CN103558266A - Graphene capacitive biosensor and methods for manufacturing and detecting same - Google Patents
Graphene capacitive biosensor and methods for manufacturing and detecting same Download PDFInfo
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- CN103558266A CN103558266A CN201310508400.9A CN201310508400A CN103558266A CN 103558266 A CN103558266 A CN 103558266A CN 201310508400 A CN201310508400 A CN 201310508400A CN 103558266 A CN103558266 A CN 103558266A
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Abstract
The invention discloses a graphene capacitive biosensor and methods for manufacturing and detecting the same. The graphene capacitive biosensor comprises a glass substrate, wherein indium tin oxides (ITO) are arranged on the two sides of the glass substrate respectively; graphene covers part of the glass substrate on the same side and the ITOs; the glass substrate which is not covered with the graphene and the ITOs on the two sides are positioned on the same side; each of the ITOs on the two sides of the glass substrate which is not covered with the graphene is an electrode of a capacitor; polyethylene terephthalate (PET) gaskets cover the ITOs which are covered with the graphene; a PET substrate to which a gold film is sputtered covers each PET gasket; a sample cell is formed in the middle of the glass substrate by insulating silicon gel; the gold film is the other electrode of the capacitor. By using a method for covering the electrodes of the ITOs with the graphene, the contact resistance is reduced, instantaneous voltage pulse is applied, and the capacitor responds for measurement. Adenosine triphosphate is taken as an experiment object, and the detection lower limit of the adenosine triphosphate reaches 1pM through the method.
Description
Technical field
The present invention relates to analytical chemistry field, relate in particular to a kind of Graphene capacitor biological sensor and preparation method thereof, detection method.
Background technology
Graphene is a kind of emerging functional material.Due to its characteristic that there is good optics, electricity and be easy to be combined by the effect of π-π stacking with biomolecule, and receive publicity in biosensor design field.Biomolecule is adsorbed on the channel doping that Graphene surface can regulate its electric charge transmission or electric charge, causes the variation of carrier concentration and carrier mobility.
Graphene ultracapacitor is the general designation of the ultracapacitor based on grapheme material.Due to the two-dimensional structure of Graphene uniqueness and outstanding intrinsic physical characteristics, such as abnormal high electric conductivity and high surface area, the application of graphene-based material in ultracapacitor has great potentiality.Graphene-based material is compared with traditional electrode material, in the process of energy storage and release, has shown the Characteristics and mechanism that some are novel.
Graphene biosensor based on capacity effect there is not yet report.The theory structure formula of its principle based on capacitor:
C=εS/4πkd.
Wherein, the capacity that C is capacitor, S is capacitor plate area, and d is capacitor plate distance, and ε is dielectric specific inductive capacity between plate.When dielectric concentration change, ε changes the variation that causes condenser capacitance.Therefore, change dielectric concentration, can realize the detection to dielectric concentration by measuring condenser capacitance.
Summary of the invention
The invention discloses a kind of Graphene capacitor biological sensor and preparation method thereof, detection method, the method that adopts Graphene to cover ITO electrode reduces contact resistance, as a utmost point of capacitance electrode, adopts plane gold electrode as another utmost point of capacitance electrode.
For achieving the above object, concrete scheme of the present invention is as follows:
A kind of Graphene capacitor biological sensor, comprise substrate of glass, the both sides of substrate of glass are equipped with ITO, above the substrate of glass of part homonymy and ITO, be coated with Graphene, do not cover the substrate of glass of Graphene and the ITO of both sides is positioned at homonymy, the utmost point that the ITO that does not cover the both sides of Graphene is electric capacity, be coated with Graphene ITO be coated with above PET pad, on PET pad, be coated with the PET substrate of sputter gold film, utilize insulation silica gel at the middle part of substrate of glass, to be provided with a sample cell, another utmost point that golden film is electric capacity.
Describedly do not cover the substrate of glass of Graphene and the ITO size of both sides is respectively 20mm*20mm and 20mm*5mm.
Described sample cell is of a size of: 20mm*10mm*3mm.
The thickness of described ITO is 200nm.
Described PET shim size is 20*5mm, and thickness is 3mm.
Described PET substrate is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film is 150nm.
A method for making for Graphene capacitor biological sensor, comprises the following steps:
Step 1: utilize chemical gaseous phase depositing process growth single or multiple lift Graphene and its transfer is covered to segment glass substrate, tin indium oxide is located on the both sides of substrate of glass and is positioned under Graphene and with Graphene and contacts without the residual one side of PMMA;
Step 2: Graphene is covered to after substrate of glass, with insulation silica gel, surround sample cell, utilize magnetically controlled sputter method, sputter gold film in PET substrate, and another using golden film as electric capacity be fixed on sample cell with insulation silica gel extremely downwards, thereby then application of sample forms integrated single graphene device.
In described step 1, substrate of glass is of a size of 20*20mm, and indium-tin oxide electrode is of a size of 20*5mm, and thickness is 200nm; The transfer method that utilizes chemical gaseous phase depositing process growth single or multiple lift Graphene in described step 1 and its transfer is covered to segment glass substrate is wet method transfer method;
The top of tin indium oxide in described step 1, utilizing insulation silica gel fixed measure is 20*5mm, the PET pad that thickness is 3mm;
In described step 1, on not capped tin indium oxide, smear conductive silver paste extraction electrode respectively as the utmost point of electric capacity;
In described step 1, a utmost point of the electric capacity of Graphene and another electrode resistance of electric capacity are 1K ohm;
In described step 2, PET substrate is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film is 150nm;
In described step 2, sputter is had to the PET substrate of golden film, golden film utilizes insulation silica gel to be covered on PET pad downwards, thereby forms the sample cell that is of a size of 20mm*10mm*3mm;
A detection method for Graphene capacitor biological sensor, comprises the following steps:
Step 1: by Graphene capacitor biological sensor detection circuit for access;
Step 2: use pipettor to add 300 μ L ATP samples in sample cell, processor changes D/A voltage to be exported to 2.5V to 1.5V by 0V to 1.5V, the voltage Dynamic Signal of Detection capacitance device, in real time calculable capacitor capacitance;
In described step 1, testing circuit is: a utmost point ground connection of electric capacity, another utmost point of electric capacity is connected with variable voltage source by R1, variable voltage source is connected with the D/A end of microprocessor, R2 is in parallel with electric capacity and the common port of R2, R1 and electric capacity and one end input end of instrument amplifier are connected, the common port on R2, electric capacity and ground is connected with the other end input end of instrument amplifier by resistance R, the output terminal of instrument amplifier is connected with the A/D of microprocessor end, and microprocessor is connected with computing machine.
Described resistance R
1=R
2=1K ohm;
The scope that described microprocessor gathers voltage is 0-2.5V, and A/D conversion accuracy is minimum is 12, and sample rate is at least 1Kbit/s;
Described D/A conversion unit applies voltage to another utmost point of electric capacity, and voltage conversion range is 0-2.5V, and conversion accuracy is 10;
Described capacitor electrode capacitance computation process: select time constant t=RC, capacitance voltage is charged to 0.63 times of D/A output voltage, and the performance graph constant computing time t according to measuring, obtains capacitor electrode capacitance according to formula C=t/R.
Microprocessor loads on testing circuit by controlling D/A switch unit generation voltage signal; Testing circuit discharges and recharges Graphene electric capacity by resistance R.Because capacitance size is relevant with biomolecule concentration in electrolyte, so the time that discharges and recharges is relevant with object concentration to be measured.Based on this principle, capacitor charge and discharge dynamic process is measured, realize the detection to target molecule.
Beneficial effect of the present invention:
(1) method that adopts Graphene to cover ITO electrode reduces contact resistance, applies the response of transient voltage pulsed capacitance and measures.Take atriphos as experimental subjects, and the lower limit of realizing detection atriphos by said method reaches 1pM, and performance is except high sensitivity.
(2) by changing voltage swing and the polarity at electric capacity the two poles of the earth, realize the detection of dynamic to electric capacity, thereby realized the detection of dynamic to dielectric and Graphene interaction process, as shown in Figure 3.
Accompanying drawing explanation
Fig. 1, the making schematic diagram of Graphene capacitor biological sensor;
Fig. 2, the testing circuit schematic diagram of Graphene capacitor biological sensor;
Fig. 3, test pattern one;
Fig. 4, test pattern two;
Wherein, 1 substrate of glass, 2ITO, 3 Graphenes, 4PET pad, 5PET substrate, 6 gold medal films, a utmost point of 7 electric capacity, another utmost point of 8 electric capacity.
Embodiment:
Below in conjunction with accompanying drawing, the present invention is described in detail:
As shown in Figure 1, a kind of Graphene capacitor biological sensor, comprise substrate of glass 1, the both sides of substrate of glass 1 are equipped with ITO2, above the substrate of glass 1 of part homonymy and ITO2, be coated with Graphene 3, do not cover the substrate of glass 1 of Graphene 3 and the ITO2 of both sides and be positioned at homonymy, the utmost point 7 that the ITO2 that does not cover the both sides of Graphene 3 is electric capacity, be coated with Graphene 3 ITO2 be coated with above PET pad 4, on PET pad 4, be coated with the PET substrate 5 of sputter gold film 6, utilize insulation silica gel to be provided with a sample cell at the middle part of substrate of glass 1, another utmost point 8 that gold film 6 is electric capacity.
Describedly do not cover the substrate of glass 1 of Graphene 3 and the ITO2 size of both sides is respectively 20mm*20mm and 20mm*5mm.
Described sample cell is of a size of: 20mm*10mm*3mm.
The thickness of described ITO2 is 200nm.
Described PET pad 4 is of a size of 20*5mm, and thickness is 3mm.
Described PET substrate 5 is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film 6 is 150nm.
A method for making for Graphene capacitor biological sensor, comprises the following steps:
Step 1: utilize chemical gaseous phase depositing process growth single or multiple lift Graphene 3 and its transfer is covered to segment glass substrate 1, tin indium oxide 2 is located on the both sides of substrate of glass 1 and is positioned under Graphene 3 and with Graphene 3 and contacts without the residual one side of PMMA;
Step 2: Graphene 3 is covered to after substrate of glass 1, with insulation silica gel, surround sample cell, utilize magnetically controlled sputter method, sputter gold film 6 in PET substrate 5, and another utmost point 8 using golden film 6 as electric capacity is fixed on sample cell with insulation silica gel downwards, thereby then application of sample forms integrated single graphene device.
In described step 1, substrate of glass 1 is of a size of 20*20mm, and tin indium oxide 2 electrode sizes are 20*5mm, and thickness is 200nm; The transfer method that utilizes chemical gaseous phase depositing process growth single or multiple lift Graphene 3 in described step 1 and its transfer is covered to segment glass substrate 1 is wet method transfer method;
The top of tin indium oxide 2 in described step 1, utilizing insulation silica gel fixed measure is 20*5mm, the PET pad 4 that thickness is 2mm;
In described step 1, on not capped tin indium oxide 2, smear conductive silver paste extraction electrode respectively as the utmost point 7 of electric capacity;
In described step 1,8 resistance of another utmost point of a utmost point 7 of the electric capacity of Graphene 3 and electric capacity are 1K ohm;
In described step 2, PET substrate 5 is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film 6 is 150nm;
In described step 2, sputter is had to the PET substrate 5 of golden film 6, golden film 6 utilizes insulation silica gel to be covered on PET pad 4 downwards, thereby forms the sample cell that is of a size of 20mm*10mm*3mm;
A detection method for Graphene capacitor biological sensor, comprises the following steps:
Step 1: by Graphene capacitor biological sensor detection circuit for access;
Step 2: use pipettor to add 300 μ L ATP samples in sample cell, processor changes D/A voltage to be exported to 2.5V to 1.5V by 0V to 1.5V, the voltage change signal of Real-time and Dynamic Detection capacitor, and real-time calculable capacitor capacitance;
In described step 1, testing circuit is: a utmost point ground connection of electric capacity, another utmost point of electric capacity is connected with variable voltage source by R1, variable voltage source is connected with the D/A end of microprocessor, R2 is in parallel with electric capacity and the common port of R2, R1 and electric capacity and one end input end of instrument amplifier are connected, the common port on R2, electric capacity and ground is connected with the other end input end of instrument amplifier by resistance R, the output terminal of instrument amplifier is connected with the A/D of microprocessor end, and microprocessor is connected with computing machine.
Described resistance R
1=R
2=1K ohm;
The scope that described microprocessor gathers voltage is 0-2.5V, and A/D conversion accuracy is minimum is 12, and sample rate is at least 1Kbit/s;
Described D/A conversion unit applies voltage to another utmost point of electric capacity, and voltage conversion range is 0-2.5V, and conversion accuracy is 10;
Described capacitor electrode capacitance computation process: select time constant t=RC, capacitance voltage is charged to 0.63 times of D/A output voltage, and the performance graph constant computing time t according to measuring, obtains capacitor electrode capacitance according to formula C=t/R.
Graphene FET biology sensor detection method as shown in Figure 2.
(1) CPU changes output variable voltage signal by D/A, after variable voltage source conversion, is loaded in metering circuit, and D/A conversion accuracy is 10.Wherein, voltage-regulation scope is 0-2.5V, resistance R 1=R2=1K ohm.
(2) adopt instrument amplifier to measure the above earth potential of electric capacity, the gain amplifier of instrument amplifier is adjustable.After analog to digital conversion, by microprocessor, gathered, the scope that microprocessor gathers voltage is 0-2.5V.A/D conversion accuracy is minimum is 12, and sample rate is at least 1Kbit/s.
(4) by USB or serial ports, the voltage value of collection is uploaded to computing machine.
Testing result as shown in Figure 3-4, as can be seen from Figure 3, when changing between pole plate voltage, effect due to capacitor, detectable voltage signals is dynamic change, reflected ATP molecule in dielectric and dynamic interaction process between Graphene, the interaction that has disclosed both changes by exponential damping law.Fig. 3 can find out simultaneously, and capacitor dynamic response shows has high sensitivity to ATP concentration.Magnitude of voltage calculable capacitor electric capacity when choosing capacitance voltage and being 1.5V, with ATP concentration relationship as shown in Figure 4.As seen from the figure, under logarithmic coordinate, the capacitance of Graphene capacitor and ATP concentration are good linear relationship, detect ATP concentration limit and can reach 1pM, cash out high sensitivity.
Claims (10)
1. a Graphene capacitor biological sensor, it is characterized in that, comprise substrate of glass, the both sides of substrate of glass are equipped with ITO, above the substrate of glass of part homonymy and ITO, be coated with Graphene, do not cover the substrate of glass of Graphene and the ITO of both sides is positioned at homonymy, the utmost point that the ITO that does not cover the both sides of Graphene is electric capacity, be coated with Graphene ITO be coated with above PET pad, on PET pad, be coated with the PET substrate of sputter gold film, utilize insulation silica gel at the middle part of substrate of glass, to be provided with a sample cell, another utmost point that golden film is electric capacity.
2. a kind of Graphene capacitor biological sensor as claimed in claim 1, is characterized in that, does not describedly cover the substrate of glass of Graphene and the ITO size of both sides is respectively 20mm*20mm and 20mm*5mm;
Described sample cell is of a size of: 20mm*10mm*3mm;
The thickness of described ITO is 200nm.
3. a kind of Graphene capacitor biological sensor as claimed in claim 1, is characterized in that, described PET shim size is 20*5mm, and thickness is 3mm;
Described PET substrate is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film is 150nm.
4. the method for making of a kind of Graphene capacitor biological sensor as claimed in claim 1, is characterized in that, comprises the following steps:
Step 1: utilize chemical gaseous phase depositing process growth single or multiple lift Graphene and its transfer is covered to segment glass substrate, tin indium oxide is located on the both sides of substrate of glass and is positioned under Graphene and with Graphene and contacts without the residual one side of PMMA;
Step 2: Graphene is covered to after substrate of glass, with insulation silica gel, surround sample cell, utilize magnetically controlled sputter method, sputter gold film in PET substrate, and another using golden film as electric capacity be fixed on sample cell with insulation silica gel extremely downwards, thereby then application of sample forms integrated single graphene device.
5. the method for making of a kind of Graphene capacitor biological sensor as claimed in claim 4, is characterized in that, in described step 1, substrate of glass is of a size of 20*20mm, and indium-tin oxide electrode is of a size of 20*5mm, and thickness is 200nm; The transfer method that utilizes chemical gaseous phase depositing process growth single or multiple lift Graphene in described step 1 and its transfer is covered to segment glass substrate is wet method transfer method;
The top of tin indium oxide in described step 1, utilizing insulation silica gel fixed measure is 20*5mm, the PET pad that thickness is 3mm;
In described step 1, on not capped tin indium oxide, smear conductive silver paste extraction electrode respectively as the utmost point of electric capacity;
In described step 1, a utmost point of the electric capacity of Graphene and another electrode resistance of electric capacity are 1K ohm.
6. the method for making of a kind of Graphene capacitor biological sensor as claimed in claim 4, is characterized in that, in described step 2, PET substrate is of a size of 20*20mm, and thickness is 2mm, and the thickness of golden film is 150nm;
In described step 2, sputter is had to the PET substrate of golden film, golden film utilizes insulation silica gel to be covered on PET pad downwards, thereby forms the sample cell that is of a size of 20mm*10mm*3mm.
7. the detection method of a kind of Graphene capacitor biological sensor as claimed in claim 1, is characterized in that, comprises the following steps:
Step 1: by Graphene capacitor biological sensor detection circuit for access;
Step 2: use pipettor to add 300 μ L ATP samples in sample cell, processor changes D/A voltage and press 0V to 1.5V to the output of 2.5V to 1.5V rule, the voltage change process of real-time Detection capacitance device, and calculable capacitor capacitance thus.
8. the detection method of a kind of Graphene capacitor biological sensor as claimed in claim 7, it is characterized in that, in described step 1, testing circuit is: a utmost point ground connection of electric capacity, another utmost point of electric capacity is connected with variable voltage source by R1, variable voltage source is connected with the D/A end of microprocessor, R2 and electric capacity are in parallel and R2, the common port of R1 and electric capacity is connected with one end input end of instrument amplifier, R2, the common port on electric capacity and ground is connected with the other end input end of instrument amplifier by resistance R, the output terminal of instrument amplifier is connected with the A/D of microprocessor end, microprocessor is connected with computing machine.
9. the detection method of a kind of Graphene capacitor biological sensor as claimed in claim 8, is characterized in that, described resistance R
1=R
2=1K ohm;
The scope that described microprocessor gathers voltage is 0-2.5V, and A/D conversion accuracy is minimum is 12, and sample rate is at least 1Kbit/s;
Described D/A conversion unit applies voltage to another utmost point of electric capacity, and voltage conversion range is 0-2.5V, and conversion accuracy is 10.
10. the detection method of a kind of Graphene capacitor biological sensor as claimed in claim 7, it is characterized in that, described capacitor electrode capacitance computation process: select time constant t=RC, it is 0.63 times that capacitance voltage is charged to D/A output voltage, performance graph constant computing time t according to measuring, obtains capacitor electrode capacitance according to formula C=t/R.
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| CN106383196A (en) * | 2016-10-31 | 2017-02-08 | 山东师范大学 | Biosensor of graphene/copper particle mixing structure and preparation method thereof |
| WO2018053932A1 (en) * | 2016-09-20 | 2018-03-29 | 德州学院 | Sensing device and method for detecting binding energy and binding kinetics among molecules |
| CN109468220A (en) * | 2018-11-15 | 2019-03-15 | 江南大学 | Graphene chip for microbial cell biological information real-time detection |
| RU2815247C1 (en) * | 2022-11-07 | 2024-03-12 | ООО "Нитридные кристаллы-Алюминий-Н" | Biosensor |
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