CN108152350A - A kind of method that enzyme biologic sensor is prepared based on microelectrode array - Google Patents
A kind of method that enzyme biologic sensor is prepared based on microelectrode array Download PDFInfo
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Abstract
The preparation method that microelectrode array and modifying prepares electrochemical enzymatic biosensor with conductive element and molecular recognition sensitive coating is prepared based on optical etching technology the invention discloses a kind of, is related to the fields such as polymer material science, optical etching technology, biological immune technology, electrochemical sensor.The photoresist used in the present invention can realize the preparation of superelevation draw ratio microelectrode array, and three-dimensional structure is developed on the basis of two-dimension plane structure, greatly increase the specific surface area of coating, and the biosensor to prepare highly sensitive provides good microenvironment.The method of covalent fixed member Recognition unit enzyme used in the present invention realizes effective fixation of enzyme, and the advantage of high-specific surface area is further expanded, and constructed sensor has many advantages, such as that high sensitivity, high specificity, stability are good for analysans.The combination of optical etching technology and electrochemical sensor can be widely applied to immunoassay and expand be applied to the fields such as food security, biological medicine and environment monitoring.
Description
【Technical field】
This patent is related to electrochemical sensor field and photoengraving field, more particularly, to one kind with photoetching microelectrode array
The electrode of sensitive coating is identified for substrate modified conducting layer and specific molecular, and be prepared into electrochemical enzymatic biosensor
Method.
【Background technology】
In recent years, it is continuous to be faced with aging of population, prevention and treatment of chronic diseases and health medical treatment cost for Global Health medical system
The problems such as increase.For this purpose, researchers realize that disease early detection is efficiently managed with intervention, disease and patient in positive seek
Solution and new technology, reduce cost while health medical treatment service quality is improved.Traditional high-performance Medical Devices
Though the accurate analyzing test data of energy, often there are the problems such as expensive, the testing time is long, is unfavorable for the simple of daily life
Judge.And though the mobile electronic device risen at present can quickly analyze data, there are detection range it is narrow, detection
The problems such as lower limit is insufficient seriously constrains the universal of high performance electronics.Therefore, preparing has wide detection range, low detection
Lower limit, high sensitivity and low-cost electronic device are of great significance.
Electrochemical sensor be it is a kind of bio signal is converted to electric signal, detect material concentration and other biological respond
Analytical equipment.Concentration signal is converted into measurable electric signal signal in response, realization pair by electrode as conversion element
The quantitative or qualitative analysis of target analytes.Compared with other methods, this method has many advantages, such as Miniaturized,
High sensitivity etc., and can be detected in complex sample.
It is the sensitivity for improving sensor that it is most important, which to build electrochemica biological sensor, has numerous researchs in recent years
Persons have made intensive studies it.The result shows that improving the specific surface area of sensitive coating and the signal transmission of coating surface is
Maximally efficient two methods, such as design, improve sensor microstructure prepare microelectrode (Microelectrode, ME) and
Introduce conductive element etc..Microelectrode is that the characteristic size of electrode is less than a kind of electrode of stable state thickness of diffusion layer, usually nanometer
To micron order.Relative to traditional electrode, ME electrodes have high mass transfer rate, high current density, high s/n ratio, Minimum-time constant
The advantages that, the sensitivity of Electrochemical Detection can be effectively improved.However the absolute current of single microelectrode is smaller, general instrument is difficult to
Detection limits its application in analysis detects.Branched microelectrode is composed in parallel into microelectrode array (Microelectrode
Arrays, MEA), it can effectively amplify response current, therefore the system of microelectrode array while single microelectrode advantage is kept
Standby and application has become one of hot spot of analytical chemistry research.In the preparation process of MEA, the orderly of microelectrode array is controlled
Property, microelectrode size, microelectrode spacing are most important for the electrochemical properties of microelectrode array.
Photoengraving is a kind of effective ways for preparing microelectrode array, it refers in the case where illumination acts on, by photoresist
Agent (also known as photoresist) is by the process in the pattern transfer to substrate on mask plate.Its main process is：First, ultraviolet light passes through
Mask plate is irradiated to the substrate surface with one layer of photoresist film, and the photoresist of exposure area is caused to chemically react；With
Afterwards, removal exposure area or the photoresist of unexposed area are dissolved by developing technique (the former claims positive photoresist, and the latter claims to bear
Property photoresist), the figure on mask plate is made to be copied on photoresist film；Finally, pattern transfer is arrived using lithographic technique
On substrate.The shape to form figure, size can be accurately controlled using the method for photoengraving, there is controllability, precision
The advantages that high.At present existing researcher is prepared for the base material of a variety of microscopic patterns using the method for photoengraving, it was demonstrated that photoetching
The feasibility of microscopic pattern is realized, however, the microelectrode array being lithographically derived is applied to highly sensitive electrochemical enzymatic biology
The preparation of sensor has not been reported.
【Invention content】
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 simultaneously modifies the method for preparing electrochemical enzymatic biosensor with conductive element and molecular recognition sensitive coating.It is adopted in the present invention
Negative photoresist can realize the preparation of superelevation draw ratio microelectrode array, and three are developed on the basis of two-dimension plane structure
Structure is tieed up, considerably increases the specific surface area of coating, the biosensor to prepare highly sensitive provides good microenvironment.
The method of covalent fixed member Recognition unit enzyme used in the present invention realizes effective fixation of enzyme, by the excellent of high-specific surface area
Gesture further expands.The technology used in the present invention is easy to operate, and constructed sensor has sensitivity for analysans
The advantages that height, high specificity, stability are good.
Technical scheme is as follows：
A kind of method that enzyme biologic sensor is prepared based on microelectrode array, the preparation of photoetching microelectrode array, conductive layer
Preparation and the structure of enzyme biologic 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,
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 identical with diameter, inside circular hole.
The technological parameter of the photolithographic process is as follows：
Film：Using any one in spin-coating method, blade applicator method, spreading rod method, wet-film thickness is 20 μm~150 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 on microelectrode array surface；
The method for introducing conductive element, except vacuum vapor plating, vacuum sputtering coating, vacuum ion membrane plating and chemistry
It is vapor-deposited outside these conventional methods, also some indirect methods, such as forms middle layer using dopamine (DA) adhesiveness, then lead to
Cross the method that suction-operated adsorbing metal forms conductive layer；
The conductive element is golden (Au), silver (Ag), copper (Cu), nickel (Ni), titanium (Ti), platinum (Pt), carbon nanotube
(CNT), any one of graphene (GN)；
The conductive layer thickness is 20nm~80nm.
(3) structure of enzyme biologic sensor
The bioactive layer with specific molecular recognition capability is prepared on the basis of (2), is led first by prepared by (2)
Electric microelectrode array, which is dipped in the midbody solution with connection function, to be impregnated, with ultrapure water and with nitrogen (N after taking-up2)
Drying.It is subsequently placed into the enzyme solutions of buffer solution configuration, with enzyme dehydrating condensation can occur for the intermediate that early period is covalently attached
Reaction, so as to fulfill effective fixation of enzyme, with phosphate buffer solution (PBS) and ultrapure water and logical nitrogen (N during taking-up2) dry
It is dry.Obtain the enzyme biologic sensor with specific detection type.
The intermediate with connection function can be Mercaptoundecanoic acid (MUA), 4- mercaptobenzoic acids (MBA) and penta
It is any in dialdehyde；A concentration of 5mM~50mM, solvent are ethanol solution, and processing time is 10h~for 24 hours；
In the enzyme solutions, solvent is the phosphate buffer solution of pH 7.0, and enzyme is horseradish peroxidase (HRP), lactic acid
Oxidizing ferment (LOx), glucose oxidase (GOx), cholesterol enzyme, lipase, laccase, tyrosinase, cholinesterase, organophosphor water
It solves one or more in enzyme, nitrate reductase, nitrite reductase, perchlorate reduction's enzyme, urase, carbonic anhydrase.Enzyme
A concentration of 1mg/mL~20mg/mL.
It needs to add in the n-hydroxysuccinimide (NHS) of 15mM and 1- (the 3- dimethylaminos third of 30mM in the enzyme solutions
Base) -3- ethyl-carbodiimide hydrochlorides (EDC), to activated carboxyl, accelerate reaction process.
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 of coating.Meanwhile the in parallel of microelectrode also can be effective in array
Amplify response current, enhance signal transmission；
2nd, present invention introduces conductive elements, can the signal intensity of sensing interface effectively be transferred to analysis system, had
Conducive to the senser element for constructing highly sensitive, high stability；
3rd, the present invention covalently bound method immobilization specific molecular Recognition unit enzyme of use is, it can be achieved that enzyme is effectively consolidated
Fixed and high-efficient carrier, avoids the leakage of enzyme, and prepared sensor has excellent stability and detection performance；
4th, the enzyme biologic sensor for preparing of the present invention prepare with it is easy to operate, the radiation of traditional detection method can be avoided to endanger
The shortcomings of evil, background noise are big, very complicated, 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 novel, diversified electrochemical sensing
Device is expected to be widely applied to the fields such as food security, biological medicine and environment monitoring.
6th, enzyme biologic sensor of the present invention can realize to food additives, environmental contaminants it is online, real-time, quick,
Accurate detection has 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 scanning electron microscope (SEM) photograph of microelectrode array is made in the embodiment of the present invention 1；
Fig. 3：The super depth-of-field microscope of microelectrode array is made in the embodiment of the present invention 2；
Fig. 4：The linear relationship chart of molecular-recognition biosensors 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 15s in propylene glycol methyl ether acetate (PGMEA)；
Post bake：10min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
Au layers uniform (thickness is formed on microelectrode array surface using the method for vacuum sputtering coating on the basis of (1)
Spend about 50nm), it is then heated to 120 DEG C in an oven, keeps 80min, is cooled to room temperature；
(3) structure of enzyme biologic sensor
The bioactive layer with specific molecular recognition capability is prepared on the basis of (2), is led first by prepared by (2)
Electric microelectrode array, which is dipped in 5mM Mercaptoundecanoic acids (MUA), to be impregnated for 24 hours, makes conductive layer that chemical reaction shape occur with intermediate
Into covalent bond, with ultrapure water and with nitrogen (N after taking-up2) drying.The horseradish peroxidase for being subsequently placed into PBS preparations is molten
In liquid, with PBS and ultrapure water and logical nitrogen (N after taking-up2) dry.Obtain the enzyme bio-sensing with specific detection
Device.
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 30 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 20s in propylene glycol methyl ether acetate (PGMEA)；
Post bake：10min in 150 DEG C of baking ovens.
(2) preparation of conductive layer
Form using vacuum sputtering coating that Au layers uniform (thickness is about on microelectrode array surface on the basis of (1)
50nm), it is then heated to 120 DEG C in an oven, keeps 80min, is cooled to room temperature；
(3) structure of enzyme biologic sensor
The bioactive layer with specific molecular recognition capability is prepared on the basis of (2), is led first by prepared by (2)
Electric microelectrode array, which is dipped in 5mM Mercaptoundecanoic acids (MUA), to be impregnated for 24 hours, makes conductive layer that chemical reaction shape occur with intermediate
Into covalent bond, with ultrapure water and with nitrogen (N after taking-up2) drying.It is subsequently placed into the lactate oxidase solution of PBS preparations
In, with PBS and ultrapure water and logical nitrogen (N after taking-up2) dry.Obtain the enzyme bio-sensing with specific detection
Device.
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
Form using vacuum sputtering coating that Au layers uniform (thickness is about on microelectrode array surface on the basis of (1)
50nm), it is then heated to 120 DEG C in an oven, keeps 80min, is cooled to room temperature；
(3) structure of enzyme biologic sensor
The bioactive layer with specific molecular recognition capability is prepared on the basis of (2), is led first by prepared by (2)
Electric microelectrode array, which is dipped in 5mM Mercaptoundecanoic acids (MUA), to be impregnated for 24 hours, makes conductive layer that chemical reaction shape occur with intermediate
Into covalent bond, with ultrapure water and with nitrogen (N after taking-up2) drying.It is subsequently placed into the glucose oxidase solution of PBS preparations
In, with PBS and ultrapure water and logical nitrogen (N after taking-up2) dry.Obtain the enzyme sensor with specific detection.
Test case
The electrochemical enzymatic biosensor of horseradish peroxidase preparation is fixed for dioxygen based on photoetching microelectrode array
The Electrochemical Detection of water.
The phosphate that the biosensor that embodiment 1 prepares is immersed to 0.01M delays in solution (pH 7.4), with every drop 20
The hydrogen peroxide solution of 0.01mM is added dropwise in phosphate buffer by μ L, using the enzyme biologic sensor as working electrode, is satisfied
It is reference electrode with calomel electrode, platinum electrode is to electrode, on Shanghai Chen Hua CHI660A electrochemical workstations, using meter
When current method be measured.
Corresponding linear relationship curve is can obtain with the variation of hydrogen peroxide dripping quantity by testing catalysis oxidation electric current, from
And realize the detection to hydrogen peroxide.Testing result is as shown in figure 4, as seen from Figure 4, prepared enzyme biologic sensor is to 5
×10-6~3 × 10-3The hydrogen peroxide of M concentration ranges has good linear response, and minimum detection limit reaches 2.94 × 10- 6M。
Claims (10)
1. a kind of method that enzyme biologic sensor is prepared based on microelectrode array, the preparation of photoetching microelectrode array, conductive layer
It prepares and the structure of enzyme biologic sensor is as follows：
(1) preparation of photoetching microelectrode array
One layer of tackifier of spin coating, then coat on negative photoresist after pretreating substrates, carry out front baking, exposure, rear baking, show
Shadow, post bake technique are completed, 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 enzyme biologic sensor
The bioactive layer with specific molecular recognition capability is prepared on the basis of (2), the conduction for first preparing (2) is micro-
Electrod-array, which is dipped in the midbody solution with connection function, to be impregnated, with ultrapure water and with nitrogen (N after taking-up2) blow
It is dry；It is subsequently placed into the enzyme solutions of buffer solution configuration, it is anti-that with enzyme dehydrating condensation can occur for the intermediate that early period is covalently attached
Should, so as to fulfill effective fixation of enzyme, with PBS and ultrapure water and logical nitrogen (N during taking-up2) dry to get to spy
The enzyme biologic sensor of different in nature detection type.
2. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that the base material for silicon chip, glass plate, aluminium flake,
Any one of copper sheet, needs to be pre-processed before use, and processing method is：Substrate surface is cleaned using acetone, is removed
Then the base material cleaned is positioned in the thermal station that temperature is 200 DEG C and toasts 30min to remove base material by surface and oil contaminant and impurity
The solvent on surface.
3. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that the array has the specific surface area of superelevation,
Wherein negative photoresist selection can prepare high aspect ratio micro-nano structure 2000 series 2025 of SU-8 Microchem, 2035,
2050th, any one of 2075, tackifier are OMNICOAT (>=17nm).
4. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that the technological parameter of the photolithographic process is：
(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) it exposes：Wavelength is 365nm, light intensity 12.9mw/cm2, the time is 5s~30s；
(4) it dries 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. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that used in the photolithographic process on mask plate
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, in circular hole
Portion is light transmission part.
6. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that the method for introducing conductive element, except true
Outside these conventional methods of empty evaporation coating, vacuum sputtering coating, vacuum ion membrane plating and chemical vapor deposition, it can also be used indirectly
Method such as forms middle layer using dopamine (DA) adhesiveness, then the method for conductive layer is formed by suction-operated adsorbing metal.
7. the enzyme biologic sensor according to prepared by claim 1, 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 nanotube (CNT), graphene (GN)；The conduction
Layer thickness is 20nm~80nm.
8. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that the intermediate with connection function can
Be in Mercaptoundecanoic acid (MUA), 4- mercaptobenzoic acids (MBA) and glutaraldehyde it is any；A concentration of 5mM~50mM, solvent
For ethanol solution, processing time is 10h~for 24 hours.
9. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that in the enzyme solutions, solvent pH=7.0
Phosphate buffer solution, enzyme is horseradish peroxidase (HRP), lactate oxidase (LOx), glucose oxidase (GOx), courage are solid
Alcoholase, lipase, laccase, tyrosinase, cholinesterase, organophosphor hydrolytic enzyme, nitrate reductase, nitrite reductase, height
One or more in chlorate reductase, urase, carbonic anhydrase, enzyme concentration is 1mg/mL~20mg/mL.
10. the enzyme biologic sensor according to prepared by claim 1, it is characterised in that need to add in the N- of 15mM in the enzyme solutions
The 1- of HOSu NHS (NHS) and 30mM (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC), to
Activated carboxyl accelerates reaction process.
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| CN114894871A (en) * | 2022-05-16 | 2022-08-12 | 安徽大学 | Preparation method and application of high-sensitivity nitrite reductase bioelectrode |
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| CN113549941B (en) * | 2021-07-21 | 2022-08-26 | 东南大学 | Microelectrode array and method for electrochemical synthesis of compound by using same |
| CN113804627A (en) * | 2021-09-14 | 2021-12-17 | 广东中图半导体科技股份有限公司 | Semiconductor photoelectrochemical sensor and preparation method thereof |
| CN114894871A (en) * | 2022-05-16 | 2022-08-12 | 安徽大学 | Preparation method and application of high-sensitivity nitrite reductase bioelectrode |
| CN114894871B (en) * | 2022-05-16 | 2024-01-16 | 安徽大学 | Preparation method and application of high-sensitivity nitrite reductase bioelectrode |
| CN114965643A (en) * | 2022-05-24 | 2022-08-30 | 重庆科技学院 | Cu/Cu 2 O/Ni(OH) 2 Electrode, glucose sensor and application thereof |
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