CN112525969A - Preparation method of piezoelectric enhanced micro-fluidic photo-electrochemical sensor - Google Patents
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Abstract
The invention relates to a preparation method of a piezoelectric enhanced nano-array micro-fluidic control photo-electrochemical sensor for Procalcitonin (PCT) detection. The invention uses nano array ZnO/WO3The self-enhanced photocurrent assisted by the piezoelectric built-in electric field is obtained for the substrate material under the irradiation of pressure and visible light generated by fluid collision. The microfluidic biosensor consists of a microfluidic baseplate ITO conductive glass, a microfluidic chip and a screen printing microelectrode, wherein the microfluidic chip comprises an electrode groove for accommodating a counter electrode, a reference electrode, a working electrode, a sample inlet, a microchannel, a sample outlet and a microchannel; etching ITO conductive glass, and sequentially performing Ag/AgCI slurry screen printing and ZnO/WO3Modifying the nano material to obtain a base plate of a micro reference electrode and a micro working electrode; photoelectrochemistryThe three electrodes are integrated on the microfluidic biosensor, automatic detection can be realized by using the control of a pump, and an accurate detection result can be quickly obtained without artificial interference. The piezoelectric enhanced micro-fluidic photo-electrochemical sensor can realize rapid, efficient, sensitive and automatic detection of a septicemia marker procalcitonin.
Description
Technical Field
The invention relates to a piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor for septicemia marker detection.
Background
Septicemia refers to acute systemic infection caused by invasion of pathogenic bacteria or pathogenic bacteria into blood circulation, growth and reproduction in blood, and toxin production. Sepsis is classified into bacteremia without significant toxemia symptoms and sepsis with multiple abscesses, which if not rapidly controlled, can lead to tissue damage, organ dysfunction and even death. Therefore, early discovery and early treatment are of great significance for prevention and treatment of sepsis.
Procalcitonin is one of the best markers for diagnosing septicemia at present, and has important significance for early diagnosis of septicemia. At present, a plurality of methods for detecting septicemia markers are available, such as enzyme-linked immunoassay, electrochemiluminescence assay and the like, but the enzyme-linked immunoassay has methodological limiting factors such as low sensitivity, narrow linear range and the like; the electrochemical luminescence analysis method has wide detection linear range and simple operation, but is not easy to realize full automation. Therefore, the construction of a rapid, simple and sensitive detection method is of great significance.
The piezoelectric enhanced nano-array micro-fluidic photoelectrochemical sensor constructed by the invention is a detection technology for determining the concentration of an object to be detected based on a micro-fluidic sensing technology and photoelectric conversion, and has the advantages of small volume, less reagent consumption, automatic instrument, high sensitivity and the like. Therefore, a microfluidic-based photoelectrochemical sensor, i.e., a detection form in which separation of electron-hole pairs of a photoelectric material is excited by irradiation with a visible light LED, and the concentration of a substance to be detected is detected by conversion into an electric signal, has been attracting attention. The invention integrates the piezoelectric enhanced photoelectrochemical sensor technology on the microfluidic chip, and the piezoelectric enhanced photoelectrochemical sensor generates a certain built-in electric field by virtue of the piezoelectric property of the substrate material and the collision of microfluid on the material, has good separation effect on photo-generated electron hole pairs generated by the substrate material, and realizes the rapid, efficient and sensitive detection of the septicemia marker procalcitonin.
Disclosure of Invention
The invention aims to provide a simple, rapid, low-cost and high-sensitivity preparation method of a novel piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor, and the method is applied to detection of a septicemia marker procalcitonin.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a marker procalcitonin of septicemia is characterized in that the micro-fluidic biosensor consists of a micro-fluidic bottom plate ITO conductive glass, a micro-fluidic chip and a screen printing microelectrode, wherein the micro-fluidic chip comprises an electrode groove for arranging a counter electrode, a reference electrode, a working electrode, a sample inlet, a micro channel, a sample outlet and a micro channel;
the microfluidic baseplate is made of Indium Tin Oxide (ITO) conductive glass and used for being used as a screen printing microelectrode and being bonded with the baseplate of the microfluidic chip;
wherein, the screen printing microelectrode comprises a micro working electrode and a micro reference electrode;
wherein the sample inlet and the micro-channel comprise a septicemia marker antibody Ab1Sample inlet and microchannel, bovine serum albumin BSA sample inlet and microchannel, sepsis marker standard solution sample inlet and microchannel, sepsis marker antibody Ab2A marker injection port and a micro-channel, and a PBS solution injection port containing ascorbic acid and a micro-channel.
A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a septicemia marker comprises the following steps:
(1) designing and drawing a microfluidic channel graph by using computer design software AUTOCAD;
(2) drawing a mask by using the designed graph, and processing the microfluidic polydimethylsiloxane PDMS chip by using a standard soft lithography technology;
(3) sequentially ultrasonically cleaning 5 cm multiplied by 4 cm ITO conductive glass with acetone, ethanol and ultrapure water for 30 min, drying the ITO conductive glass by blowing with nitrogen, sequentially etching the cleaned ITO conductive glass, and screen-printing Ag/AgCI slurry to obtain bottom plates of a micro working electrode 1 and a micro reference electrode 2;
(4) dropping 20 mu L of 0.1-1.0 mol/L zinc nitrate and 0.1-1.0 mol/L hexamethylene tetramine mixed solution on a micro working electrode, airing at room temperature and annealing at 300 ℃ for 10 minutes, transferring the working electrode to 30 mL of aqueous solution containing 1.0 mol/L zinc nitrate and 1.0 mol/L hexamethylene tetramine, carrying out hydrothermal reaction at 95 ℃ for 9 hours, washing and drying, transferring the working electrode modified by the nano ZnO array to 30 mL of methanol solution containing 1.0-15.0 mg/mL tungsten hexachloride, reacting at 180 ℃ for 3 hours, washing and drying, annealing at 400 ℃ for 1 hour, continuously dripping 6-10 mu L and 0.5% chitosan solution, airing at room temperature, dripping 6-10 mu L and 0.5% glutaraldehyde solution, coupling, obtaining aldehyde ZnO/WO3A base plate of a modified micro-working electrode;
(5) the micro-fluidic chip prepared in the step (2) and the ZnO/WO prepared in the step (4)3The modified micro-working electrode bottom plate is processed by oxygen plasma together, and then the micro-fluidic chip is bonded with the bottom plate, thus completing the processPreparing a micro-fluidic chip;
(6) injecting 20 mug/mL of procalcitonin antibody Ab as septicemia marker at 20-40 muL/min through injection port 5 by using injection pump1To ZnO/WO in the working electrode tank of the micro-fluidic chip3Incubating in a refrigerator at 4 ℃ for 30-60 min, and injecting a buffer solution through a sample inlet 5 for washing to obtain ZnO/WO3/Ab1；
(7) Injecting bovine serum albumin BSA solution with the mass fraction of 0.1-1.0% into a working electrode tank by an injection pump through an injection port 6 at 20-40 muL/min3/Ab1To block unbound Ab on the electrode surface1Drying in a refrigerator at 4 ℃, and injecting a buffer solution from the sample inlet 6 for washing to obtain ZnO/WO3/Ab1/BSA；
(8) Injecting 10 pg/mL-100 ng/mL septicemia marker procalcitonin standard solution with different concentrations to the micro working electrode ZnO/WO at 20-40 muL/min through an injection port 7 by using an injection pump3/Ab1BSA, incubating in a refrigerator at 4 ℃ for 30-60 min, injecting a buffer solution into an injection port 7, and washing to obtain ZnO/WO3/Ab1/BSA/PCT；
(9) Injecting a 20 muL and 1.0-3.0 mg/mL secondary antibody marker solution (Ab) of silver nanoparticle-loaded silicon dioxide at 20-40 muL/min through a sample inlet 8 by using an injection pump2-Ag@SiO2) To the micro-working electrode ZnO/WO3/Ab1BSA/PCT, incubating in a refrigerator at 4 ℃ for 30-60 min, injecting a buffer solution into an injection port 8, and washing to obtain completely modified ZnO/WO3/Ab1/BSA/PCT/Ab2-Ag@SiO2A piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting septicemia markers.
Preferably, in the step (2), the micro-fluidic graph is used for drawing a mask, the diameter of a size electrode groove of the mask is 3000-4000 micrometers, the width of a connecting three-electrode micro-channel is 1000-2000 micrometers, the diameter of a sample inlet is 1000-1200 micrometers, the width of a sample inlet channel is 800-1000 micrometers, the diameter of a sample outlet is 1400-1600 micrometers, the width of a sample outlet channel is 1000-1200 micrometers, and the sample inlet and the sample outlet of the micro-fluidic channel are designed in a radian mode, so that smooth passing of liquid is guaranteed. The sample inlet and the sample outlet of the microfluidic channel are both designed in a radian manner, so that the liquid can smoothly pass through the microfluidic channel.
Preferably, in the step (3), the ITO conductive glass is a working electrode and is also a bottom plate of the microfluidic chip.
Preferably, in the step (5), the plasma treatment time is 40 s-60 s, and the finally bonded microfluidic chip is placed in an oven to be heated for 10 minutes at 80 ℃, so that the bonding between the chips is firmer.
Advantageous results of the invention
(1) The piezoelectric enhanced nano-array micro-fluidic control photoelectrochemical sensor prepared by the invention can overcome the defects of large electrolyte demand, poor repeatability, short service life and the like of the traditional photoelectrochemical sensor, can quantitatively detect small molecules and proteins by photoelectrochemistry, and has wide application prospect.
(2) Nano array ZnO/WO synthesized by the invention3The electrode combines the piezoelectric property and the photoelectric property of the material, effectively accelerates the electron transfer rate of an electrode interface, and further improves the signal stability and the reproducibility of the sensor.
(3) The micro-fluidic biosensor prepared by the invention integrates and reduces the micro-working electrode, the micro-reference electrode and the counter electrode on the micro-fluidic sensor, utilizes the control of the pump to realize the automatic detection of the sensor, and can quickly obtain an accurate detection result without artificial interference.
(4) The invention adopts the self-assembled LED as an excitation light source to realize the sensitive detection of the septicemia marker procalcitonin PCT, and provides important basis and technical breakthrough for realizing the piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor on the microfluidic chip.
Drawings
FIG. 1 is a schematic diagram of a microfluidic sensor provided by the present invention;
description of the reference numerals
1 a micro-working electrode; 2 micro reference electrode groove; 3 pairs of electrode grooves; 4, a sample outlet and a micro-channel; 5 septicemia marker antibody Ab1A sample inlet and a microchannel; 6 bovine serum albumin BSA sample inlet and micro-channel; 7, a procalcitonin standard solution sample inlet and a micro-channel; a sample inlet and a micro-channel of a secondary antibody marker solution of 8 silver nano-particle loaded silicon dioxide; 9 PBS solution injection port containing ascorbic acid and microchannel.
Detailed Description
Example 1
A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a septicemia marker comprises the following preparation steps:
(1) designing and drawing a microfluidic channel graph by using computer design software AUTOCAD;
(2) drawing a mask by using the designed graph, and processing the microfluidic polydimethylsiloxane PDMS chip by using a standard soft lithography technology;
(3) sequentially ultrasonically cleaning 5 cm multiplied by 4 cm ITO conductive glass with acetone, ethanol and ultrapure water for 30 min, drying the ITO conductive glass by blowing with nitrogen, sequentially etching the cleaned ITO conductive glass, and screen-printing Ag/AgCI slurry to obtain bottom plates of a micro working electrode 1 and a micro reference electrode 2;
(4) dripping 0.1 mol/L zinc nitrate and 0.1 mol/L hexamethylene tetramine mixed solution of 20 mu L on a micro working electrode, airing at room temperature and annealing at 300 ℃ for 10 minutes, transferring the working electrode to 30 mL aqueous solution containing 1.0 mol/L zinc nitrate and 1.0 mol/L hexamethylene tetramine, carrying out hydrothermal reaction at 95 ℃ for 9 hours, transferring the working electrode modified by the nano ZnO array to 30 mL methanol solution containing 1.0 mg/mL tungsten hexachloride after washing and drying, reacting at 180 ℃ for 3 hours, washing and drying, annealing at 400 ℃ for 1 hour, continuously dripping 6 mu L and 0.5% chitosan solution, airing at room temperature, dripping 6 mu L and 0.5% glutaraldehyde solution for coupling, and obtaining the ZnO/WO3A base plate of a modified micro-working electrode;
(5) the micro-fluidic chip prepared in the step (2) and the ZnO/WO prepared in the step (4)3Modified micro-working electrode backplaneCarrying out oxygen plasma treatment, and then bonding the microfluidic chip with the bottom plate to complete the preparation of the microfluidic chip;
(6) injecting 20 mug/mL of procalcitonin antibody Ab as septicemia marker at 20 muL/min through injection port 5 by using injection pump1To ZnO/WO in the working electrode tank of the micro-fluidic chip3Incubating in a refrigerator at 4 ℃ for 30 min, and injecting a buffer solution through a sample inlet 5 for washing to obtain ZnO/WO3/Ab1；
(7) Injecting bovine serum albumin BSA solution with the mass fraction of 0.1-1.0% into a working electrode tank at 20 muL/min by using an injection pump through an injection port 63/Ab1To block unbound Ab on the electrode surface1Drying in a refrigerator at 4 ℃, and injecting a buffer solution from the sample inlet 6 for washing to obtain ZnO/WO3/Ab1/BSA；
(8) Injecting 10 pg/mL-100 ng/mL septicemia marker procalcitonin standard solution with different concentrations to a micro-working electrode ZnO/WO at 20 muL/min through an injection port 7 by using an injection pump3/Ab1BSA, incubating in a refrigerator at 4 ℃ for 30 min, injecting a buffer solution into an injection port 7, and washing to obtain ZnO/WO3/Ab1/BSA/PCT；
(9) Injecting a 20 muL and 1.0-3.0 mg/mL secondary antibody marker solution (Ab) of silver nanoparticle-loaded silicon dioxide at 20-40 muL/min through a sample inlet 8 by using an injection pump2-Ag@SiO2) To the micro-working electrode ZnO/WO3/Ab1BSA/PCT, incubation in refrigerator at 4 deg.C for 30 min, injecting buffer solution into injection port 8, and washing to obtain completely modified ZnO/WO3/Ab1/BSA/PCT/Ab2-Ag@SiO2A piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting septicemia markers.
Example 2
A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a septicemia marker comprises the following preparation steps:
(1) designing and drawing a microfluidic channel graph by using computer design software AUTOCAD;
(2) drawing a mask by using the designed graph, and processing the microfluidic polydimethylsiloxane PDMS chip by using a standard soft lithography technology;
(3) sequentially ultrasonically cleaning 5 cm multiplied by 4 cm ITO conductive glass with acetone, ethanol and ultrapure water for 30 min, drying the ITO conductive glass by blowing with nitrogen, sequentially etching the cleaned ITO conductive glass, and screen-printing Ag/AgCI slurry to obtain bottom plates of a micro working electrode 1 and a micro reference electrode 2;
(4) dripping 0.5 mol/L zinc nitrate and 0.5 mol/L hexamethylene tetramine mixed solution of 20 mu L on a micro working electrode, airing at room temperature and annealing at 300 ℃ for 10 minutes, transferring the working electrode to 30 mL aqueous solution containing 1.0 mol/L zinc nitrate and 1.0 mol/L hexamethylene tetramine, carrying out hydrothermal reaction at 95 ℃ for 9 hours, transferring the working electrode modified by the nano ZnO array to 30 mL methanol solution containing 8.0 mg/mL tungsten hexachloride after washing and drying, reacting at 180 ℃ for 3 hours, washing and drying, annealing at 400 ℃ for 1 hour, continuously dripping 8 mu L and 0.5% chitosan solution, airing at room temperature, dripping 8 mu L and 0.5% glutaraldehyde solution for coupling, and obtaining the ZnO/WO3A base plate of a modified micro-working electrode;
(5) the micro-fluidic chip prepared in the step (2) and the ZnO/WO prepared in the step (4)3Carrying out oxygen plasma treatment on the modified micro-working electrode base plate, and then bonding the micro-fluidic chip with the base plate to finish the preparation of the micro-fluidic chip;
(6) injecting 20 mug/mL of procalcitonin antibody Ab as septicemia marker at 30 muL/min through injection port 5 by using injection pump1To ZnO/WO in the working electrode tank of the micro-fluidic chip3Incubating in a refrigerator at 4 ℃ for 45 min, and injecting a buffer solution through a sample inlet 5 for washing to obtain ZnO/WO3/Ab1；
(7) Injecting bovine serum albumin BSA solution with the mass fraction of 0.1-1.0% into a working electrode tank by an injection pump through an injection port 6 at 20-40 muL/min3/Ab1To seal the unjunction on the surface of the electrodeSynab1Drying in a refrigerator at 4 ℃, and injecting a buffer solution from the sample inlet 6 for washing to obtain ZnO/WO3/Ab1/BSA；
(8) Injecting 10 pg/mL-100 ng/mL septicemia marker procalcitonin standard solution with different concentrations to the micro working electrode ZnO/WO at 20-40 muL/min through an injection port 7 by using an injection pump3/Ab1BSA, incubating in a refrigerator at 4 ℃ for 45 min, injecting a buffer solution into an injection port 7, and washing to obtain ZnO/WO3/Ab1/BSA/PCT；
(9) Injecting a 20 muL and 2.0 mg/mL secondary antibody marker solution (Ab) of silver nanoparticle-loaded silicon dioxide at 20-40 muL/min through a sample inlet 8 by using an injection pump2-Ag@SiO2) To the micro-working electrode ZnO/WO3/Ab1BSA/PCT, incubation in refrigerator at 4 deg.C for 45 min, injecting buffer solution into injection port 8, and washing to obtain completely modified ZnO/WO3/Ab1/BSA/PCT/Ab2-Ag@SiO2A piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting septicemia markers.
Example 3
A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a septicemia marker comprises the following preparation steps:
(1) designing and drawing a microfluidic channel graph by using computer design software AUTOCAD;
(2) drawing a mask by using the designed graph, and processing the microfluidic polydimethylsiloxane PDMS chip by using a standard soft lithography technology;
(3) sequentially ultrasonically cleaning 5 cm multiplied by 4 cm ITO conductive glass with acetone, ethanol and ultrapure water for 30 min, drying the ITO conductive glass by blowing with nitrogen, sequentially etching the cleaned ITO conductive glass, and screen-printing Ag/AgCI slurry to obtain bottom plates of a micro working electrode 1 and a micro reference electrode 2;
(4) dripping 0.5 mol/L zinc nitrate and 0.5 mol/L hexamethylene tetramine mixed solution of 20 mu L on a micro-working electrode, and airing at room temperatureAnnealing at 300 ℃ for 10 minutes, transferring the working electrode to 30 mL of aqueous solution containing 1.0 mol/L of zinc nitrate and 1.0 mol/L of hexamethylenetetramine, carrying out hydrothermal reaction at 95 ℃ for 9 hours, washing and drying, transferring the nano ZnO array modified working electrode to 30 mL of methanol solution containing 15.0 mg/mL of tungsten hexachloride, reacting at 180 ℃ for 3 hours, washing and drying, annealing at 400 ℃ for 1 hour, continuously dripping 10 muL and 0.5% of chitosan solution, airing at room temperature, dripping 10 muL and 0.5% of glutaraldehyde solution for coupling, and obtaining aldehyde-based ZnO/WO3A base plate of a modified micro-working electrode;
(5) the micro-fluidic chip prepared in the step (2) and the ZnO/WO prepared in the step (4)3Carrying out oxygen plasma treatment on the modified micro-working electrode base plate, and then bonding the micro-fluidic chip with the base plate to finish the preparation of the micro-fluidic chip;
(6) injecting 20 mug/mL of procalcitonin antibody Ab as septicemia marker at 40 muL/min through injection port 5 by using injection pump1To ZnO/WO in the working electrode tank of the micro-fluidic chip3Incubating in a refrigerator at 4 ℃ for 60 min, and injecting a buffer solution through a sample inlet 5 for washing to obtain ZnO/WO3/Ab1；
(7) Injecting bovine serum albumin BSA solution with the mass fraction of 1.0% into a working electrode tank through an injection port 6 by using an injection pump at 20-40 mu L/min3/Ab1To block unbound Ab on the electrode surface1Drying in a refrigerator at 4 ℃, and injecting a buffer solution from the sample inlet 6 for washing to obtain ZnO/WO3/Ab1/BSA；
(8) Injecting 10 pg/mL-100 ng/mL septicemia marker procalcitonin standard solution with different concentrations to a micro-working electrode ZnO/WO at 40 muL/min through an injection port 7 by using an injection pump3/Ab1BSA, incubating in a refrigerator at 4 ℃ for 60 min, injecting a buffer solution into an injection port 7, and washing to obtain ZnO/WO3/Ab1/BSA/PCT；
(9) Injecting a 20 muL and 3.0 mg/mL secondary antibody marker solution of silver nanoparticle-loaded silicon dioxide at 20-40 muL/min by using an injection pump through a sample inlet 8 (Ab2-Ag@SiO2) To the micro-working electrode ZnO/WO3/Ab1BSA/PCT, incubation in refrigerator at 4 deg.C for 60 min, injecting buffer solution into injection port 8, and washing to obtain completely modified ZnO/WO3/Ab1/BSA/PCT/Ab2-Ag@SiO2A piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting septicemia markers.
Example 4
The diameter of an electrode groove of the micro-fluidic micro-channel with the size of the size is 3000 micrometers, the width of a connecting three-electrode micro-channel is 1000 micrometers, the diameter of a sample inlet is 1000 micrometers, the width of a sample inlet channel is 800 micrometers, the diameter of a sample outlet is 1400 micrometers, the width of the sample outlet channel is 1000 micrometers, and the sample inlet and the sample outlet of the micro-fluidic channel are both designed with radians, so that the liquid can smoothly pass through the micro-fluidic channel.
Example 5
The diameter of the electrode groove of the micro-fluidic micro-channel with the size of the size is 3500 mu m, the width of the connected three-electrode micro-channel is 1500 mu m, the diameter of the sample inlet is 1100 mu m, the width of the sample inlet channel is 900 mu m, the diameter of the sample outlet is 1500 mu m, the width of the sample outlet channel is 1100 mu m, and the sample inlet and the sample outlet of the micro-fluidic channel are both designed with radians to ensure that liquid smoothly passes through.
Example 6
The diameter of the micro-fluidic micro-channel size electrode groove is 4000 micrometers, the width of the connected three-electrode micro-channel is 2000 micrometers, the diameter of a sample inlet is 1200 micrometers, the width of a sample channel is 1000 micrometers, the diameter of a sample outlet is 1600 micrometers, the width of the sample outlet channel is 1200 micrometers, and the sample inlet and the sample outlet of the micro-fluidic channel are both designed with radians, so that the smooth passing of liquid is ensured.
Example 7
The steps of the piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor for detecting procalcitonin are as follows:
(1) the test is carried out in a three-electrode system by using an electrochemical workstation, a counter electrode is inserted into an electrode groove 3, a PBS solution with pH =7.4 and 0.1 mol/L ascorbic acid is injected through an injection port 9 and fills the electrode groove, and the test is carried out under the irradiation of an LED lamp;
(2) detecting procalcitonin by a time-current method, setting the voltage to be 0V, and running for 200 s;
(3) when the background current tends to be stable, turning on the lamp every 15 s for continuously irradiating for 15 s, then recording the change of photocurrent, and drawing a working curve;
(4) and replacing the procalcitonin standard solution with the serum sample solution, and checking the detection result through a working curve.
Example 8
The steps of the piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor for detecting procalcitonin are as follows:
(1) the test is carried out in a three-electrode system by using an electrochemical workstation, a counter electrode is inserted into an electrode groove 3, a PBS solution with pH =7.4 and 0.1 mol/L ascorbic acid is injected through an injection port 9 and fills the electrode groove, and the test is carried out under the irradiation of an LED lamp;
(2) detecting procalcitonin by a time-current method, setting the voltage to be 0V, and running for 200 s;
(3) when the background current tends to be stable, turning on the lamp for 20 s every 20 s, then recording the change of photocurrent, and drawing a working curve;
(4) and replacing the procalcitonin standard solution with the serum sample solution, and checking the detection result through a working curve.
Example 9
The steps of the piezoelectric enhanced nano-array microfluidic photoelectrochemical sensor for detecting procalcitonin are as follows:
(1) the test is carried out in a three-electrode system by using an electrochemical workstation, a counter electrode is inserted into an electrode groove 3, a PBS solution with pH =7.4 and 0.1 mol/L ascorbic acid is injected through an injection port 9 and fills the electrode groove, and the test is carried out under the irradiation of an LED lamp;
(2) detecting procalcitonin by a time-current method, setting the voltage to be 0V, and running for 200 s;
(3) when the background current tends to be stable, turning on the lamp every 25 s for continuously irradiating for 25 s, then recording the change of photocurrent, and drawing a working curve;
(4) and replacing the procalcitonin standard solution with the serum sample solution, and checking the detection result through a working curve.
Embodiment 10 the detection range of the piezoelectric enhanced nano-array microfluidic photoelectric chemical sensor for procalcitonin as a sepsis marker is 10 pg/mL to 100 ng/mL, and the detection limit is 2.6 pg/mL; simple, rapid, highly sensitive and specific detection can be realized.
Claims (7)
1. A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a marker Procalcitonin (PCT) of septicemia is characterized in that the micro-fluidic biosensor consists of a micro-fluidic bottom plate ITO conductive glass, a micro-fluidic chip and a screen printing microelectrode, wherein the micro-fluidic chip comprises an electrode groove for arranging a counter electrode, a reference electrode, a working electrode, a sample inlet, a micro-channel, a sample outlet and a micro-channel;
the microfluidic baseplate is made of Indium Tin Oxide (ITO) conductive glass and used for being used as a screen printing microelectrode and being bonded with the baseplate of the microfluidic chip;
wherein, the screen printing microelectrode comprises a micro working electrode and a micro reference electrode;
wherein the sample inlet and the micro-channel comprise procalcitonin antibody Ab1Sample inlet and microchannel, bovine serum albumin BSA sample inlet and microchannel, procalcitonin standard solution sample inlet and microchannel, procalcitonin antibody Ab2A marker injection port and a micro-channel, and a PBS solution injection port containing ascorbic acid and a micro-channel.
2. A preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting a septicemia marker is characterized in that the preparation steps of the piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor are as follows:
(1) designing and drawing a microfluidic channel graph by using computer design software AUTOCAD;
(2) drawing a mask by using the designed graph, and processing the microfluidic polydimethylsiloxane PDMS chip by using a standard soft lithography technology;
(3) sequentially ultrasonically cleaning 5 cm multiplied by 4 cm ITO conductive glass with acetone, ethanol and ultrapure water for 30 min, drying the ITO conductive glass by blowing with nitrogen, sequentially etching the cleaned ITO conductive glass, and screen-printing Ag/AgCI slurry to obtain bottom plates of a micro working electrode 1 and a micro reference electrode 2;
(4) dropping 20 mu L of 0.1-1.0 mol/L zinc nitrate and 0.1-1.0 mol/L hexamethylene tetramine mixed solution on a micro working electrode, airing at room temperature and annealing at 300 ℃ for 10 minutes, transferring the working electrode to 30 mL of aqueous solution containing 1.0 mol/L zinc nitrate and 1.0 mol/L hexamethylene tetramine, carrying out hydrothermal reaction at 95 ℃ for 9 hours, washing and drying, transferring the working electrode modified by the nano ZnO array to 30 mL of methanol solution containing 1.0-15.0 mg/mL tungsten hexachloride, reacting at 180 ℃ for 3 hours, washing and drying, annealing at 400 ℃ for 1 hour, continuously dripping 6-10 mu L and 0.5% chitosan solution, airing at room temperature, dripping 6-10 mu L and 0.5% glutaraldehyde solution, coupling, obtaining aldehyde ZnO/WO3A base plate of a modified micro-working electrode;
(5) the micro-fluidic chip prepared in the step (2) and the ZnO/WO prepared in the step (4)3Carrying out oxygen plasma treatment on the modified micro-working electrode base plate, and then bonding the micro-fluidic chip with the base plate to finish the preparation of the micro-fluidic chip;
(6) injecting 20 mug/mL of procalcitonin antibody Ab as septicemia marker at 20-40 muL/min through injection port 5 by using injection pump1To ZnO/WO in the working electrode tank of the micro-fluidic chip3Incubating in a refrigerator at 4 ℃ for 30-60 min, and injecting a buffer solution through a sample inlet 5 for washing to obtain ZnO/WO3/Ab1；
(7) Injecting bovine serum albumin BSA solution with the mass fraction of 0.1-1.0% into a working electrode tank by an injection pump through an injection port 6 at 20-40 muL/min3/Ab1To block unbound Ab on the electrode surface1Drying in a refrigerator at 4 ℃, and injecting a buffer solution from the sample inlet 6 for washing to obtain ZnO/WO3/Ab1/BSA；
(8) Injecting 10 pg/mL-100 ng/mL septicemia marker procalcitonin standard solution with different concentrations to the micro working electrode ZnO/WO at 20-40 muL/min through an injection port 7 by using an injection pump3/Ab1BSA, incubating in a refrigerator at 4 ℃ for 30-60 min, injecting a buffer solution into an injection port 7, and washing to obtain ZnO/WO3/Ab1/BSA/PCT；
(9) Injecting a 20 muL and 1.0-3.0 mg/mL secondary antibody marker solution (Ab) of silver nanoparticle-loaded silicon dioxide at 20-40 muL/min through a sample inlet 8 by using an injection pump2-Ag@SiO2) To the micro-working electrode ZnO/WO3/Ab1BSA/PCT, incubating in a refrigerator at 4 ℃ for 30-60 min, injecting a buffer solution into an injection port 8, and washing to obtain completely modified ZnO/WO3/Ab1/BSA/PCT/Ab2-Ag@SiO2A piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor, in particular to a preparation method of a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for detecting septicemia markers.
3. The preparation method of the piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for septicemia marker detection according to claim 2, wherein the diameter of the micro-fluidic micro-channel size electrode groove is 3000-4000 micrometers, the width of the three-electrode micro-channel is 1000-2000 micrometers, the diameter of the sample inlet is 1000-1200 micrometers, the width of the sample inlet channel is 800-1000 micrometers, the diameter of the sample outlet is 1400-1600 micrometers, the width of the sample outlet channel is 1000-1200 micrometers, and the sample inlet and the sample outlet of the micro-fluidic channel are both designed with radian to ensure smooth liquid passage.
4. The method for preparing a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for sepsis marker detection according to claim 2, wherein the photoelectrochemical three electrodes are integrated on the miniature micro-fluidic sensor.
5. The method according to claim 2, wherein the photoelectrochemical sensor is signal-enhanced by applying pressure to the nanoarray through microfluidic flow.
6. The method for preparing a piezoelectric enhanced nano-array micro-fluidic photo-electrochemical sensor for sepsis marker detection according to claim 2, wherein the sepsis marker is procalcitonin.
7. The photoelectrochemical microfluidic biosensor of claim 2, which detects procalcitonin, comprising the steps of:
(1) the test is carried out by using an electrochemical workstation in a three-electrode system, a counter electrode is inserted into an electrode groove 3, a Tris-HCI solution with the pH =7.4 and 0.1 mol/L is injected through an injection port 9, the electrode groove is filled, and the test is carried out under the irradiation of an LED lamp;
(2) detecting procalcitonin by a time-current method, setting the voltage to be 0V, and running for 200 s;
(3) when the background current tends to be stable, turning on the lamp every 15-25 s for continuously irradiating for 15-25 s, then recording the change of photocurrent, and drawing a working curve;
(4) and replacing the procalcitonin standard solution with the serum sample solution, and checking the detection result through a working curve.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110297023A (en) * | 2019-07-09 | 2019-10-01 | 济南大学 | A kind of preparation method and application for the self-reinforcing optical electro-chemistry immunosensor that detection Procalcitonin electrochemical catalysis is assisted |
| CN110487873A (en) * | 2019-09-19 | 2019-11-22 | 济南大学 | A kind of preparation method of the micro-fluidic optical electro-chemistry sensor for the detection of heart failure marker B-typeNatriuretic Peptide |
| CN110501393A (en) * | 2019-09-10 | 2019-11-26 | 济南大学 | A preparation method of photoelectrochemical immunosensor for detecting procalcitonin |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110297023A (en) * | 2019-07-09 | 2019-10-01 | 济南大学 | A kind of preparation method and application for the self-reinforcing optical electro-chemistry immunosensor that detection Procalcitonin electrochemical catalysis is assisted |
| CN110501393A (en) * | 2019-09-10 | 2019-11-26 | 济南大学 | A preparation method of photoelectrochemical immunosensor for detecting procalcitonin |
| CN110487873A (en) * | 2019-09-19 | 2019-11-22 | 济南大学 | A kind of preparation method of the micro-fluidic optical electro-chemistry sensor for the detection of heart failure marker B-typeNatriuretic Peptide |
Non-Patent Citations (2)
| Title |
|---|
| GUANG-ZHU CHEN ET AL.: "Electrochemical Immunoassay of Escherichia coli O157:H7 Using Ag@SiO2 Nanoparticles as Labels", 《JOURNAL OF ANALYTICAL METHODS IN CHEMISTRY》 * |
| YING CHEN ET AL.: "Polarization-Enhanced direct Z-scheme ZnO-WO3-x nanorod arrays for efficient piezoelectric-photoelectrochemical Water splitting", 《APPLIED CATALYSIS B: ENVIRONMENTAL》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113777143A (en) * | 2021-09-16 | 2021-12-10 | 哈尔滨工业大学（深圳） | A multi-channel integrated array photoelectrochemical sensing electrode and its preparation method and application |
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