CN103627631A - Polypyrrole/graphene decorated dual-mode nerve microelectrode array chip and preparation method thereof - Google Patents
Polypyrrole/graphene decorated dual-mode nerve microelectrode array chip and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a polypyrrole/graphene decorated dual-mode nerve microelectrode array chip and a preparation method thereof. The chip comprises an insulative substrate, multiple pads, multiple strip leads, four groups of microelectrode arrays; the multiple pads are distributed at the periphery of the insulative substrate at equal intervals; the four groups of the microelectrode arrays are symmetrically distributed at the center position of the insulative substrate surface; the electrodes in the microelectrode arrays respectively extends to the peripheral edge of the insulative substrate by the stripe leads connected with the pads via ends; and the surface of the work electrode in the microelectrode array is decorated with a composite sensitive film material. The invention also provides a method for preparing the chip. The chip provided by the invention helps to overcome the disadvantages that conventional nerve microelectrodes are high in impedance and an in-vitro biological tissue is not easy to contact with the electrode, has the advantages of being low in impedance and good in biological compatibility, and the existing of graphite increases the roughness and the nonuniformity of the electrode surface. The function integration of the chip provided by the invention is beneficial to granting and detection of in-vitro brain slice cells and cultured nerve cells.
Description
Technical field
The present invention relates to micro-processing technology and the nano-modified technical field of biosensor, neural microelectrode array chip and the preparation method of detecting of bimodulus that especially a kind of polypyrrole/graphene composite material is modified.
Background technology
Neural system is the tract consisting of nervous tissue, is also one of of paramount importance system of complexity the most in human body, and it has comprised billions of neurones and multiple sensory information pass through mechanism.Single neuronic information transmission is completed jointly by electrical signal and mediator signal, and this two classes signal plays an important role in neural system.Therefore study the synchronous detection of Electrophysiology signal and mediator electrochemical signals, for neural further research, have great significance.
Graphene is world the hardest the thinnest nano material at present, and under normal temperature, its electronic mobility surpasses ratio nano carbon pipe or silicon crystal, and resistivity is only about 10
-6Ω cm, is the current material of resistivity minimum in the world, and based on above characteristic, Graphene has fabulous application prospect.Pyrrole monomer can more promptly be oxidized to polypyrrole under the existence of oxygenant, polypyrrole is compared with other polymers has advantages of high, the easy film forming of specific conductivity, softness, good biocompatibility, and pyrroles is easy to other materials compound, so polypyrrole is being employed aspect electrode modification, ion detection widely.
Traditionally, patch clamp, glass microelectrode etc. is the common technology that people detect neurocyte electricity physiological signal, but uses these technology positioning of electrode difficulties, complex operation, and easily cause cell injury, making cannot long term monitoring.In recent years, development along with MEMS (micro electro mechanical system) (MEMS) processing technology, the invention of microelectrode array chip makes to the long-term non-destructive monitoring of neurocyte, to provide possibility at body or in vitro, as the MEA chip of German Multichannel company exploitation, the Michigan electrode of University of Michigan's research and development etc.These electrodes can be realized the synchronous detection of colony's neurocyte bioelectrical activity, yet the microelectrode array before using does not have integrated electrochemical to detect the function of neurotransmitter and the signal to noise ratio of picked up signal is low, this is because the material sensitivity of bare electrode or modification is not high, detectability is low, is difficult for contacting with in vitro tissue.
Summary of the invention
The advantage of Graphene and the deficiencies in the prior art, the neural microelectrode array chip and preparation method thereof that detects of bimodulus that provides a kind of polypyrrole graphene composite material to modify are provided for above-mentioned polypyrrole.
For realizing this purpose, according to an aspect of the present invention, propose the neural microelectrode array chip that detects of bimodulus that a kind of polypyrrole/graphene composite material is modified, this chip comprises: dielectric base 1, a plurality of pad 2, a plurality of leads 3, four groups of microelectrode arrays 4, wherein:
The carrier that described dielectric base 1 is whole chip;
Described a plurality of pad 2 is distributed in the periphery of described dielectric base 1 equally spacedly;
The center position on described dielectric base 1 surface is distributed with four groups of microelectrode arrays 4 symmetrically, is distributed with four symmetric matrix formulas circular microelectrode that distribute, that made by conductive film material in every group of microelectrode array;
Four groups of microelectrode arrays 4 comprise a reference electrode 5, and remaining is working electrode, and any one in described working electrode is to electrode;
Described reference electrode 5 is positioned at the center of whole dielectric base 1, at least large order of magnitude of other working electrodes of its Area Ratio;
All electrodes in microelectrode array 4 are respectively by 3 edges that extend to described dielectric base 1 that go between, and the end of described lead-in wire 3 is connected with corresponding pad 2, and the quantity of the quantity of described lead-in wire 3, the quantity of all electrodes and described pad 2 is all identical;
The surface coverage of described lead-in wire 3 has insulation layer;
The finishing of the working electrode of described microelectrode array 4 has composite sensing mould material 7.
According to a further aspect in the invention, also propose the neural preparation method who detects microelectrode array chip of bimodulus that a kind of polypyrrole/graphene composite material is modified, said method comprising the steps of:
Step 1, the surface of using acetone, ethanol or washed with de-ionized water dielectric base;
Step 4, the Ti Seed Layer that is 30-50nm in surface sputtering a layer thickness of described photoetching agent pattern, to increase the adhesivity of Pt conductive membrane layer and described dielectric base;
Step 8, by the method for photoetching and plasma etching, exposes microelectrode array and pad, but retains the insulation layer that all wire surfaces cover;
Step 9, is used polypyrrole graphene composite material to modify the chip that described step 8 obtains.
The neural microelectrode array chip that detects of bimodulus that polypyrrole graphene composite material provided by the invention is modified, the function of multichannel nerve electric physiological detection, Electrochemical Detection is integrated in one, and on nerve microelectrode array, has modified PPy/GR composite sensing material.During detecting, the neurocyte that present method can detect, cultivate in vitro brain sheet obtains signal to noise ratio high, highly sensitive nerve signal, this is for the inherent mechanism of nerve information coding, transmission, and pathogenetic further investigation of some neuropsychiatric diseases, significant.
Accompanying drawing explanation
Fig. 1 is the neural microelectrode array chip structural representation that detects of bimodulus that polypyrrole graphene composite material is modified according to an embodiment of the invention;
Fig. 2 is the local enlarged diagram of microelectrode array according to an embodiment of the invention;
Fig. 3 is the neural process flow sheet that detects microelectrode array chip preparation method of bimodulus that polypyrrole graphene composite material is modified according to an embodiment of the invention.
Embodiment
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in more detail.
Fig. 1 is the neural microelectrode array chip structural representation that detects of bimodulus that polypyrrole/graphene composite material is modified according to an embodiment of the invention, as shown in Figure 1, bimodulus that described polypyrrole graphene composite material is modified is neural to be detected microelectrode array chip and comprises dielectric base 1, a plurality of pad 2, a plurality of leads 3, four groups of microelectrode arrays 4, wherein:
The carrier that described dielectric base 1 is whole chip, the material selection hard transparent of described dielectric base 1, the insulating material of good biocompatibility, such as silica glass, polyvinyl chloride or polycarbonate, described dielectric base 1 be shaped as square, the length of side is 50mm～80mm, thickness is 1mm～2mm, and certainly, described dielectric base 1 also can be designed to rectangle; In an embodiment of the present invention, described dielectric base 1 is substrate of glass, and the length of side is 50mm, and thickness is 1.5mm.
Described a plurality of pad 2 is distributed in the periphery of described dielectric base 1 equally spacedly, and in an embodiment of the present invention, described pad 2 is square for the length of side is 0.25mm;
Fig. 2 is the local enlarged diagram of chip microelectrode array.As shown in Figure 2, the center position on described dielectric base 1 surface is distributed with four groups of microelectrode arrays 4 symmetrically, in every group of microelectrode array, be distributed with four symmetric matrix formulas circular microelectrode that distribute, that made by conductive film material, the diameter of described microelectrode is 20 μ m～40 μ m, be preferably 20 μ m, interelectrode distance is 200 μ m, and described interelectrode distance is the distance between the center of electrode between two.
Four groups of microelectrode arrays 4 comprise a reference electrode 5, and remaining is working electrode, and further, any one in described working electrode is to electrode, such as specifying the electrode of Reference numeral 6 indications in Fig. 1, are to electrode; Described reference electrode 5 is positioned at the center of whole dielectric base 1, at least large order of magnitude of other working electrodes of its Area Ratio, and preferably, the diameter of described reference electrode 5 is 100 μ m or 150 μ m.
All electrodes in microelectrode array 4, comprise reference electrode, to electrode and other working electrodes, respectively by 3 edges that extend to described dielectric base 1 that go between, and the end of described lead-in wire 3 is connected with corresponding pad 2, to facilitate with external circuit, is connected.The quantity of the quantity of described lead-in wire 3, the quantity of all electrodes and described pad 2 is all identical.
Electrode in described microelectrode array 4, go between 3, the material of pad 2 is conductive film material, described conductive film material is metal or the metallic compound of good biocompatibility, such as macromolecule conducting material or titanium platinum film, its thickness is greater than 300nm, to guarantee that its physical strength can bear the pressure that standard electronic components and parts Elastic metal probe is caused; The surface of described lead-in wire 3 is also coated with insulation layer, and the organic or inorganic insulating material that the material that described insulation layer is used is good biocompatibility, such as silicon-dioxide, silicon nitride, nitrogen-oxygen-silicon, SU8, polyimide or polyphenylene ethyl.
The surface of the working electrode of described microelectrode array 4 is modified with composite sensing mould material 7 by plating or MEMS technique, described composite sensing mould material 7 can be selected from gold, platinum, titanium nitride, nanometer platinum black, indium tin oxide or titanium platinum film, in an embodiment of the present invention, the matrix material that described composite sensing mould material 7 is polypyrrole PPy/ Graphene GR.
In use, by the neurocyte of the nervous tissue of in vitro animal or cultivation, with described microelectrode array 4 close contacts, then in conjunction with supporting detection system, the bimodulus that can carry out animal exsomatizednerve information or culturing cell detects and correlative study described chip.
According to a further aspect in the invention, the neural preparation method who detects microelectrode array chip of bimodulus who also provides a kind of polypyrrole/graphene composite material to modify, as shown in Figure 3, said method comprising the steps of:
Step 1, the surface (as shown in Fig. 3 (a)) of using acetone, ethanol or washed with de-ionized water dielectric base;
Step 4, the Ti Seed Layer that is 30-50nm in surface sputtering a layer thickness of described photoetching agent pattern, to increase the adhesivity (as shown in Fig. 3 (d)) of Pt conductive membrane layer and described dielectric base;
Step 8, by photoetching and plasma etching (such as SF
6plasma etching) method, exposes microelectrode array and pad, but retains the insulation layer (as shown in Fig. 3 (h)) that all wire surfaces cover;
Step 9, is used polypyrrole graphene composite material to modify the chip that described step 8 obtains.
Described step 9 is further comprising the steps:
Step 91, adopts improved Hummer method to prepare graphene oxide;
Step 92, preparation contains pyrroles 0.05M (mol/l, mole every liter), PSS (sodium polystyrene sulfonate) 0.05M, the electrolytic solution of graphene oxide 1.0g/l;
Step 93, is used the electrolytic solution that described step 92 is prepared on microelectrode array, to carry out electropolymerization, and such as adopting timing voltage method, electric quantity density is controlled at 0.5-4.0Ccm
-2, the electric quantity density of every square centimeter is controlled at 0.5-4.0 coulomb, thereby has realized the PPy/GR matrix material modification of microelectrode array;
Step 94, after described electropolymerization has been modified, rinses it with a large amount of deionized waters at once, and the oven dry of spending the night in the vacuum drying oven of 70 degrees Celsius, then the chip making is stored in moisture eliminator.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; institute is understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any modification of making, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. the bimodulus nerve of a polypyrrole/graphene composite material modification detects microelectrode array chip, it is characterized in that, this chip comprises: dielectric base (1), a plurality of pad (2), a plurality of leads (3), four groups of microelectrode arrays (4), wherein:
The carrier that described dielectric base (1) is whole chip;
Described a plurality of pad (2) is distributed in the periphery of described dielectric base (1) equally spacedly;
The center position on described dielectric base (1) surface is distributed with four groups of microelectrode arrays (4) symmetrically, is distributed with four symmetric matrix formulas circular microelectrode that distribute, that made by conductive film material in every group of microelectrode array;
Four groups of microelectrode arrays (4) comprise a reference electrode (5), and remaining is working electrode, and any one in described working electrode is to electrode;
Described reference electrode (5) is positioned at the center of whole dielectric base (1), at least large order of magnitude of other working electrodes of its Area Ratio;
All electrodes in microelectrode array (4) extend to the edge of described dielectric base (1) respectively by lead-in wire (3), and the end of described lead-in wire (3) is connected with corresponding pad (2), and the quantity of the quantity of described lead-in wire (3), the quantity of all electrodes and described pad (2) is all identical;
The surface coverage of described lead-in wire (3) has insulation layer;
The finishing of the working electrode of described microelectrode array (4) has composite sensing mould material (7).
2. chip according to claim 1, is characterized in that, the material of described dielectric base (1) is the insulating material of hard transparent, good biocompatibility.
3. chip according to claim 1, is characterized in that, described dielectric base (1) be shaped as square or rectangle, thickness is 1mm～2mm, when described dielectric base (1) be shaped as square time, the length of side is 50mm～80mm.
4. chip according to claim 1, is characterized in that, described pad (2) is square for the length of side is 0.25mm.
5. chip according to claim 1, is characterized in that, the diameter of described microelectrode is 20 μ m～40 μ m, and interelectrode distance is 200 μ m; The diameter of described reference electrode (5) is 100 μ m or 150 μ m.
6. chip according to claim 1, it is characterized in that, the material of the electrode in described microelectrode array (4), lead-in wire (3), pad (2) is conductive film material, the metal that described conductive film material is good biocompatibility or metallic compound.
7. chip according to claim 1, is characterized in that, the organic or inorganic insulating material that the material that described insulation layer is used is good biocompatibility.
8. chip according to claim 1, is characterized in that, modification mode is for electroplating or MEMS technique; Described composite sensing mould material (7) is gold, platinum, titanium nitride, nanometer platinum black, indium tin oxide or titanium platinum film.
9. the neural preparation method who detects microelectrode array chip of bimodulus that polypyrrole/graphene composite material is modified, is characterized in that, said method comprising the steps of:
Step 1, the surface of using acetone, ethanol or washed with de-ionized water dielectric base;
Step 2, spin coating one deck positive photoresist in the dielectric base of cleaning in surface process, thickness is greater than three times of plan sputter conductive membrane layer;
Step 3, forms the photoetching agent pattern of pad, lead-in wire, microelectrode array after photoetching development;
Step 4, the Ti Seed Layer that is 30-50nm in surface sputtering a layer thickness of described photoetching agent pattern, to increase the adhesivity of Pt conductive membrane layer and described dielectric base;
Step 5, the microelectrode conductive membrane layer that is 250nm-350nm in described Ti Seed Layer surface sputtering a layer thickness.
Step 6, adopts stripping technology to remove unnecessary Ti Seed Layer and conductive membrane layer, leaves required microelectrode array, lead-in wire and pad;
Step 7, by plasma enhanced chemical vapor deposition silicon-dioxide, silicon nitride or nitrogen-oxygen-silicon, or the method for spin coating SU8, polyimide or polyphenylene ethyl, at the substrate surface for preparing conductive membrane layer, cover insulation layer;
Step 8, by the method for photoetching and plasma etching, exposes microelectrode array and pad, but retains the insulation layer that all wire surfaces cover;
Step 9, is used polypyrrole graphene composite material to modify the chip that described step 8 obtains.
10. chip according to claim 9, is characterized in that, described step 9 is further comprising the steps:
Step 91, prepares graphene oxide;
Step 92, preparation contains pyrroles 0.05M, sodium polystyrene sulfonate 0.05M, the electrolytic solution of graphene oxide 1.0g/l;
Step 93, is used the electrolytic solution that described step 92 is prepared on microelectrode array, to carry out electropolymerization;
Step 94, after described electropolymerization has been modified, rinses it with a large amount of deionized waters at once, and the oven dry of spending the night in the vacuum drying oven of 70 degrees Celsius, then the chip making is stored in moisture eliminator.
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| CN105460882A (en) * | 2015-12-04 | 2016-04-06 | 中国科学院上海微系统与信息技术研究所 | Graphene three-dimensional microelectrode array chip, and method and application thereof |
| CN105628757A (en) * | 2015-12-30 | 2016-06-01 | 中国科学院电子学研究所 | ORP sensing chip based on MEMS and manufacturing method of ORP sensing chip |
| CN108254414A (en) * | 2018-03-09 | 2018-07-06 | 国家纳米科学中心 | A kind of flexible in vitro micro- raceway groove microelectrode array integrated chip and its preparation method and application |
| CN110632160A (en) * | 2019-09-23 | 2019-12-31 | 南京市食品药品监督检验院 | Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection |
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| CN108254414B (en) * | 2018-03-09 | 2024-02-02 | 国家纳米科学中心 | Flexible in-vitro micro-channel microelectrode array integrated chip and preparation method and application thereof |
| CN108254414A (en) * | 2018-03-09 | 2018-07-06 | 国家纳米科学中心 | A kind of flexible in vitro micro- raceway groove microelectrode array integrated chip and its preparation method and application |
| CN110632160A (en) * | 2019-09-23 | 2019-12-31 | 南京市食品药品监督检验院 | Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection |
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| CN111348616A (en) * | 2020-03-06 | 2020-06-30 | 清华大学 | Implantable neural electrode and preparation method thereof |
| CN112716497A (en) * | 2020-12-21 | 2021-04-30 | 中国科学院空天信息创新研究院 | Micro-nano electrode array chip for positioning single-cell horizontal brain function and preparation method thereof |
| CN112716497B (en) * | 2020-12-21 | 2023-07-04 | 中国科学院空天信息创新研究院 | Micro-nano electrode array chip for single-cell horizontal brain function positioning and preparation method thereof |
| WO2023187217A1 (en) * | 2022-04-01 | 2023-10-05 | Fepod Oy Ltd | Optically transparent microelectrode arrays for electrochemical and electrophysiological measurements or stimulation |
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Application publication date: 20140312 |