensor that is used for the electro-chemistry immunity detection that biological matching is good.
The micro-electrode chip sensor that is used for the electro-chemistry immunity detection that the present invention proposes is the preparation condition that utilizes existing MEMS technology, has designed a kind of new electrode structure.Three kinds of electrodes that Electrochemical Detection that this chip is integrated is used, i.e. working electrode, auxiliary electrode and contrast electrode.These three kinds of electrodes are made on the same chip by MEMS technology, and its structure as shown in Figure 1.Working electrode is to use the most ripe disk electrode at present, and working electrode is by the Au material with good biocompatibility.Made a plurality of working electrodes in the chip, become array distribution according to certain rule.Auxiliary electrode is the formation that links to each other with the concentric a few end ring arcs of working electrode, the area of auxiliary electrode exerts an influence for the response characteristic of entire chip, the auxiliary electrode that area is bigger for working electrode can make chip operation more stable, but for same chip, the increase of auxiliary electrode area will inevitably make the area of working electrode reduce, the problem that may cause detection signal to die down.Therefore, the auxiliary electrode area should be determined according to the size of working electrode and the sample characteristics of for example of detection.The material of auxiliary electrode can use Au or Pt.Contrast electrode is positioned at the center of entire chip, contrast electrode is a star, its border is made up of multiterminal arc border, and the border of contrast electrode parallel with the border of auxiliary electrode (promptly the distance of two borders on the normal direction of each point equates), the border of contrast electrode and the frontier distance of auxiliary electrode are 100 μ m～200 μ m, and contrast electrode uses Ag/AgCl to complete.
Among the present invention, working electrode becomes array distribution in chip, and the center of circle of all working electrode is on the border of circle that with the chip center is the center of circle; Difference according to test can determine the number of working electrode, working electrode press certain angle evenly distribution on chip, and the number of electrode can be 3～5.The size of working electrode is according to the needs of difference test, different process conditions, and the size of chip etc. determines that working electrode radius of the present invention is selected in 200 μ m～600 mu m ranges.
Auxiliary electrode only is one in a chip, by a plurality of be that the ring arc in the center of circle links to each other and to constitute with the working electrode, and surrounded working electrode.In design, in order effectively to utilize chip area, make adjacent ring arc that lap be arranged, these laps make auxiliary electrode constitute an integral body, the control of the area size of auxiliary electrode mainly realizes by the size that changes ring arc radius, and the ring arc radius that uses 100 μ m～500 μ m has good effect for the steady operation of chip.
Distance between the inboard and corresponding working electrode disk border of auxiliary electrical polar ring arc has significant effects for the sensitivity of Electrochemical Detection, and apart from more little, sensitivity is high more.It is high more that but processing technology requires.The distance that the present invention adopts is 50 μ m～200 μ m.
Three kinds of electrodes on the chip are drawn by electrode cable separately.Chip constitutes sensor by encapsulation with ceramic base, and by the silica gel packaging conductive wire, silica gel constitutes groove along the array chip of chip edge encapsulation and bottom simultaneously, as the detection cell (electrolytic cell) of micro-electrode chip sensor.Integrated chip has been used in this invention, does not need additional electrodes, has guaranteed measuring stability.
Above-mentioned microelectrode array chip sensor production adopts microelectron-mechanical process technology (MEMS), and the present invention realizes by following concrete steps:
1. thermal oxide growth silicon dioxide on P type silicon substrate is as insulation course.
2. use piranha solution (H
2SO
4: H
2O
2=3: 1) clean silica surface, washed with de-ionized water is removed impurity, the oven dry silicon chip; Photoetching uses positive glue to form the working electrode figure.Oxygen plasma sputter bottoming film.
3. sputter PVD Ti is as the adhesion layer of electrode, and sputter Au is as working electrode; Adopt lift-off technology to remove photoresist and form the working electrode figure.
4. photoetching uses positive glue to form the auxiliary electrode figure, and oxygen plasma sputter bottoming film, sputter PVD Ti are as the electrode adhesion layer, and sputter PVD Au or Pt are as auxiliary electrode, and lift-off technology is removed photoresist, forms the auxiliary electrode figure.
5. photoetching uses positive glue to form the contrast electrode figure, and oxygen plasma sputter bottoming film, sputter PVD Ti are as adhesion layer, and sputter PVDAg is as contrast electrode, and lift-off technology is removed photoresist, forms the contrast electrode figure.
6. scribing, routing, silica gel encapsulation.After treating that silica gel solidifies, electrolytic cell adds hydrochloric acid, and contrast electrode adds positive voltage, and electrolytic solution adds negative voltage and constitutes DC channel, forms AgCl by electrochemical reaction on the Ag surface, and contrast electrode completes, and sensor production is finished.
Three kinds of electrodes that galvanochemistry that the galvanochemistry that makes three electrode integrated chips of the present invention are integrated is used are in one, and the interference of having avoided additional electrodes to bring has improved the signal to noise ratio (S/N ratio) of Electrochemical Detection greatly, has strengthened the stability of Electrochemical Detection.Very big simplification has been carried out in use operation for device, has improved the portability of device, helps the commercialization of device.
Working electrode of the present invention can carry out multi-channel detection according to the needs that detect, and can detect simultaneously several samples, has greatly improved the efficient that detects.
The annular auxiliary electrode that the present invention makes is compared traditional circular electrode and is had higher response current, and the area of auxiliary electrode increases, and has very important help for the stability that improves device.The design of ring electrode also makes the area that effectively utilizes of chip obtain enhancing.
The present invention reduces the distance of auxiliary electrode and working electrode in process conditions, has improved the precision of Electrochemical Detection.
The present invention uses ceramic base to encapsulate, and uses insulation silica gel to encapsulate, and has guaranteed the insulating property of encapsulation on the one hand, and silica gel and galvanochemistry three electrode integrated chips constitute electrolytic cell on the other hand, for testing has brought very big convenience.
Description of drawings
Fig. 1 is the electrode shape figure
Fig. 2 is the sensor component structure
Number in the figure: the 1st, working electrode, the 2nd, auxiliary electrode, the 3rd, contrast electrode, the 4th, ceramic base, the 5th, encapsulation silica gel, 6 is chip.
Embodiment
The invention will be further described below in conjunction with drawings and Examples.Embodiment provided detailed embodiment and specific operation process, but protection scope of the present invention is not limited to embodiment being under the technology prerequisite with the present invention program.
1. the silicon dioxide of thermal oxide growth 600nm on P type silicon substrate is as insulation course.
2. use piranha solution (H
2SO
4: H
2O
2=3: 1) clean silica surface, washed with de-ionized water is removed impurity, the oven dry silicon chip; Photoetching uses positive glue to form the working electrode figure, and the working electrode radius is 500 μ m.Oxygen plasma sputter bottoming film.
3. the Ti of sputter PVD 20nm, the Au of 300nm; Adopt lift-off technology to remove photoresist and form the working electrode figure, the working electrode radius is 500 μ m.
4. photoetching uses positive glue to form the auxiliary electrode figure, oxygen plasma sputter bottoming film, sputter PVD 20nm Ti, 300nm Pt, lift-off technology is removed photoresist, form auxiliary electrode figure, the ring of shape auxiliary electrode directly is 200 μ m, and the distance of auxiliary electrode and working electrode is 100 μ m.
5. photoetching uses positive glue to form the contrast electrode figure, oxygen plasma sputter bottoming film, sputter PVD 20nm Ti, 400nmAg, lift-off technology is removed photoresist, form the contrast electrode figure, the distance on the border of contrast electrode and auxiliary electrode border is 100 μ m.
6. scribing, routing, silica gel encapsulation.After treating that silica gel solidifies, electrolytic cell adds hydrochloric acid, and the ginseng electrode adds positive voltage 0.5V, and electrolytic solution adds negative voltage and constitutes DC channel, and energising 5s forms AgCl by electrochemical reaction on the Ag surface, and the ginseng electrode completes, and sensor production is finished.
After sensor production is finished, use electrochemical analyser, adopt cyclic voltammetry to test, the chip that encapsulation is finished carries out the test of bare electrode.Electrochemical analyser auxiliary electrode and working electrode line link to each other with working electrode with the auxiliary electrode of sensor chip respectively and constitute path, the contrast electrode lead be connected the chip contrast electrode reference voltage be provided.The mixed solution that in the electrolytic cell of sensor, adds 10mM PBS solution and 10mM ferrocene methanol, PBS is for supporting dielectric, ferrocene methanol is a probe.With 0.1V/s, scan at-0.5V～0.5V, record cyclic voltammetry curve.Sensor of the present invention is tested, and the peak value that obtains oxidation peak is 3.43 μ A, and the peak value of reduction peak is 3.34 μ A, can obtain its peak current density by calculating and be respectively 0.437mA/cm
2And 0.425mA/cm
2Illustrate that this sensor has higher sensitivity.
The present invention can be used for the detection of multi-field various samples such as biological medicine.The present invention can by the electrochemical in-situ codeposition method antibody is embedded in the film of electropolymerization, thereby can be used for the chip sensor that different target detects.The antibody moiety dielectric film that also can prepare the embedding of poly-o-phenylenediamine dielectric film carries out electrochemical analysis.Perhaps adopt the method for self assembly molecule that antibody is fixed to working electrode, thereby constitute biology sensor with different testing goals.
Claims (3)
1. one kind is used for the microelectrode array chip sensor that electro-chemistry immunity detects, the chip of this sensor by the MEMS process technology integrated three electrodes that use of Electrochemical Detection: working electrode, auxiliary electrode, contrast electrode, it is characterized in that described working electrode has 3～5, each working electrode is a disc, disc radius is 200 μ m-600 μ m, it is on the circle in the center of circle that the center of circle of working electrode is positioned at the chip center, evenly distributes; Described auxiliary electrode constitutes by linking to each other with concentric several sections rings of working electrode arcs, has surrounded working electrode, and as a whole, and the radius that each section encircles arc is 100 μ m～500 μ m; Described contrast electrode is positioned at the center of chip, and the border of contrast electrode is made up of the multistage arc, and the border of contrast electrode is parallel with the border of auxiliary electrode, and the distance between two boundary electrodes is 100 μ m～200 μ m; Three kinds of electrodes are drawn by lead; Chip Packaging is on ceramic base, and by silica gel packaging insulating lead, silica gel encapsulates along chip edge simultaneously, forms groove and constitutes electrolytic cell, and microelectrode array chip is the end of electrolytic cell.
2. microelectrode array chip sensor according to claim 1 is characterized in that each distance of encircling between the inboard and corresponding working electrode disk border of arc of auxiliary electrode is 50 μ m-200 μ m.
3. microelectrode array chip sensor according to claim 1 is characterized in that described worker organizes electrode and adopts Au, and described auxiliary electrode adopts Au or Pt, and affiliated contrast electrode adopts Ag/AgCl.
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| CN200910054886.7A CN101609063B (en) | 2009-07-16 | 2009-07-16 | Microelectrode array chip sensor for electrochemical immunological detection |
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| CN113189168A (en) * | 2021-03-31 | 2021-07-30 | 广州钰芯传感科技有限公司 | Microelectrode integrated sensing chip and application thereof in rapid detection of organic pollutants in water |
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