CN104760922B - A kind of ultra micro planar electrode array sensor and preparation method thereof - Google Patents
A kind of ultra micro planar electrode array sensor and preparation method thereof Download PDFInfo
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Abstract
The invention discloses the preparation method of a kind of bimodulus (electricity physiological signal and electrochemical signals both of which signal) ultra micro planar array sensor for quantitative determining nerve cell Quantum neural computing.Prepare ultra micro plane electrode (0.5～5 μm) in conjunction with two-layer wiring design, stepping photoetching process in high precision and subregion isolation design, be oriented nano-modified, bio-compatible sex modification on electrod-array surface and method that specific recognition enzyme modification combines prepares bimodulus plane ultramicroelectrode array sensor.The method using micro electro mechanical system (MEMS) technology, nano-modified technology and bio-modification technology to combine prepares bimodulus plane ultramicroelectrode array.Overcome electrode site size (10～50 μm) bigger than normal of conventional plane microelectrode and the limitation of the Single locus detection of bar-shaped carbon fiber electrode.The method prepares ultra micro planar electrode array and has that electrode site is little, measuring point is many, to nerve cell not damaged, neurotransmitter Quantum neural computing and the bimodulus information of electro physiology action potential signal that can detect multiple nerve cells the most in real time.
Description
Technical field
The present invention relates to biology sensor, Electroanalytical Chemistry micro-nano preparing technical field, be a kind of permissible
The multiple electrochemical nerve mediator signal of detection cellular electrophysiologicalsensor and the many synaptic sites of cell is super simultaneously
Micro-array sensors and preparation method thereof.
Background technology
The neurotransmitter that nerve cell axons tip vesica stores is quite stable, in units of vesica
Empty one's purse to discharge, be referred to as the release of quantum formula.The arrival of one-off current potential, can make about 200～300
The content discharge of individual vesica.It is to understand god that nerve cell quantization neurotransmitter regulator carries out real time record
Through signal conduction, the important direct mode of mechanism of action.Monitoring cell Quantum neural computing must possess superelevation
The analytical technology of resolution, ultra-small volume is asked in sensitivity, high selectivity, high time resolution, high-altitude.Pass
The method of system detection nerve cell Quantum neural computing is essentially patch-clamp and coordinates carbon fiber electrode technology, generally
It is only capable of obtaining the data of a small amount of 12 passages, and electrode location difficulty, complex operation, it is impossible to long
Phase monitors, and largely limits the detection to cell Quantum neural computing.
Along with the development of MEMS (MEMS) process technology, microelectrode array
(microelectrode, MEA) provides the method for a kind of site long-term, many monitoring cell, conventional
MEA electrode diameter in 10～50 μm, much larger than the Quantum neural computing size of vesica and cynapse, although
Individual cells can be navigated to, but for being positioned to detect the Quantum neural computing of synaptic vesicle, in space
The most not enough with in temporal resolution, it is impossible to distinguish the space of cell different loci release on single electrode
Difference, it is difficult to measure real-time Quantum neural computing signal.It is thus desirable to develop smaller electrode, with
Realize the monitoring of Single cell release time-space resolution and be deep into synapse cell gap and study, deeper
Cell releasing mechanism is inquired on level.
Summary of the invention
The present invention is directed to the real-time detection of cell and synaptic vesicle Quantum neural computing micro substance, solve at present
Problem: can only detect individual cells or vesica, once can only detect a kind of neurotransmitter, electrode size
Excessive, it is difficult to detect in cynapse size；Design preparation is a kind of, and site is little, measuring point is many, right
Nerve cell not damaged, multiple measuring point can be detected simultaneously (detect multiple simultaneously on two dimension yardstick
Cell, various neurotransmitters) ultra micro planar electrode array sensor.
According to an aspect of the present invention, it provides a kind of ultra micro planar electrode array sensor, its feature
It is: include ultra micro planar electrode array and composite function film layer, wherein said ultra micro plane electrode battle array
Arrange by dielectric base, microelectrode array, electrode, reference electrode, contact conductor and contact constituted,
Described microelectrode array includes multiple, and on a dielectric base, multiple microelectrode arrays can share ginseng in distribution
Than electrode with to electrode；Described composite function film layer include nano-modified, bio-compatible modify, biological
Special layer modifies the decorative layer combined, and its subregion is modified at ultra micro planar electrode array.
According to a further aspect of the invention, it provides the preparation of a kind of ultra micro planar electrode array sensor
Method, it is characterised in that comprise the steps:
Step 1, at spin coating one layer photoetching glue in the dielectric base of surface clean, after photoetching development
Form lead-in wire and microelectrode array pattern；
Step 2, microelectrode array patterned surfaces sputter one layer of microelectrode conductive membrane layer；
Step 3, use stripping technology remove unnecessary microelectrode conductive membrane layer, stay required electrode,
Lead-in wire and contact, basis of formation metal electrode array；
Step 4, on underlying metal electrod-array surface by plasma enhanced chemical vapor deposition exhausted
Edge layer；
Step 5, there is the first area of basic electrode array of insulating barrier in deposition, use two-layer wiring
Ground floor domain carry out secondary photoetching, use stepping photoetching process to form ultramicroelectrode array Centered Graphs
Shape, the method then using plasma etching, expose the first ultramicroelectrode array and contact, retain
The insulating barrier that all wire surfaces cover；
Step 6, there is the second area of basic electrode array of insulating barrier in deposition, repeat step 5 quarter
Erosion exposes the second ultramicroelectrode array and corresponding contacts；
Step 7, employing SU-8, PDMS or polyimides preparation have on the chip of ultramicroelectrode array
By being commonly lithographically formed dam type partitioned organization.
The ultra micro planar array sensor of the present invention relatively has with existing bar-shaped ultramicroelectrode sensor ratio
Following advantage:
Cell detects after growing attachment on micro-planar array sensor, attaches more preferably, the most in situ,
Non-invasive detection.Without location of wasting time and energy.
Ultra micro planar array sensor can detect multiple cell simultaneously, multiple synaptic sites vesica is released
Put.
One sensor of ultra micro planar array sensor can detect various neurotransmitters.
Cell uses micro-nano technique to prepare at micro-planar array sensor, can prepare in batches, and performance is more equal
One, stable.
This ultra micro planar array sensor compares with existing Array microelectrode and has the advantage that
1, ultra micro planar array sensor electrode site is less, size range 0.5 μm-5 μm, more connects
Being bordering on the size range of synapse cell vesica, diffusion is rapid, the beneficially Transient detection of vesica Quantum neural computing.
2, ultra micro planar array sensor uses isolated area designing technique and specific modification technique, can
Realize various neurotransmitters detection.
The ultra micro planar array sensor of the present invention uses the preparation of micro electronmechanical process technology to combine nanometer and repaiies
Decorations technology, can realize the lossless real-time detection to cell, can be used for the unicellular and slender of multiple site
The original position of the various neurotransmitters of born of the same parents' many synaptic vesicles Quantum neural computing detects in real time, and can realize right
Same cell detects while carrying out different stimulated, contrasts detection and statistical analysis simultaneously.
This high density ultra micro planar electrode array of the present invention has area little (0.5 μm～2 μm electricity
Site, pole diameter), measuring point many, to nerve cell not damaged, can two dimension yardstick on examine simultaneously
Survey the neurotransmitter detection of the nerve cell vesica of the most up to a hundred measuring points.Height due to electrod-array
Close characteristic, may further determine that the spatial positional information that single nerve cell is secreted.Carry out partitioned organization to set
Meter, can detect various neurotransmitters on same sensor array simultaneously.
Above-mentioned various purposes, method, feature and the advantage of the present invention, by below in conjunction with the accompanying drawings and real
Execute example and can obtain more detailed explanation.
Accompanying drawing explanation
Fig. 1 is the preparation technology flow chart of ultra micro planar electrode array sensor in the present invention；
Fig. 2 is the double-deck fabric swatch Local map of ultra micro planar electrode array sensor in the present invention；
Fig. 3 is the individual layer fabric swatch partial plan layout of ultramicroelectrode array sensor in the present invention；
Fig. 4 is the ultra micro plane electrode micrograph that in the embodiment of the present invention, platinum black is modified.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with concrete real
Execute example, and referring to the drawings, the present invention is described in more detail.
The ultra micro planar electrode array sensor that the present invention proposes includes: ultramicroelectrode array and composite membrane
Layer, wherein ultramicroelectrode array by dielectric base, microelectrode array, to electrode, reference electrode, electricity
Pole lead-in wire and contact are constituted.Described dielectric base is the carrier of whole chip, on dielectric base surface
Center, distributed circle that several are arranged in the matrix form, that be made up of conductive film material
Shape ultramicroelectrode, constitutes microelectrode array.Described microelectrode array includes multiple, is distributed in insulation base
, multiple microelectrode arrays can share reference electrode and to electrode at the end.Composite film be nano-modified layer,
Bio-compatible decorative layer, specific bio-layer decorative layer organic assembling form, and its subregion is modified at ultra micro
Electrod-array surface.Ultramicroelectrode be plane be due to noble metal conductive layer sputtering thickness be only
100～300nm.The electrode that this thickness is formed is flat film.Electrod-array in centre position,
Contact is in surrounding, to electrode and reference electrode according to design in the side of electrod-array；Lead-in wire is to connect
Contact and electrod-array.
Wherein, described microelectrode array select the metal that conductive film material is good biocompatibility or
Metallic compound；Microelectrode array comprises 60～128 microelectrodes, and wherein microelectrode is according to its surface
The difference of the composite function film layer modified can be used in the detection of Electrophysiology signal, for neuroelectricity
Microelectrode diameter 0.5 μm of bio-signal acquisition～5 μm, additionally it is possible to believe for neurotransmitter electrochemistry
Number detection, and is used for applying electro photoluminescence, applies microelectrode diameter 0.5 μm～5 μm of electro photoluminescence,
Microelectrode pitch 5 μm～200 μm.
The conductive film material of lead-in wire and contact is identical with microelectrode, and thickness is more than 300nm, it is ensured that its
Mechanical strength can bear the pressure that standard electronic components and parts Elastic metal probe is caused, and ensures
Repeatedly cell is cultivated after detection uses cleaning and is still had good electric conductivity.
Described ultra micro planar array sensor, can detect electricity physiological signal and the electrochemistry letter of nerve cell
Number, the neurotransmitter Quantum neural computing signal of the many synaptic sites of multiple nerve cell can be detected simultaneously；Can be same
Time detect single nerve cell various neurotransmitters signal.
Heretofore described dielectric substrate materials is by the group formed from glass, heat resistant glass, silicon chip etc.
In the one that chooses make.This base material hardness is moderate with thickness, can splash-proofing sputtering metal layer, tolerate one
Determine high temperature, lay the foundation for next step technique.
Fig. 1 shows the preparation method work of a kind of ultra micro planar electrode array sensor that the present invention proposes
Process flow figure.As it is shown in figure 1, the method includes:
Step 1, dielectric base is carried out process after, on substrate coat photoresist；Can adopt
Use positive glue, it would however also be possible to employ negative glue, different photoresist can be used according to difference plate-making mode, use general
Lead-in wire needed for being formed after logical photoetching process photoetching development on a photoresist and electrode, constitute microelectrode battle array
Row pattern.
Step 2, on full wafer base material sputter microelectrode conductive membrane layer, described microelectrode conductive film
Layer is one layer of electrode metal layer, and stripping photoresist leaves required electrode, lead-in wire and contact, basis of formation
Metal electrode array.Electrode is made up of double-layer structure, including lower floor's Seed Layer titanium (Ti) of metal
Or chromium (Cr) and be made up of a kind of noble metal choosing in the group constituting from gold, platinum and palladium
Upper strata.On insulating barrier, titanium or the chromium lower metal layer of pre-sputtering are beneficial to gold or platinum and glue with the effective of electrode surface
Knot.Precious Metals-Gold and platinum are suitable as electrode, because they are in the stability of electrode surface area, electrification
The electrochemical properties of the aspects such as reproducibility, non-oxidizability is the best；And process simple, with
The cementability of glass/silicon is good and conductance high, and has bio-compatibility.The platinum of the present invention or layer gold
It is as thin as 200nm～500nm.
Step 3, in basic electrode array surface, pass through plasma enhanced chemical vapor deposition
(PECVD) one in silica, silicon nitride, silica etc. is as insulating barrier, and its thickness exists
Between 250nm～500nm.
Step 4, there is the first area of basic electrode array of insulating barrier in deposition, use two-layer wiring
Ground floor domain carry out secondary photoetching, use stepping photoetching process to carry out ultramicroelectrode array Centered Graphs
The formation (especially for 0.5 μm～2 μm electrode site diameters) of shape, then uses plasma etching
Method, expose the ultramicroelectrode array A in the first area of basic electrode array and touch accordingly
Point, retains the insulating barrier that all wire surfaces cover；This step purpose is to use insulating barrier to cover
Lead-in wire in basic electrode array, and etch expose the ultramicroelectrode array A being positioned in first area
And corresponding contacts；
Step 5, there is the second area of basic electrode array of insulating barrier in deposition, repeat step 4 quarter
Erosion exposes ultramicroelectrode array B, to electrode, reference electrode and corresponding contacts B.Wherein, double-deck
Wires design can form reference electrode at ground floor, at the second layer, electrode can also be formed reference electricity
Pole, to electrode, ultramicroelectrode array A and B can share reference electrode and to electrode.Layer cloth
Line is under ensureing good conductive wire scenario, it is simple to form more dense ultramicroelectrode in limited plane
Site, is beneficial to capture the Quantum neural computing in synapse cell site.
Step 6, partitioned organization: use PDMS, polyimides, SU-8 etc. to form partitioned organization,
Isolation dam is formed, by multiple electricity with the polymeric material that height is 2 μm～2000 μm between different structure
Pole site separates becomes multiple region, it is simple to different modifying and various neurotransmitters detection, at regional
Carry out not homospecificity to modify and various neurotransmitters detection.Polymeric barrier materials selection PDMS,
Polyimides, SU-8 one or both combining structure therein.
Step 6, forming composite function film layer at electrode face finish, it is modified by following sequencing
Carry out: nano-modified, specific bio-layer is modified, bio-compatible is modified, can root for zones of different
According to needing a layer, two-layer or three layers modifying in above-mentioned three layers.
Nano-modified layer: different subregions fix nano material, in order to increase electrode performance.Nano-modified
Layer uses metal or other excellent conductive material to carry out pointed decoration.Modified metal material include platinum, gold,
Titanium, rhodium, osmium etc., wherein preferred platinum black, titanium nitride.Other excellent conductive material includes carbon, poly-pyrrole
Cough up, preferably carbon nanomaterial.Clear in order to obtain the electrode surface plasma of high-efficiency electrochemical activity
Washing machine carries out activation process to electrod-array.Electrode, after surface activation process, is possible not only to remove
The pollutant of electrode surface, can also play the effect on activated electrode surface simultaneously, increases electrode surface
Hydrophily, the condition of etching can do suitable adjustment according to the situation of electrode surface.Nano-modified layer is fixed
Point modification can use plating, electropolymerization, combined polymerization and micro-coating etc..
The special reaction decorative layer that selects: fixing different specific materials is (such as structure choice sex modification material
The biologic specificity decorative materials etc. such as material, enzyme), for the neurotransmitter that correspondence detection is different.Modify
It is assembled into ultra micro planar array sensor.Special selection reaction decorative layer includes enzyme or antibody or antigen
Or macromolecule M8003 line reagent and electron acceptor, auxiliary reagent.Enzyme reagent includes oxidizing ferment, reduction
Enzyme etc.；Antibody includes monoclonal antibody, how anti-etc.；Electron acceptor include electron acceptor by from ferricyanate,
One selected in the group of methylene blue, Ferrocene and its derivative and rhodium/osmium ion polymer composition
Kind；Auxiliary reagent is made up of coupling reagent, enzyme/antibody activator, buffer solution and surfactant etc..
Bio-compatibility selective modification, including the affine modification in local of ultramicroelectrode site areas, uses
In polylysine (polylysine), mercaptopropionic acid, ethylene imine (PEI), laminine (laminin)
One or combination.Lead areas uses PEG, Teflon etc. to carry out repelling the modification of protein substance.
Wherein, nano-modified layer, special selection reaction decorative layer, bio-compatibility decorative layer are the most suitable
Sequence is modified.Successively can modify at the same area, it is also possible to modify respectively in zones of different.
Embodiments provide a kind of method carrying out nano-modified layer at electrode surface.This enforcement
In example, carry out nano-modified by assembling one layer of nano particle at electrode surface.Concrete, by
Electrode surface plating platinum black or electropolymerization polypyrrole, can obtain the electrode nanometer layer with different pore size.
The method further enhances the hydrophily of electrode surface, increases the effective surface area of electrode.Not only
Be conducive to the immobilization of enzyme reagent, and add the electro-chemical activity of electrode.
Reaction reagent is fixed in electrode nanometer layer formation reagent layer by the present invention, is fixed on electrode nanometer
Reaction reagent on layer includes electron acceptor, reacts with analyte and can produce and analyte
The composite reagent of the coupled reaction of the electric current that concentration is corresponding, the most also includes that electron mediator, enzyme try
Agent, buffer solution and surfactant etc..
The reaction principle of agent combination institute of the present invention foundation is as follows:
Wherein Ach acetylcholine；AchE acetylcholine lipolytic enzyme；Acetate lactic acid；choline
Choline；ChOx choline oxidase, betaine aldehyde trimethylamine acetaldehyde.
Embodiments of the invention, it is provided that the method fixing reaction reagent on nano particle.
First at electrode face finish osmium polymer conductive media body polymer, normal temperature is placed 12 hours
Above.Then by the acetylcholine oxidizing ferment of debita spissitudo, acetylcholine lipolytic enzyme, glutaraldehyde and ox
After the mixing such as seralbumin, it is coated in electrode surface immediately, is dried in 35～37 DEG C of drying boxes
After 20～25 minutes, deionized water rinsing removes uncrosslinked glutaraldehyde, after air drying, seals low
Temperature storage.
Play the electron acceptor of redox by from redox polymers (Os, rhodium etc.), iron
Cyanate, methylene blue, Ferrocene and its derivative, to stupid quinone, phenazine methosulfate, indophenols and
The one selected in the group of its derivative and β-Naphthoquinone-4-potassium Sulfo-nate composition；By electron mediator by enzyme
The electronics produced in course of reaction is from enzyme reaction central transference to electrode surface so that current mode enzyme senses
Response speed and the detection sensitivity of device are improved, and reduce the voltage of reaction simultaneously.Described electricity
Sub-acceptor is Os redox polymers, can effectively reduce detection limit, make operating potential be down to 0V
Left and right, reduces the interference of other active material in blood.
Buffer solution is phosphate buffer, TRIS buffer solution, MES buffer solution and physiological saline composition
The one selected in group；Described buffer solution is phosphate buffer.Buffer solution is for providing a pH
The reaction environment of value stabilization, Optimal pH reaction is 6～8.
Described surfactant is TritonX-100.Interpolation 0.01～the surface-active of 1% nonionic
Agent, improves the affinity rate of mixed liquor and strip, makes mix reagent be easier to be coated uniformly on rapidly electricity
Surface, pole, the coat of formation is uniform and thin, electron transport rate when being beneficial to improve detection.Work as surface
When surfactant concentration is higher than 0.5%, enzymatic activity is suppressed and affects.Therefore 0.01～0.1% is selected
Concentration surface activating agent.
Above-mentioned various purposes, method, feature and the advantage of the present invention, by below in conjunction with the accompanying drawings and real
Execute example and can obtain more detailed explanation.
Fig. 1, Fig. 2 are preparation technology flow chart and the local of the ultramicroelectrode sensor that the present invention designs
Enlarged plan view.First clean insulating substrate sheet glass (thickness about 1mm), be successively coated with on glass
Covering photoresist (positive glue 6130 or 2840 or AZ1500), common photoetching development forms electrode and lead-in wire touches
Dot pattern, then sputtering noble metal electrode film Ti basalis (thick 20～30nm) and Pt
(200～300nm), stripping photoresist, basis of formation electrod-array；PECVD depositing insulating layer
Si3N4 (300～500nm)；Use stepping photoetching process in the first area of basic electrode array surface
Carry out secondary photoetching (the ground floor domain using two-layer wiring is carried out), needed for photoresist is formed
Ultramicroelectrode array figure；Reactive ion etching process is used to perform etching ultramicroelectrode array figure
Except insulating barrier, insulating barrier and electrode material membrane form ultramicroelectrode array figure A and contact
(15 × 4 array of row electrodes: electrode site diameter is respectively 2 μm, 3 μm, 4 μm, 5 μm
(as shown in Figure 3)；The least electrode is more beneficial to vesica release detection)；At basic electrode array
The second area on surface repeats photoetching, noble metal sputtering, covers insulating barrier, uses the of two-layer wiring
Two layers of domain carry out secondary photoetching and plasma etching step, prepare ultramicroelectrode array B, right
Electrode, reference electrode and contact B (15 × 4 array of row electrodes: electrode site diameter be respectively 2 μm,
3 μm, 4 μm, 5 μm (as shown in Figure 3)).Spin coating SU-8 glue, common photoetching development, formed
The electrod-array of cut zone (10～20 μm height segmentation dam).
Ultramicroelectrode array forms nanometer layer (as shown in Figure 4) by plating.Electroplate liquid is 0.1M's
Pyrroles, the KCl of 0.1M, use the plating operating potential of 0.5～0.7V to carry out constant voltage plating, or
The operating current using 0.01～0.05mA carries out constant current plating, and electroplating time is 30 seconds～3 minutes,
-0.1V～0.8V can also be used to be circulated volt-ampere plating, scanning 20～40 circle.Can also select
Au, Pt carry out the plating of nano metal material.Then enzyme layer is modified, such as acetylcholine detection region:
Acetylcholine lipolytic enzyme 100U/ml；Choline oxidase: 100U/ml；Triton X-100:0.01%, penta
Dialdehyde: 0.1%, bovine serum albumin(BSA): 5%.Dopamine detection region then modifies 0.1～1%Nafion, dry
Dry formation Nafion membrane.Different transmitter detection zone modifies different enzymes or specific detection material.Modify
Cheng Hou, seals Cord blood.Before using, first carry out ultraviolet sterilization, then enter in electrode site district
The selective modification of row bio-compatibility material, selects polylysine to be coated or press back.The most again
Inoculation nerve cell is cultivated.
The present invention has used preferred embodiment to illustrate, preferred embodiment for illustrative purposes only,
Rather than limitation of the present invention.The present invention can be made on the basis of the above description many improve and
Change.Therefore, within the scope of the appended claims, the present invention can have is not above-mentioned other
Implementation.Such as: the electricity that the difference of electrode shape, other nano particle and non-nano material are modified
Pole reaction zone, different agent combination forms etc..
Particular embodiments described above, is carried out the purpose of the present invention, technical scheme and beneficial effect
Further describe, be it should be understood that the foregoing is only the present invention specific embodiment and
, be not limited to the present invention, all within the spirit and principles in the present invention, that is done any repaiies
Change, equivalent, improvement etc., should be included within the scope of the present invention.
Claims (9)
1. a ultra micro planar electrode array sensor, it is characterised in that: include ultra micro plane electrode battle array
Row and composite function film layer, wherein said ultra micro planar electrode array by dielectric base, microelectrode array,
Constituting electrode, reference electrode, contact conductor and contact, described microelectrode array includes multiple,
On a dielectric base, multiple microelectrode arrays can share reference electrode and to electrode in distribution；Described compound
Functional film layer includes that nano-modified, bio-compatible is modified, specific bio-layer modifies the decorative layer combined,
Its subregion is modified at ultra micro planar electrode array；
Wherein, described ultra micro planar array sensor can detect the many synaptic sites of multiple nerve cell simultaneously
Neurotransmitter Quantum neural computing signal and/or detect single nerve cell various neurotransmitters signal simultaneously.
2. ultra micro planar electrode array sensor as claimed in claim 1, it is characterised in that: it is adopted
Prepared by the method combined by micro electro mechanical system (MEMS) technology, nano-modified technology and bio-modification technology.
Ultra micro planar electrode array sensor the most according to claim 1, it is characterised in that: institute
State the metal that conductive film material is good biocompatibility or metallic compound that microelectrode array is selected；
Microelectrode array comprises 60～128 microelectrodes, the compound merit that wherein microelectrode is modified according to its surface
Energy film layer is different and has following difference in functionality: Electrophysiology signal detection, neurotransmitter electrochemistry
Signal detection and applying electro photoluminescence.
Ultra micro planar electrode array sensor the most according to claim 1, it is characterised in that: use
In microelectrode diameter 0.5 μm～5 μm of Electrophysiology signal detection, for neurotransmitter electrochemistry
Signal detection and apply microelectrode diameter 0.5 μm of electro photoluminescence～5 μm, microelectrode pitch 5 μm～
200μm；The conductive film material of lead-in wire and contact is identical with microelectrode, and thickness is more than 300nm.
5. the preparation method of a ultra micro planar electrode array sensor, it is characterised in that include as follows
Step:
Step 1, at spin coating one layer photoetching glue in the dielectric base of surface clean, after photoetching development
Form lead-in wire and microelectrode array pattern；
Step 2, microelectrode array patterned surfaces sputter one layer of microelectrode conductive membrane layer；
Step 3, use stripping technology remove unnecessary microelectrode conductive membrane layer, stay required electrode,
Lead-in wire and contact, basis of formation metal electrode array；
Step 4, on underlying metal electrod-array surface by plasma enhanced chemical vapor deposition exhausted
Edge layer；
Step 5, there is the first area of basic electrode array of insulating barrier in deposition, use two-layer wiring
Ground floor domain carry out secondary photoetching, use stepping photoetching process to form ultramicroelectrode array Centered Graphs
Shape, the method then using plasma etching, expose the first ultramicroelectrode array and contact, retain
The insulating barrier that all wire surfaces cover；
Step 6, there is the second area of basic electrode array of insulating barrier in deposition, repeat step 5 quarter
Erosion exposes the second ultramicroelectrode array and corresponding contacts；
Step 7, employing SU-8, PDMS or polyimides preparation have on the chip of ultramicroelectrode array
By being commonly lithographically formed dam type partitioned organization.
Method the most according to claim 5, it is characterised in that expose the first ultra micro electricity in etching
Reference electrode is formed and to electrode, described first ultramicroelectrode when pole array or the second ultramicroelectrode array
Array and the public reference electrode of the second ultramicroelectrode array and to electrode.
Method the most according to claim 5, it is characterised in that described dam type partitioned organization height
Being 2 μm～2000 μm, it is by multiple electrode site are separated into multiple region, with at each
Region carries out not homospecificity and modifies and various neurotransmitters detection.
Method the most according to claim 5, it is characterised in that described method also includes:
Step 8, on each dam type partitioned organization modification complex function film layer, it is nano-modified, raw
Thing special layer is modified, bio-compatible is modified to combine and is prepared from.
Method the most according to claim 8, it is characterised in that: step 8 specifically includes:
Step 81, modifying nano-modified layer, nano-modified layer uses metal or other excellent conductive material
Carrying out pointed decoration, its modification mode uses plating, electropolymerization, combined polymerization and micro-coating；
Step 82, modifying the special reaction decorative layer that selects, special selection reaction decorative layer includes enzyme or anti-
Body or antigen or macromolecule M8003 line reagent and electron acceptor, auxiliary reagent；
Step 83, bio-compatibility selective modification, repair including ultramicroelectrode site areas local is affine
Decorations, use in polylysine, mercaptopropionic acid, ethylene imine (PEI), laminine (laminin)
One or more combination, lead areas uses PEG, Teflon to carry out repelling repairing of protein substance absorption
Decorations.
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