CN108072687A - It is a kind of that the method without enzyme biologic sensor is prepared based on microelectrode array - Google Patents
It is a kind of that the method without enzyme biologic sensor is prepared based on microelectrode array Download PDFInfo
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Abstract
The invention discloses it is a kind of prepare microelectrode array based on optical etching technology and modify preparation method of the electrochemistry without enzyme sensor is prepared with conductive element and inorganic sensitive coating, be related to the fields such as polymer material science, optical etching technology, electrochemical sensor.The photoresist used in the present invention can realize the preparation of superelevation draw ratio microelectrode array, three-dimensional structure is developed on the basis of two-dimension plane structure, greatly increase the specific surface area of coating, it lays a good foundation to prepare highly sensitive electrochemical sensor, the micro array structure of 3 D stereo improves the roughness of coating surface simultaneously, is conducive to the payload of sensitive coating.The method of the electrochemical in-situ reduction inorganic transition metal nano-particle used in the present invention realizes the preparation of the inorganic sensitive coating with electrochemical response, has synergistic effect with conductive microelectrode array.The technology used in the present invention is easy to operate, and constructed sensor has many advantages, such as that high sensitivity, stability are good for analysans.
Description
【Technical field】
This patent is related to electrochemical sensor field and photoengraving field, more particularly, to one kind with photoetching microelectrode array
For substrate modified conducting layer and the electrode of inorganic sensitive coating, and the method for being prepared into no enzyme biologic sensor.
【Background technology】
In recent years, more serious the problems such as aging of population, chronic disease, the early detection of disease and intervention seem particularly heavy
Will, at the same time, the research of various kinds of sensors part becomes hot spot, wherein being concentrated the most with the research of electrochemical sensor.
Electrochemical sensor be it is a kind of bio signal is changed into electric signal, detect material concentration and other biological respond
Analytical equipment.It is usually made of recognition component, energy converter and signal amplifying apparatus, electrode believes concentration as conversion element
Number measurable electric signal signal in response is converted into, realizes the quantitative or qualitative analysis to target analytes.Traditional
Electrode is made of two-dimension plane structure, and specific surface area is small, the poor adhesive force with recognition component, causes the senser element of structure
Sensitivity is relatively low, and current-responsive is smaller, seriously constrains the development of sensor.For this purpose, researchers have carried out multiple exploration, hair
The specific surface area for now improving electrode material and the i.e. raising electrode material of active site of detectable substance is effective method the most direct,
Therefore, they propose array structure of the nano-particle using zero dimension, one-dimensional nano wire, the nanometer sheet of two dimension and three-dimensional etc.
Serial of methods, most effective one are the improved methods in 3D structures.
The method for increasing specific surface area in 3D structures usually prepares porous structure, micro array structure and composite microstructure
Deng these methods can significantly improve sensitivity and the current-responsive of sensor, but the system of porous structure and composite microstructure
Standby to have uncontrollability, lot-to-lot variability is big, be unfavorable for it is commercialized be widely applied, and micro array structure just presents controllably
Property strong, advantage that lot-to-lot variability is small, while also there is high response current, the advantages that high s/n ratio, fast-response rate, this is it
His all incomparable advantage of structure.The method for preparing microelectrode array usually has chemical vapour deposition technique, photoetch method and water
Hot method etc., wherein most strong with the method design of photoengraving, controllability is most strong, and required reaction condition is most mild, can be by micro- electricity
The advantage of this body structure of pole array expands to maximum.
Photoengraving refers under illumination effect, by photoresist by the mistake in the pattern transfer on mask plate to substrate
Journey.The part of wherein most marrow is exactly photoresist used, can be divided into positive photoresist and negativity according to the difference of reaction mechanism
The characteristics of the characteristics of photoresist, positive photoresist is that exposed portion is washed away and leaves backlight portion, negative photoresist is exposure portion
Divide and crosslink to be left.Photoengraving is also applied in sensory field more and more in recent years, representative device
It is exactly interdigital electrode, although interdigital electrode can also increase the specific surface area of electrode material to a certain extent, has very strong
Limitation.
Therefore, the microelectrode array of high-specific surface area is prepared using the method for photoengraving and is applied in electrochemical sensing,
Exploitation is of great significance with highly sensitive and fast-response time electrochemical sensor.
【The content of the invention】
In view of the above-mentioned problems existing in the prior art, the present invention is intended to provide one kind prepares microelectrode based on optical etching technology
Array is simultaneously modified with conductive element and prepares method of the electrochemistry without enzyme sensor without enzyme sensitive coating.What is used in the present invention is negative
Property photoresist can realize the preparation of high length-diameter ratio microelectrode array, and three-dimensional structure is developed on the basis of two-dimension plane structure,
The specific surface area of coating is greatly increased, the electrochemical sensor to prepare highly sensitive is laid a good foundation, while 3 D stereo
Micro array structure improves the roughness of coating surface to a certain extent, is conducive to the payload of electrode surface.The present invention
The method of the electrochemical in-situ reduction inorganic transition metal nano-particle of middle use realizes the inorganic biography with electrochemical response
Feel the preparation of coating, there is synergistic effect with conductive microelectrode array, the advantage of high-specific surface area is further expanded.The present invention
Used technical operation is easy, and constructed sensor has many advantages, such as that high sensitivity, stability are good for analysans.
Technical scheme is as follows：
It is a kind of that the method without enzyme biologic sensor, the preparation of photoetching microelectrode array, conduction are prepared based on microelectrode array
The structure of preparation and electrochemistry without enzyme sensor of layer is as follows：
(1) preparation of photoetching microelectrode array
One layer of tackifier of spin coating after pretreating substrates coat on negative photoresist after dry, carry out front baking, exposure,
It dries, develop, post bake technique afterwards, complete, by the process of pattern transfer on mask plate to substrate surface, to obtain required array of figure
Case.
The base material for silicon chip, glass plate, aluminium flake, copper sheet it is any, need to be pre-processed before use.Processing method：
Substrate surface is cleaned using acetone, removes surface and oil contaminant and impurity, the base material cleaned then is positioned over temperature is
30min is toasted in 200 DEG C of thermal station to remove the solvent of substrate surface.
The array has the specific surface area of superelevation, therefore negative photoresist selection can prepare high aspect ratio micro-nano structure
2000 series 2025,2035,2050, any one of 2075 of SU-8 Microchem, tackifier OMNICOAT；
Figure is the array of circular apertures being distributed in regular hexagon on the mask plate, and Circularhole diameter is 3 μm~50 μm, between circular hole
It is light transmission part away from equal with diameter, inside circular hole.
The technological parameter of the photolithographic process is as follows：
Film：Film is carried out using any one in spin coater, blade applicator, spreading rod, wet-film thickness is 20 μm~120
μm；
Front baking：Photoresist film is placed in 65 DEG C of baking ovens and keeps 1min, 95 DEG C of 5~15min of maintenance is warming up to, then cools down
To 65 DEG C of maintenance 1min, 15min is stood；
Exposure：Wavelength is 365nm, light intensity 12.9mw/cm2, the time is 5s~30s；
After dry：1min is kept in 65 DEG C of baking ovens, is warming up to 95 DEG C of 5~15min of maintenance, cooling and standings；
Development：It is placed in development 5s~2min in propylene glycol methyl ether acetate (PGMEA)；
Post bake：5~10min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
Conductive element is introduced on the basis of (1), uniform conductive layer is formed in microarray surface；
The method for introducing conductive element, except vacuum vapor plating, vacuum sputtering coating, vacuum ion membrane plating and chemistry
Vapor deposition is outer, also forms interlayer using dopamine (DA) adhesiveness, then forms conduction by suction-operated adsorbing metal
The indirect method of layer；
The conductive element is golden (Au), silver (Ag), copper (Cu), nickel (Ni), titanium (Ti), platinum (Pt), carbon nanotubes
(CNT), one kind in graphene (GN)；
The conductive layer thickness is 20nm~80nm.
(3) structure of the electrochemistry without enzyme sensor
The inorganic sensitive coating with electrochemical response is prepared on the basis of (2), first by above-mentioned conductive microelectrode battle array
Row are immersed in inorganic transition metal solion, and connection calomel electrode, platinum electrode form three-electrode system, in electrochemical operation
Apply the I-t deposition procedures that appropriate voltage carries out certain time in standing, inorganic transition metal ion is reverted in situ conductive micro-
Electrod-array surface.Ultrapure water and logical nitrogen (N during taking-up2) dry.Obtain with highly sensitive electrochemistry without
Enzyme sensor.
The inorganic transition metal solion can be golden (Au), silver-colored (Ag), copper (Cu), nickel (Ni), zinc (Zn), platinum
(Pt), any one in the solion of cobalt (Co), palladium (Pd), cadmium (Cd), concentration are 5mM~10mM.
The voltage that the deposition process applies is -0.5V~-2.0V, and the time is 30s~200s.
The present invention is beneficial to be had technical effect that：
1st, the present invention is prepared for patterned microelectrode array using the method for photoengraving in substrate surface, by the two of plane
Dimension structure updating is three-dimensional structure, considerably increases the specific surface area and surface roughness of coating.Meanwhile microelectrode in array
Parallel connection also can effectively amplify response current, reduce the response time, enhance signal transmission and increase signal-to-noise ratio；
2nd, present invention introduces conductive element, can the signal intensity of sensing interface be effectively transferred to analysis system, had
Beneficial to the senser element for constructing highly sensitive, high stability；
3rd, the present invention is realized using the method for electrochemical in-situ reduction inorganic transition metal nano-particle with electrochemistry
The inorganic sensitive coating of response has with the conductive microelectrode array of three-dimensional structure and cooperates with amplified signal in the load of electrode surface
Effect, prepared sensor have excellent stability and detection performance；
4th, the electrochemistry for preparing of the present invention without enzyme sensor prepare with it is easy to operate, the radiation of traditional detection method can be avoided
Harm, background noise are big, very complicated, the shortcomings of analysis time is long, instrument is valuable and needs professional operator.
5th, the present invention, which is combined optical etching technology with electrochemical sensing technology, can build new, diversified electrochemistry without enzyme biography
Sensor is expected to be widely applied to the fields such as food security, biological medicine and environment monitoring.
6th, electrochemistry of the present invention without enzyme sensor can realize to food additives, environmental contaminants it is online, real-time, fast
Speed, accurate detection, have extensive research and actual application value.
【Description of the drawings】
Fig. 1：The preparation method schematic diagram of the conductive microelectrode array of the present invention；
Fig. 2：The super depth-of-field microscope figure of microelectrode array is made in the embodiment of the present invention 1；
Fig. 3：The super depth-of-field microscope figure of microelectrode array is made in the embodiment of the present invention 2；
Fig. 4：Cyclic voltammetry curve of the electrochemistry without enzyme sensor detection hydrogen peroxide is made in the embodiment of the present invention 3；
【Specific embodiment】
With reference to the accompanying drawings and examples, the present invention is specifically described.It should be appreciated that following embodiment is only this hair
Bright preferred embodiment to more fully understand the present invention, thus should not be taken as limiting the scope of the invention.
Embodiment 1
(1) preparation of photoetching microelectrode array
One layer of tackifier (OMNICOAT) of spin coating after silicon chip is pre-processed, by negative photoresist SU-8 after drying
2025 coat on, and carry out front baking, exposure, rear baking, development, post bake technique, complete pattern transfer on mask plate to base material table
The process in face obtains required array pattern.Photoetching process is：
Film：Spin-coating method, 10s under forward 600r/min, after turn 40s under 2600r/min, wet-film thickness is 50 μm；
Front baking：It is placed in 65 DEG C of baking ovens and keeps 1min, be warming up to 95 DEG C of maintenance 6min, be then cooled to 65 DEG C of maintenances
1min stands 15min；
Exposure：Circularhole diameter and spacing specification are selected as 50 μm of mask plate, exposure wavelength 365nm, light intensity 12.9
mw/cm2, time 9s；
After dry：2min is kept in 65 DEG C of baking ovens, is warming up to 95 DEG C of maintenance 6min, cooling and standings；
Development：It is placed in development 30s in propylene glycol methyl ether acetate (PGMEA)；
Post bake：10min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
(thickness about 50nm) Au layers uniform is formed in microarray surface using vacuum sputtering coating on the basis of (1),
120 DEG C are then heated in an oven, are kept 80min, are cooled to room temperature；
(3) structure of the electrochemistry without enzyme sensor
The inorganic sensitive coating with electrochemical response is prepared on the basis of (2), first by above-mentioned conductive microelectrode battle array
Row immerse the copper sulphate (CuSO of 5mM4) in solution, connection calomel electrode, platinum electrode form three-electrode system, in electrochemistry
Using the deposition voltage deposition 80s of -0.8V in work station, by copper nano-particle (CuNPs) in-situ reducing to conductive microelectrode battle array
List face.Ultrapure water and logical nitrogen (N during taking-up2) dry.It obtains to hydrogen peroxide (H2O2) there is highly sensitive detection
The electrochemistry of property is without enzyme sensor.
Embodiment 2
(1) preparation of photoetching microelectrode array
One layer of tackifier (OMNICOAT) of spin coating after silicon chip is pre-processed, by negative photoresist SU-8 after drying
2025 coat on, and carry out front baking, exposure, rear baking, development, post bake technique, complete pattern transfer on mask plate to base material table
The process in face obtains required array pattern.Photoetching process is：
Film：Spin-coating method, 10s under forward 600r/min, after turn 40s under 4350r/min, wet-film thickness is 20 μm；
Front baking：Photoresist film is placed in 65 DEG C of baking ovens and keeps 1min, 95 DEG C of maintenance 5min is warming up to, is then cooled to 65
DEG C maintain 1min, stand 15min；
Exposure：Circularhole diameter and spacing specification are selected as 10 μm of mask plate, exposure wavelength 365nm, light intensity 12.9
mw/cm2, time 6s；
After dry：1min is kept in 65 DEG C of baking ovens, is warming up to 95 DEG C of maintenance 5min, cooling and standings；
Development：It is placed in development 15s in propylene glycol methyl ether acetate (PGMEA)；
Post bake：10min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
(thickness about 50nm) Au layers uniform is formed in microarray surface using vacuum sputtering coating on the basis of (1),
120 DEG C are then heated in an oven, are kept 80min, are cooled to room temperature；
(3) structure of the electrochemistry without enzyme sensor
The inorganic sensitive coating with electrochemical response is prepared on the basis of (2), first by above-mentioned conductive microelectrode battle array
Row immerse the copper sulphate (CuSO of 5mM4) in solution, connection calomel electrode, platinum electrode form three-electrode system, in electrochemistry
Using the deposition voltage deposition 80s of -0.8V in work station, by copper nano-particle (CuNPs) in-situ reducing to conductive microelectrode battle array
List face.Ultrapure water and logical nitrogen (N during taking-up2) dry.It obtains to hydrogen peroxide (H2O2) there is highly sensitive detection
The electrochemistry of property is without enzyme sensor.
Embodiment 3
(1) preparation of photoetching microelectrode array
One layer of tackifier (OMNICOAT) of spin coating after silicon chip is pre-processed, by negative photoresist SU-8 after drying
2050 coat on, and carry out front baking, exposure, rear baking, development, post bake technique, complete pattern transfer on mask plate to base material table
The process in face obtains required array pattern.Photoetching process is：
Film：Spin-coating method, 10s under forward 600r/min, after turn 40s under 1000r/min, wet-film thickness is 100 μm；
Front baking：Photoresist film is placed in 65 DEG C of baking ovens and keeps 5min, 95 DEG C of maintenance 10min is warming up to, is then cooled to
65 DEG C of maintenance 1min, stand 15min；
Exposure：Circularhole diameter and spacing specification are selected as 5 μm of mask plate, exposure wavelength 365nm, light intensity 12.9
mw/cm2, time 11s；
After dry：2min is kept in 65 DEG C of baking ovens, 95 DEG C is warming up to and maintains 8min, cooling and standings 1min；
Development：It is placed in development 30s in propylene glycol methyl ether acetate (PGMEA)；
Post bake：15min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
(thickness about 50nm) Au layers uniform is formed in microarray surface using vacuum sputtering coating on the basis of (1),
120 DEG C are then heated in an oven, are kept 80min, are cooled to room temperature；
(3) structure of the electrochemistry without enzyme sensor
The inorganic sensitive coating with electrochemical response is prepared on the basis of (2), first by above-mentioned conductive microelectrode battle array
Row immerse the copper sulphate (CuSO of 5mM4) in solution, connection calomel electrode, platinum electrode form three-electrode system, in electrochemistry
Using the deposition voltage deposition 80s of -0.8V in work station, by copper nano-particle (CuNPs) in-situ reducing to conductive microelectrode battle array
List face.Ultrapure water and logical nitrogen (N during taking-up2) dry.It obtains to hydrogen peroxide (H2O2) there is highly sensitive detection
The electrochemistry of property is without enzyme sensor.
Test case
Electrochemistry is prepared without enzyme sensor for dioxygen based on photoetching microelectrode array electrochemical reduction inorganic transition metal
The Electrochemical Detection of water.
Electrochemistry prepared by embodiment 3 is immersed without enzyme sensor and the conductive microelectrode array before deposition CuNPs containing 1M
Hydrogen peroxide (H2O2) 0.01M phosphate buffers (pH 7.0) in, using the electrochemistry without enzyme sensor as working electrode, saturation
Calomel electrode is reference electrode, and platinum electrode is to electrode, using Xun Huan volt on Shanghai Chen Hua CHI660A electrochemical workstations
Peace method (CV) is measured.
By test electrochemistry without enzyme sensor deposit CuNPs before and after in hydrogen peroxide (H2O2) phosphate buffer (pH
7.0) the current-responsive variation in can obtain cyclic voltammetry curve comparison diagram, so that it is determined that whether the sensor has to double
The detection of oxygen water.The results are shown in Figure 4, and as seen from Figure 4, prepared electrochemistry has hydrogen peroxide without enzyme sensor
Good current-responsive.
Claims (9)
1. a kind of prepare the method without enzyme biologic sensor, the preparation of photoetching microelectrode array, conductive layer based on microelectrode array
The structure of preparation and electrochemistry without enzyme sensor be as follows：
(1) preparation of photoetching microelectrode array
One layer of tackifier of spin coating after pretreating substrates coat on negative photoresist after dry, carry out front baking, exposure, rear baking,
Development, post bake technique, complete, by the process of pattern transfer on mask plate to substrate surface, to obtain required array pattern.
(2) preparation of conductive layer
Conductive element is introduced on the basis of (1), uniform conductive layer is formed on microelectrode array surface.
(3) structure of the electrochemistry without enzyme sensor
The inorganic sensitive coating with electrochemical response is prepared on the basis of (2), first soaks above-mentioned conductive microelectrode array
Enter in inorganic transition metal solion, connection calomel electrode, platinum electrode form three-electrode system, in electrochemical workstation
Apply the I-t deposition procedures that appropriate voltage carries out certain time, inorganic transition metal ion is reverted into conductive microelectrode in situ
Array surface, ultrapure water and logical nitrogen (N during taking-up2) dry passed to get to highly sensitive electrochemistry without enzyme
Sensor.
2. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the base material for silicon chip, glass plate, aluminium
Any one of piece, copper sheet need to be pre-processed before use, and processing method is：Substrate surface is cleaned using acetone, is gone
Except surface and oil contaminant and impurity, then the base material cleaned is positioned in the thermal station that temperature is 200 DEG C and toasts 30min to remove base
The solvent on material surface.
3. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the array have superelevation specific surface
Product, wherein negative photoresist selection can prepare high aspect ratio micro-nano structure 2000 series 2025 of SU-8 Microchem,
2035th, 2050, any one of 2075, tackifier are OMNICOAT (>=17nm).
4. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the technological parameter of the photolithographic process
For：
(1) film：Any one in spin-coating method, blade applicator method, spreading rod method, wet-film thickness are 10 μm~200 μm；
(2) front baking：Photoresist film is placed in 65 DEG C of baking ovens and keeps 1min, 95 DEG C of 5~15min of maintenance is warming up to, then cools down
To 65 DEG C of maintenance 1min, 15min is stood；
(3) expose：Wavelength is 365nm, light intensity 12.9mw/cm2, the time is 5s~30s；
(4) dry afterwards：1min is kept in 65 DEG C of baking ovens, is warming up to 95 DEG C of 5~15min of maintenance, cooling and standings；
(5) develop：It is placed in development 5s~2min in propylene glycol methyl ether acetate (PGMEA)；
(6) post bake：5~10min in 150 DEG C of baking ovens.
5. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that mask plate used in the photolithographic process
Upper figure is the array of circular apertures being distributed in regular hexagon, and Circularhole diameter is 3 μm~50 μm, and circular hole spacing is identical with diameter, circular hole
Inside is light transmission part.
6. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that it is described introduce conductive element method, remove
Outside these conventional methods of vacuum vapor plating, vacuum sputtering coating, vacuum ion membrane plating and chemical vapor deposition, between can also be used
Connection such as forms interlayer using dopamine (DA) adhesiveness, then the method for conductive layer is formed by suction-operated adsorbing metal.
7. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the conductive element is golden (Au), silver
(Ag), one kind in copper (Cu), nickel (Ni), titanium (Ti), platinum (Pt), carbon nanotubes (CNT), graphene (GN)；The conduction
Layer thickness is 20nm~80nm.
8. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the inorganic transition metal solion
Can be that golden (Au), silver-colored (Ag), copper (Cu), nickel (Ni), zinc (Zn), platinum (Pt), cobalt (Co), palladium (Pd), the ion of cadmium (Cd) are molten
Any one in liquid, concentration are 5mM~10mM.
9. according to prepared by claim 1 without enzyme biologic sensor, it is characterised in that the voltage that the deposition process applies for-
0.5V~-2.0V, time are 30s~200s.
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| CN112237682A (en) * | 2019-07-17 | 2021-01-19 | 杭州暖芯迦电子科技有限公司 | Miniature high-density array free-form surface electrode and manufacturing method thereof |
| CN112683975A (en) * | 2020-12-18 | 2021-04-20 | 天津理工大学 | Interdigital microelectrode array electrochemical sensor, preparation method, application and special test box |
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| CN114134528A (en) * | 2021-10-21 | 2022-03-04 | 中国空间技术研究院 | Micron array electrode structure, preparation method and application thereof |
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| CN110838425A (en) * | 2019-12-07 | 2020-02-25 | 福建工程学院 | Three-dimensional carbon array cathode structure modified by metal titanium and preparation method thereof |
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