CN106483118A - A kind of visualization choline sensor based on bipolar electrode array - Google Patents
A kind of visualization choline sensor based on bipolar electrode array Download PDFInfo
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- CN106483118A CN106483118A CN201610820608.8A CN201610820608A CN106483118A CN 106483118 A CN106483118 A CN 106483118A CN 201610820608 A CN201610820608 A CN 201610820608A CN 106483118 A CN106483118 A CN 106483118A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
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Abstract
The invention discloses a kind of visualization choline sensor based on bipolar electrode array, belongs to biosensor technology field.The present invention is based on choline oxidase/Carbon Nanotubes/Chitosan modified electrode, bipolar electrode and microfluidic chip technology, realizes the Visual retrieval of choline using electrochemical luminescence imaging.Due to bipolar electrode be a kind of do not need the external electrical contact can be in the conductive material of its end generation electrochemical reaction, this characteristic makes it easy to carry out a large amount of array analysis, beneficial to production in enormous quantities, so as to reduce testing cost.Using electrochemical luminescence imaging technique have the advantages that sensitivity is high, light path is simple, for the Visual retrieval of choline, method is easy, visual result.
Description
Technical field
The present invention relates to biosensor technology field, more particularly to a kind of visualization choline based on bipolar electrode array
The method of sensor detection choline.
Background technology
Choline is a kind of organic base, is the precursor of acetylcholine and the constituent of lecithin, exists in neurolemma phosphorus
In fat.Choline is adjusting fat metabolism in animal body, the savings of control cholesterol, is preventing fatty liver, guarantee somatic normal
The aspects such as the normal operation of vital movement, promotion cartilage development and nervous system all play an important role, and its detection is with weight
The biological significance that wants.
As choline does not have UV absorption and electro-chemical activity, so becoming the weight for determining choline using the catalytic reaction of enzyme
Want method.Most choline sensors use peroxidase, while by means of electron mediator or nano material.The use of enzyme can
So that the selectivity of sensor is ensured, but the fixing means of enzyme and its activity are still one to be needed to continue asking for research
Topic.Greatly improve can the performance of sensor additionally, introducing medium, but while have also been introduced complexity and limitation.Mesh
But the front biology sensor for choline concentration detection in human serum is it has been reported that as anti-interference is poor, the range of linearity
The weak point such as narrow, service life is short, they are restricted in clinical detection application.
Therefore, developing a kind of new choline sensor that selectivity is good, electrode structure is easy, with low cost will be with important
Research Significance and wide market value.
Content of the invention
In order to make up the deficiencies in the prior art, the invention provides a kind of selectivity is good, electrode structure is easy, with low cost
Choline detection method, the visualization electrochemical luminescence that the method for the present invention is constituted based on bipolar electrode array-micro-fluidic chip
Sensor.
The technical scheme is that：
A kind of preparation method of the visualization choline sensor based on bipolar electrode array, including step：
1) cover plate is prepared
Balance above the PDMS formpiston for being poured on silanization with curing agent after the PDMS mother liquor degassing that PDMS monomer is mixed, then
Preservative film is covered, is heating and curing；After solidification, PDMS is peeled off from the PDMS formpiston of silanization, is obtained containing channel groove
PDMS cover plate；A duct is respectively processed at PDMS cover plate passage two ends, for sample introduction and placement driving electrodes；
2) substrate is prepared
The preparation of A.ITO bipolar electrode substrate
The thick positive optical cement glue-line of 5-15 micron is applied in the conductive surface of ITO electro-conductive glass, by photoetching drawn for laser photocomposing machine
Mask is fitted tightly with the ITO electro-conductive glass for scribbling positive optical cement glue-line, is then carried out mercury lamp exposure, after exposure is put electro-conductive glass
Develop in the NaOH solution of 0.5-1wt% after manifesting to electrode pattern completely and electro-conductive glass is taken out, cleaning, dry；Continuing will
Electro-conductive glass is placed in nitric acid and etching in the mixed aqueous solution of hydrochloric acid, after etching is finished, cleaning, and dry, obtain surface and be carved with three
The substrate of ITO bipolar electrode；
B. choline oxidase/Carbon Nanotubes/Chitosan compound is prepared
Shitosan is dissolved in the aqueous acetic acid of 0.5-2wt%, obtains the chitosan solution of 5-15mg/mL, adds multi-wall carbon nano-tube
Manage and be dispersed in chitosan solution and black suspension is obtained, the addition of CNT is 1-3mg/mL；Concentration is taken for 3-
The choline oxidase solution of 8mg/mL is mixed according to volume ratio 1: 1-3 with the black suspension, obtains choline oxidase/carbon nanometer
The mixed liquor of pipe/Chitosan Composites；
The surface modification of C.ITO bipolar electrode
A rectangle for preventing decorative material solution from spreading is pasted on the anode tap of the ITO bipolar electrode with hydrophobic adhesive tape
Region, pipettes 10-30 microlitre of choline oxidase/Carbon Nanotubes/Chitosan solution and drips in rectangular area surface；Place the substrate in 4
In DEG C refrigerator, dried overnight is to form the ITO bipolar electrode of choline oxidase/Carbon Nanotubes/Chitosan complex thin film modification；
3) chip sealing
Substrate and the cover plate of above-mentioned preparation is cleaned, three ITO bipolar electrodes on substrate is directed at the middle part of cover plate channel grooves, so
After fit tightly, obtain choline electrochemical luminous sensor chip.
Preferably, step 1) in, in PDMS mother liquor, PDMS monomer is 8: 1-12: 1 with the mass ratio of curing agent；
5-20min is placed under 4 DEG C of environment before the degassing of PDMS mother liquor；Degassing time is 20-40min.
Preferably, step 1) in, PDMS mother liquor equilibration time above the PDMS formpiston of silanization is 5-
15min.
Preferably, step 1) in, the duct at PDMS cover plate passage two ends is circular duct.
Preferably, step 2) in, before ITO conductive glass surface applies positive optical cement, ITO electro-conductive glass is carried out
Pretreatment；The pretreatment is specially soaked overnight in the ethanol solution that ITO electro-conductive glass is placed in 0.8-1.2M NaOH, it
It is cleaned by ultrasonic 10-20min in acetone, absolute ethyl alcohol and deionized water respectively afterwards, clean ITO electro-conductive glass is blown with nitrogen
Dry.
Preferably, step 2) in A, uniform drop coating is drawn after positive optical cement in ITO electro-conductive glass with glue head dropper
Conductive surface, using photoresist spinner spin coating under 400-600rpm rotating speed, then rotates 30-50s under 2500-3500rpm rotating speed
Whirl coating；Afterwards ITO electro-conductive glass is placed under 105-115 DEG C of environment and dries, then slowly cool on 25-30 DEG C of hot plate
Room temperature, forms positive optical cement glue-line in uniform thickness.
Preferably, step 2) in A, the time of mercury lamp exposure is 2-6min；Electro-conductive glass is placed in nitric acid and salt
6-15min is etched in the mixed aqueous solution of acid, wherein, in the mixed aqueous solution of nitric acid and hydrochloric acid, the quality of nitric acid, hydrochloric acid and water
Than for 2-3: 4-6: 4-6.
The method for choline being detected using the visualization choline sensor based on bipolar electrode array, at cover plate two ends
A platinum filament is placed on duct respectively, as driving electrodes；Driving electrodes provide driving voltage by D.C. regulated power supply；Xiang Tong
Cushioning liquid of the injection containing choline solution to be measured and luminol in road groove；The choline oxidase of bipolar electrode surface modification is urged
Change dissolved oxygen H is generated with acetylcholine response2O2, and under alive driving outside, there is electrochemical luminescence with luminol, generation
Electrochemical luminescence signals are by the ccd image sensor capture above choline electrochemical luminous sensor chip；According to ccd image
The luminous signal intensity of sensor capture determines the choline concentration in choline solution to be measured.
Preferably, the 15-30 microlitre of buffering containing choline solution to be measured and luminol is injected into channel groove
Solution；The cushioning liquid is PBS cushioning liquid；The concentration of PBS in cushioning liquid containing choline solution to be measured and luminol
It is 7.2-7.5 for 0.05-0.15mol/L, pH；The concentration of choline is 0.01mM-1mM；The concentration of luminol is 1.2-
1.4mmol/L.Present invention detection liquor capacity used is only 15-30 microlitre, can achieve trace detection, greatly improves detection
Sensitivity, and reduce testing cost.
Preferably, the driving voltage is 4.0-9.0V.The possible ito film of overtension can be destroyed, therefore electricity
Pressure selects the scope.
Idea of the invention is that：
With the complex film modified ITO bipolar electrode of choline oxidase/Carbon Nanotubes/Chitosan as sensing interface, by micro-fluidic
Chip system, platinum filament driving electrodes, D.C. regulated power supply and ccd image sensor realize the visualization electrochemical luminescence inspection of choline
Survey.Driving electrodes provide driving voltage by D.C. regulated power supply.Into choline electrochemical luminous sensor chip channel groove
Choline solution to be measured and the cushioning liquid containing luminol is injected, the choline oxidase of bipolar electrode surface modification can be catalyzed dissolving
Oxygen generates H with acetylcholine response2O2, and under alive driving outside, there is electrochemical luminescence with luminol.The electrochemistry of generation
Luminous signal is captured by the ccd image sensor above sensing chip.Within the specific limits, choline concentration is higher, enzymatic reaction
The hydrogen peroxide of generation is more, and corresponding luminous signal is also stronger, it is possible thereby to realize the Visual retrieval of choline.
Present invention visualization electrochemical luminous sensor detects that the measuring principle of choline is：
Catalytic reaction on bipolar electrode anode tap modified membrane：
Choline generates trimethylglycine and hydrogen peroxide with oxygen in the presence of choline oxidase；
The electrochemical reaction of bipolar electrode anode tap：
Luminol+H2O2→3-Aminophthalate+hv
Luminol generates 3- amino-repefral and with luminous with hydrogen peroxide reaction；
The electrochemical reaction of bipolar electrode cathode terminal：
4H2O+4e-→2H2+4OH-
The present invention is based on choline oxidase/Carbon Nanotubes/Chitosan modified electrode, bipolar electrode and microfluidic chip technology, profit
Electricity consumption chemiluminescence imaging is realizing the Visual retrieval of choline.As bipolar electrode is that one kind does not need external electrical contact
There is the conductive material of electrochemical reaction in its end, this characteristic makes it easy to a large amount of array analysis are carried out, beneficial to high-volume
Production, so as to reduce testing cost.
Using electrochemical luminescence imaging technique have the advantages that sensitivity is high, light path is simple, for the visualization of choline
Detection, method is easy, visual result.
Electrode modified material choline oxidase/Carbon Nanotubes/Chitosan in the present invention, choline oxidase choline oxidase,
CNT, shitosan are indispensable, have choline oxidase just have catalytic reaction to produce hydrogen peroxide, and then send out with luminol
Hair tonic light reaction；CNT is to promote electrochemical reaction, is obstructed without CNT electrochemical reaction, so also not having
Light is produced；Shitosan is filmogen, if without shitosan, choline oxidase, carbon nano-tube solution stably cannot be attached to
Electrode surface.
Beneficial effects of the present invention are：
Preparation method of the present invention is simple and convenient, and choline oxidase/Carbon Nanotubes/Chitosan composite membrane fixes can choline oxidase
In electrode surface, there are preferable film forming and biocompatibility；CNT promotes Electron Transfer, and shitosan promotes film forming；
The sensor has that specific good, sensitivity is high, selective advantage, and material used is cheap and easy to get.
Open bipolar electrode due to without external electrical contact, in a microchannel, luminescence reagent and detected material
Can be blended in a solution, large scale array analysis can be carried out, fast high-flux detects choline.
Microfluidic chip structure is simple, prepares simple, with low cost, integrated micro, beneficial to production in enormous quantities, substantially reduces
Testing cost, is expected to be applied to the exploitation of choline test paper.
The high sensitivity of electrochemical luminescence imaging technique, time and space are differentiated, the optics of simplification, and visual imaging energy
Power is visualized, and is to detect that the content of choline provides a sensitive, easy and intuitively Sparklet testing method.
The preparation method can be generalized to other types enzyme, for the detection of other biochemical moleculars, can develop multicomponent
The bipolar electrode sensor of detection.
Description of the drawings
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
Accompanying drawing to be used needed for technology description is had to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, on the premise of not paying creative work, acceptable
Other accompanying drawings are obtained according to these accompanying drawings.
Fig. 1 is device and reaction principle schematic diagram of the present invention based on the visualization choline sensor of bipolar electrode array；
Fig. 2 is the structural representation of ITO bipolar electrode array of the present invention；
Fig. 3 is the machining sketch chart of ITO bipolar electrode array chip of the present invention
Fig. 4 is the electrochemical luminescence figure of choline of the present invention detection；Wherein, a corresponding concentration 0mM；B corresponding concentration 0.02mM；C is corresponded to
Concentration 0.05mM；D corresponding concentration 0.1mM；E corresponding concentration 0.5mM；F corresponding concentration 1.0mM；G corresponding concentration 2.0mM；H is corresponded to
Concentration 5.0mM.
Fig. 5 is the linear graph of choline of the present invention detection.
Specific embodiment
Preparation of the embodiment 1 based on the visualization choline sensor of bipolar electrode array
As shown in Figure 1, Figure 2, Figure 3 shows, a kind of preparation method of the visualization choline sensor based on bipolar electrode array, including step
Suddenly：
1) cover plate is prepared
By PDMS monomer and curing agent with 8: 1-12: 1 mass ratio is mixed and stirred for uniform PDMS mother liquor, by PDMS mother liquor in
5-20min is placed under 4 DEG C of environment, and then deaerate 2040min；Balance 5- above the PDMS formpiston of silanization is poured on after degassing
15min, then covers preservative film, the 70-80 DEG C of 1-2 hour that is heating and curing；After solidification, by PDMS from the PDMS formpiston of silanization
Peel off, the cover plate of 3.0cm*1.2cm is cut into, on the cover plate, contains channel groove；Flat mouth syringe needle with 14G is in PDMS cover plate passage
Two ends respectively process the circular duct of an a diameter of 0.5cm, for sample introduction and placement driving electrodes；
2) substrate is prepared
A. there is the preparation of three ITO bipolar electrode substrates
ITO electro-conductive glass is cut into the small pieces of 3.0cm*1.2cm；The ITO electro-conductive glass of well cutting is placed in 0.8-1.2M
Soaked overnight in the ethanol solution of NaOH, is cleaned by ultrasonic 10-20min afterwards respectively in acetone, absolute ethyl alcohol and deionized water,
Clean ITO electro-conductive glass is dried up with nitrogen；Conducting surface is determined with universal meter, conduction faces up and is placed on photoresist spinner disk；
Uniform drop coating is drawn after the positive optical cement of AZP4620 in the conductive surface of ITO electro-conductive glass with glue head dropper, in 400-600rpm rotating speed
Lower spin coating, then rotates 30-50s whirl coating under 2500-3500rpm rotating speed；The conductive surface for staying in ITO electro-conductive glass applies 5-15
After the thick red light glue-line of micron, ITO electro-conductive glass is placed under 105-115 DEG C of environment and is dried, then in 25-30 DEG C of hot plate
On be slowly cooled to room temperature, form positive optical cement glue-line in uniform thickness.
Photo etched mask drawn for laser photocomposing machine is fitted tightly with the ITO electro-conductive glass for scribbling positive optical cement glue-line, so
After carry out selective mercury lamp exposure 2-6min, develop to electricity in the NaOH solution that electro-conductive glass is placed in after exposure 0.5-1wt%
Pole figure case takes out electro-conductive glass after manifesting completely；Cleaning removes remaining developer solution in deionized water, is subsequently blown with hair-dryer
Dry electro-conductive glass, then dries 30min under 110 DEG C of hot plates, and baking terminates for hot plate temperature to be adjusted to 20-25 DEG C, delays on hot plate
Room temperature is down to slowly.
Continue by electro-conductive glass be placed in nitric acid and hydrochloric acid mixed aqueous solution (mass ratio of nitric acid, hydrochloric acid and water is 2-3:
Etching 6-15min in 4-6: 4-6), after etching is finished, deionized water, acetone, deionized water are cleaned by ultrasonic each 10min successively,
Dry, obtain the substrate that three ITO bipolar electrodes are carved with surface；
B. choline oxidase/Carbon Nanotubes/Chitosan compound is prepared
Shitosan is dissolved in the aqueous acetic acid of 0.5-2wt%, ultrasonic dissolution, obtains the chitosan solution of 5-15mg/mL, is added many
Wall carbon nano tube ultrasonic disperse in the chitosan solution black suspension, the addition of CNT is 1-3mg/mL；Take
Concentration is mixed according to volume ratio 1: 1-3 with the black suspension for the choline oxidase solution of 3-8mg/mL, and ultrasonic dissolution is obtained
The mixed liquor of choline oxidase/Carbon Nanotubes/Chitosan Composites, 4 DEG C of preservations；
The surface modification of C.ITO bipolar electrode
A rectangle for preventing decorative material solution from spreading is pasted on the anode tap of the ITO bipolar electrode with hydrophobic adhesive tape
Region, pipettes 10-30 microlitre of choline oxidase/Carbon Nanotubes/Chitosan solution and drips in rectangular area surface；Chip is placed in 4
In DEG C refrigerator, dried overnight is to form the ITO bipolar electrode of choline oxidase/Carbon Nanotubes/Chitosan complex thin film modification；
3) chip sealing
The cover plate that has modified and substrate are respectively placed in plasma clean room, each process 90 seconds and 25 seconds.By on substrate three
The middle part of root ITO bipolar electrode alignment cover plate channel groove, then fits tightly, obtains choline electrochemical luminous sensor chip.
Embodiment 2 is used for the detection of choline based on the visualization choline sensor of bipolar electrode array
Method using choline is detected based on the visualization choline sensor of bipolar electrode array, divides on the duct at cover plate two ends
Not Fang Zhi a platinum filament, as driving electrodes；Driving electrodes provide the driving voltage of 8V by D.C. regulated power supply；Recessed to passage
Cushioning liquid of 20 microlitres of the injection containing choline solution to be measured and luminol in groove, wherein, the concentration of PBS is 0.01mol/L, pH
For 7.4, the concentration of choline is 0.02mM-5mM, and the concentration of luminol is 1.3mmol/L；The choline oxygen of bipolar electrode surface modification
Change enzymatic dissolved oxygen H is generated with acetylcholine response2O2, and under alive driving outside, there is electrochemical luminescence with luminol,
The electrochemical luminescence signals of generation are by the ccd image sensor capture above choline electrochemical luminous sensor chip；According to
The luminous signal intensity of ccd image sensor capture determines the choline concentration in choline solution to be measured.
Choline to 0mM, 0.02mM, 0.05mM, 0.1mM, 0.5mM, 1.0mM, 2.0mM and 5.0mM series concentration respectively
Detected, testing result is as shown in Figure 4 and Figure 5；The minimum detectability of the inventive method detection choline is 43.19 μM；With
The rising of choline concentration, the gray value of luminescent image are also gradually increased.By linear fit, gray value is existed with choline concentration
There is good linear relationship in the range of 0.02-5.0mM.Gained fit equation is：
G=1022+212.2 [choline] (mM) (R=0.9924).
The preferred embodiments of the present invention are the foregoing is only, the present invention is not limited to, although with reference to aforementioned reality
Apply example the present invention has been described in detail, those skilled in the art are said, which still can be to aforementioned each enforcement
Technical scheme described in example is modified, or carries out equivalent to which part technical characteristic.All essences in the present invention
Within god and principle, any modification for being made, equivalent.Improve etc., should be included within the scope of the present invention.
Claims (10)
1. a kind of based on bipolar electrode array visualization choline sensor preparation method, it is characterised in that including step：
1) cover plate is prepared
Balance above the PDMS formpiston for being poured on silanization with curing agent after the PDMS mother liquor degassing that PDMS monomer is mixed, then
Preservative film is covered, is heating and curing；After solidification, PDMS is peeled off from the PDMS formpiston of silanization, is obtained containing channel groove
PDMS cover plate；A duct is respectively processed at PDMS cover plate passage two ends, for sample introduction and placement driving electrodes；
2) substrate is prepared
The preparation of A.ITO bipolar electrode substrate
The thick positive optical cement glue-line of 5-15 micron is applied in the conductive surface of ITO electro-conductive glass, by photoetching drawn for laser photocomposing machine
Mask is fitted tightly with the ITO electro-conductive glass for scribbling positive optical cement glue-line, is then carried out mercury lamp exposure, after exposure is put electro-conductive glass
Develop in the NaOH solution of 0.5-1wt% after manifesting to electrode pattern completely and electro-conductive glass is taken out, cleaning, dry；Continuing will
Electro-conductive glass is placed in nitric acid and etching in the mixed aqueous solution of hydrochloric acid, after etching is finished, cleaning, and dry, obtain surface and be carved with three
The substrate of ITO bipolar electrode array；
B. choline oxidase/Carbon Nanotubes/Chitosan compound is prepared
Shitosan is dissolved in the aqueous acetic acid of 0.5-2wt%, obtains the chitosan solution of 5-15mg/mL, adds multi-wall carbon nano-tube
Manage and be dispersed in chitosan solution and black suspension is obtained, the addition of CNT is 1-3mg/mL；Concentration is taken for 3-
The choline oxidase solution of 8mg/mL is mixed according to volume ratio 1: 1-3 with the black suspension, obtains choline oxidase/carbon nanometer
The mixed liquor of pipe/Chitosan Composites；
The surface modification of C.ITO bipolar electrode
A rectangle for preventing decorative material solution from spreading is pasted on the anode tap of the ITO bipolar electrode with hydrophobic adhesive tape
Region, pipettes 10-30 microlitre of choline oxidase/Carbon Nanotubes/Chitosan solution and drips in rectangular area surface；Place the substrate in 4
In DEG C refrigerator, dried overnight is to form the ITO bipolar electrode of choline oxidase/Carbon Nanotubes/Chitosan complex thin film modification；
3) chip sealing
Substrate and the cover plate of above-mentioned preparation is cleaned with plasma cleaning device, and three ITO bipolar electrode alignment cover plates on substrate are led to
The middle part of road groove, then fits tightly, and obtains choline electrochemical luminous sensor chip.
2. preparation method as claimed in claim 1 based on the visualization choline sensor of bipolar electrode array, it is characterised in that：
Step 1) in, in PDMS mother liquor, PDMS monomer is 8: 1-12: 1 with the mass ratio of curing agent；In 4 DEG C of environment before the degassing of PDMS mother liquor
Lower placement 5-20min；Degassing time is 20-40min.
3. preparation method as claimed in claim 1 or 2 based on the visualization choline sensor of bipolar electrode array, its feature exist
In：Step 1) in, PDMS mother liquor equilibration time above the PDMS formpiston of silanization is 5-15min.
4. preparation method as claimed in claim 1 or 2 based on the visualization choline sensor of bipolar electrode array, its feature exist
In：Step 1) in, the duct at PDMS cover plate passage two ends is circular duct.
5. preparation method as claimed in claim 1 based on the visualization choline sensor of bipolar electrode array, it is characterised in that：
Step 2) in, before ITO conductive glass surface applies positive optical cement, ITO electro-conductive glass is pre-processed；The pretreatment is concrete
It is soaked overnight in the ethanol solution for be placed in ITO electro-conductive glass 0.8-1.2M NaOH, afterwards respectively in acetone, absolute ethyl alcohol
It is cleaned by ultrasonic 10-20min with deionized water, clean ITO electro-conductive glass is dried up with nitrogen.
6. as described in claim 1 or 5 based on bipolar electrode array visualization choline sensor preparation method, its feature exists
In：Step 2) in A, uniform drop coating is drawn after positive optical cement in the conductive surface of ITO electro-conductive glass with glue head dropper, using photoresist spinner
Spin coating under 400-600rpm rotating speed, then rotates 30-50s whirl coating under 2500-3500rpm rotating speed；Afterwards by ITO conduction glass
Glass is placed under 105-115 DEG C of environment and dries, and is then slowly cooled to room temperature on 25-30 DEG C of hot plate, is formed in uniform thickness
Positive optical cement glue-line.
7. as described in claim 1 or 5 based on bipolar electrode array visualization choline sensor preparation method, its feature exists
In：Step 2) in A, the time of mercury lamp exposure is 2-6min；Electro-conductive glass is placed in mixed aqueous solution of the nitric acid with hydrochloric acid and is carved
Erosion 6-15min, wherein, in the mixed aqueous solution of nitric acid and hydrochloric acid, the mass ratio of nitric acid, hydrochloric acid and water is 2-3: 4-6: 4-6.
8. using the method for detecting choline based on the visualization choline sensor of bipolar electrode array as claimed in claim 1, its
It is characterised by：A platinum filament is placed on the duct at cover plate two ends respectively, as driving electrodes；Driving electrodes pass through DC voltage-stabilizing
Power supply provides driving voltage；Cushioning liquid of the injection containing choline solution to be measured and luminol into channel groove；Bipolar electrode
The choline oxidase catalysis dissolved oxygen of surface modification generates H with acetylcholine response2O2, and under alive driving outside, with luminol
Generation electrochemical luminescence, the electrochemical luminescence signals of generation are by the ccd image above choline electrochemical luminous sensor chip
Sensor is captured；Luminous signal intensity according to ccd image sensor capture determines the choline concentration in choline solution to be measured.
9. the method for as claimed in claim 8 choline being detected based on the visualization choline sensor of bipolar electrode array, its feature
It is：The 15-30 microlitre of cushioning liquid containing choline solution to be measured and luminol is injected into channel groove；The cushioning liquid
For PBS cushioning liquid；In cushioning liquid containing choline solution to be measured and luminol, the concentration of PBS is 0.05-0.15mol/L,
PH is 7.2-7.5；The concentration of choline is 0.02mM-5mM；The concentration of luminol is 1.2-1.4mmol/L.
10. the method for choline being detected based on the visualization choline sensor of bipolar electrode array as described in claim 8 or 9, its
It is characterised by：The driving voltage is 4.0-9.0V.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN112362708A (en) * | 2020-10-19 | 2021-02-12 | 济南大学 | Preparation method of self-powered bipolar microelectrode microfluidic chip photoelectrochemical aptamer sensor |
| CN114062470A (en) * | 2021-09-29 | 2022-02-18 | 福建医科大学 | 3D printing array type BP-LED-E sensing device for hydrogen peroxide detection |
| CN114062470B (en) * | 2021-09-29 | 2023-06-13 | 福建医科大学 | 3D printing array type BP-LED-E sensing device for hydrogen peroxide detection |
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