CN220525724U - Multifunctional electrochemical detection cell based on microelectrode array chip - Google Patents
Multifunctional electrochemical detection cell based on microelectrode array chip Download PDFInfo
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model discloses a multifunctional electrochemical detection cell based on a microelectrode array chip, which consists of a liquid storage tank and a chip; the liquid storage tank is positioned above the chip, and the lower end of the liquid storage tank is fixedly connected with one side surface of the chip; the chip comprises an electrode substrate, wherein the electrode substrate is square; two first working electrode groups and two second working electrode groups are arranged on the electrode substrate; a pair of electrodes are arranged between the first working electrode group and the second working electrode group, and the pair of electrodes are positioned on one side of the first working electrode group or the second working electrode group, and the length direction of the pair of electrodes is consistent with the length direction of the first working electrode group or the second working electrode group; a reference electrode is also provided between the counter electrode and the first working electrode set.
Description
Technical Field
The utility model relates to the technical field of electrochemical sensing detection, in particular to a multifunctional electrochemical detection cell based on a microelectrode array chip.
Background
Biomolecules have important regulatory roles in the vital activities of organisms, and are involved in many biochemical processes, including storing and transmitting genetic information, regulating biological/neural activities, small molecule/hormone transport, and catalyzing various biochemical reactions. The sensitive and real-time detection and analysis of biological small molecules has important significance in disease diagnosis and health management. In the past decades, electrochemical sensing technology has been developed as a powerful tool for detection and analysis of biomolecules due to the advantages of high response speed, high sensitivity, diversified electrode designs, integrability and the like.
Microelectrodes of dimensions on a microscale exhibit unique advantages compared to conventional electrodes, such as enhanced mass transport, faster transient response and lower detection destructive. A number of microelectrode arrangements combine to form a microelectrode array (MEA). The MEA can be constructed on a solid substrate through an electrochemical deposition or electroless plating process, and has the characteristics of small size, high integration level and the like. The size, morphology, structure and materials of the electrodes generally determine the electrochemical performance of the MEA. In addition, by modifying the electrode with specific recognition sites, functional molecules, or nanomaterials, the sensitivity and selectivity of the MEA can be improved. The MEA is used as an effective electrochemical sensing device and can promote the in-vivo and in-vitro biomolecule sensing research.
However, some MEAs currently being developed are often too compact in structural layout due to their small size, with the risk of interaction and cross-contamination between the electrodes. Furthermore, due to the small structural design, the MEA has substantially no integrated reference and counter electrode structures, which makes integration and portability a significant compromise. In addition, most MEAs have only a single working electrode layout, the detection targets are limited, and the functions are single. More importantly, most MEA configurations are based on precious metal materials, which are prone to biofouling, and the electrodes are not renewable, making their production cost high and practical.
Disclosure of Invention
Aiming at the defects existing in the prior art, the utility model aims to provide a multifunctional electrochemical detection cell based on a microelectrode array chip, so as to solve the problems of insufficient integration and portability, single function and non-reproducibility in the prior art.
The technical problems are solved, and the utility model adopts the following technical scheme:
the multifunctional electrochemical detection cell based on the microelectrode array chip comprises a liquid storage tank and a chip; the liquid storage tank is a liquid storage space surrounded by a baffle plate raised from the electrode arrangement surface of the chip; a plurality of working electrodes with different functions are arranged on the electrode arrangement surface of the chip; one end of the detection working electrode is positioned in the liquid storage tank, and the other end of the detection working electrode extends from the liquid storage tank to the edge of the chip and is connected with the contact through the electrode wire.
The liquid storage tank is arranged on the chip, and not only can be used for storing electrolyte and cleaning liquid, but also can be used for placing cells to be detected into the liquid storage tank for culturing. Moreover, the liquid storage tank is integrated with the chip, so that the whole detection tank is more compact in structure and convenient to carry and operate; on the other hand, the liquid storage tank on the chip stores the cleaning solvent, so that physical damage to the chip electrode caused by modes such as flowing flushing can be avoided, and the detection performance of the chip electrode is further influenced.
Preferably, the detection working electrode comprises two first working electrode groups and two second working electrode groups, and each of the first working electrode groups and the second working electrode groups comprises a plurality of working electrodes; the length direction of the first working electrode group is perpendicular to the length direction of the second working electrode group, but all working electrodes in the first working electrode group and the second working electrode group are not intersected. One end of the working electrode is positioned in the liquid storage tank, and the other end of the working electrode is connected with the contact through the electrode wire after extending out of the liquid storage tank. A counter electrode is arranged between the first working electrode group and the second working electrode group, and the counter electrode is positioned at one side of the first working electrode group or the second working electrode group, and the length direction of the counter electrode is consistent with the length direction of the first working electrode group or the second working electrode group; and a reference electrode is further arranged between the counter electrode and the first working electrode group, and an included angle between the length direction of the reference electrode and the length direction of the first working electrode group is an acute angle. Likewise, the counter electrode and the reference electrode, and the working electrode are not intersected. One end of the counter electrode and one end of the reference electrode are positioned in the liquid storage tank, and the other end of the counter electrode and the reference electrode extend out of the liquid storage tank and are connected with the contact through the electrode wire. The utility model further optimizes the arrangement of the electrodes on the chip, so that the chip structure layout is more compact, and the space is saved. Meanwhile, the counter electrode and the working electrode are arranged, so that a complete current measurement loop can be formed between the counter electrode and the working electrode, and a fixed reference potential can be provided and maintained for the chip by utilizing the reference electrode.
Preferably, the electrode base comprises a substrate, and a layer of gold nanoparticle/indium tin oxide (AnNPs/ITO) film is arranged on the substrate.
Preferably, in the first working electrode group and the second working electrode group, a plurality of working electrodes are disposed in parallel with each other.
Preferably, the working electrode comprises long electrodes and short electrodes, and the long electrodes and the short electrodes are alternately distributed, so that the chip structure layout is compact; the long and short electrodes can also be used as function distinguishing electrodes for different molecular detection, the long electrode can realize Uric Acid (UA) measurement through catalytic oxidation, and the short electrode can realize hydrogen peroxide (H) through catalytic reduction 2 O 2 ) And (5) measuring.
Preferably, the chip is square, and the side length of the chip is 3cm; the section of the liquid storage tank is circular, and the diameter of the liquid storage tank is 1.5cm; the axis of the liquid storage tank coincides with the central line of the chip.
Preferably, the microelectrodes are provided on the electrode substrate by laser etching.
The electrochemical detection cell based on the microelectrode array chip is prepared by the following method:
(1) Preparation of ITO film: cutting chemical glass (the size is 30mm multiplied by 30 mm), plating an ITO film on the surface of the chemical glass by a magnetron sputtering method, wherein the sputtering power is 10kW, the thickness is about 380nm, the resistance is 3-4 omega, and the light transmittance is more than or equal to 78%;
(2) Microelectrode array layout design: designing a microelectrode array layout by means of a computer-aided program, wherein the electrode line width is 0.2mm, the electrode line connecting point line width is 3mm, and 26 microelectrodes are included in total;
(3) And (3) laser etching: the electrode layout is led into a laser etching system for etching, and the laser etching parameters are as follows: laser wavelength 1064nm, laser working power 30% and etching rate 200mms -1 A plurality of ITO microelectrodes are formed on the electrode substrate.
(4) Preparation of Polydimethylsiloxane (PDMS) reservoirs: the mold was designed by means of a computer-aided program and the abrasive article was prepared by means of 3D printing. The silicone elastomer base was mixed with the curing agent in a ratio of 10:1 and degassed in a vacuum chamber. The polydimethylsiloxane PDMS mixture was then poured into a mold and cured in an oven at 70 ℃ for 1 hour;
(5) Assembling an electrochemical chip: and stripping the PDMS liquid storage tank, and combining the PDMS liquid storage tank with the ITO microelectrode array chip after high-power oxygen plasma treatment for 30 seconds to obtain the electrochemical detection tank. Wherein gold nanoparticles are modified on the microelectrodes by electrochemical deposition.
The electrochemical detection cell can be used for measuring biological small molecules, and concretely comprises the step of secreting H to cells 2 O 2 And oxidation of UA in urine.
After the electrochemical detection cell is used, the electrochemical detection cell can be cleaned by the following method: will H 2 O 2 Placing the chip in a liquid storage tank, and irradiating the chip for 2 hours under ultraviolet light to complete the regeneration of the electrochemical detection cell.
Compared with the prior art, the utility model has the following advantages:
1. according to the utility model, the liquid storage tank is arranged on the chip, and can be used for storing electrolyte and cleaning liquid, and cells to be detected can be placed in the liquid storage tank for culture; moreover, the liquid storage tank is integrated with the chip, so that the whole detection tank is more compact in structure and convenient to carry and operate; on the other hand, the liquid storage tank on the chip stores the cleaning solvent, so that physical damage to the chip electrode caused by modes such as flowing flushing can be avoided, and the detection performance of the chip electrode is further influenced.
2. The utility model integrates the working electrode, the counter electrode and the reference electrode miniature three-electrode system on the chip, and alternately distributes the working electrodes with different functions, thereby enhancing portability and functionality; the detection tank is integrated with the liquid storage tank, so that the structure of the whole detection tank is more compact; the nano structure of the microelectrode enables the chip to have a self-cleaning function, so that an extra cleaning procedure can be avoided, and the chip is simple, convenient and practical.
Drawings
FIG. 1 is a schematic diagram of a multifunctional electrochemical detection cell based on a microelectrode array chip according to the present utility model.
FIG. 2a is a schematic diagram of the preparation of a chip in the electrochemical detection cell; FIG. 2b is a schematic diagram of the structural layout thereof; FIG. 2c is a schematic illustration of the electrochemical detection cell; FIG. 2d is a physical diagram of the electrochemical detection cell.
FIG. 3a is an SEM image of the MEA and ESD element map in the electrochemical detection cell; figure 3b is an XRD pattern of MEA in the electrochemical detection cell.
FIG. 4a is a CV response curve of different MEAs in 5mM potassium ferricyanide solution; FIG. 4b is a CV response curve of MEA in the electrochemical detection cell scanned 5 consecutive turns in 5mM potassium ferricyanide solution; FIG. 4c is a schematic illustration of an MEA; FIG. 4d is the current response of 20 microelectrodes (1-20) in 5mM potassium ferricyanide solution.
FIG. 5a shows the chip pairs of different concentrations H in the electrochemical detection cell 2 O 2 Is a CV response curve of (2); FIG. 5b shows a chip in the electrochemical detection cell containing 2mM H 2 O 2 CV response curves at different scan rates in PBS, with the inset being the linear relationship between the reduction current and the scan rate; FIG. 5c is H 2 O 2 A detection mechanism of molecules at an electrochemical sensing interface; FIG. 5d shows the chip pairs of different concentrations H in the electrochemical detection cell 2 O 2 I-t response curve, inset: current response and H 2 O 2 Linear relationship of concentration (n=3); FIG. 5e shows a chip pair 500. Mu. M H in the electrochemical detection cell 2 O 2 And an i-t response curve for 5mM interferents; FIG. 4f shows a 500. Mu. M H electrode pair for a chip in a 6-group electrochemical test cell 2 O 2 I-t response curve of (c).
FIG. 6a shows cell attachment and H on chip in the electrochemical detection cell 2 O 2 A molecular monitoring schematic; FIG. 6b is a microscopic and fluorescent image of cells cultured on-chip for 12 hours in the electrochemical detection cell; FIG. 6c is an i-t response curve of the chip in the electrochemical detection cell under different stimulus conditions; FIG. 6d is a graph showing the current response of the chip in the electrochemical detection cell under different stimulus conditions.
FIG. 7a is a schematic diagram of detection of UA on a chip in the electrochemical detection cell; fig. 7b is a linear relationship between oxidation current and scan rate, inset: CV curves of chips in the electrochemical detection cell to 500 mu M UA under different scanning rates; fig. 7c is a graph showing the DPV response of the chip in the electrochemical detection cell to various concentrations UA, in inset: linear relation of current response to UA concentration (n=3); FIG. 7d is a graph showing the DPV response of a chip in the electrochemical detection cell to 500. Mu.M UA and 5mM interferents; FIG. 7e is a graph showing DPV response of 500. Mu.M UA for different microelectrode pairs; FIG. 7f is a graph showing the DPV response of the chip in the electrochemical test cell in two different urine samples, inset: the electrochemical detection cell and commercial detection kit measure UA concentration.
FIG. 8a is a schematic view of regeneration of an electrode after UV irradiation; FIG. 8b is a schematic diagram showing the mechanism of photocatalysis of a chip composite structure in the electrochemical detection cell; FIG. 8c is a graph showing CV response curves in potassium ferricyanide solution before passivation, after passivation and after UV irradiation of the chip for 2 hours in the electrochemical detection cell; fig. 8d is the corresponding peak current percentage (n=4).
In the figure: 1 to 20 are working electrodes; CE1 to CE4 are counter electrodes; RE 1-RE 2 are reference electrodes.
Detailed Description
The utility model will be further described with reference to the drawings and examples.
1. Multifunctional electrochemical detection cell based on microelectrode array chip
The electrochemical detection cell consists of a liquid storage tank and a chip, as shown in fig. 1. The liquid storage tank is a liquid storage space surrounded by a baffle plate raised from the electrode arrangement surface of the chip, so that the lower end of the liquid storage tank can be sealed by one side surface of the chip, and liquid can be poured into the liquid storage tank. The liquid storage tank is not only used for storing electrolyte liquid, but also can be used for placing cells to be detected into the liquid storage tank for culturing.
The electrochemical detection cell consists of a liquid storage tank and a chip; the liquid storage tank is a liquid storage space surrounded by a baffle plate raised from the electrode arrangement surface of the chip; a plurality of detection working electrodes are arranged on the electrode arrangement surface of the chip; one end of the detection working electrode is positioned in the liquid storage tank, and the other end of the detection working electrode extends from the liquid storage tank to the edge of the chip and is connected with the contact through the electrode wire. The liquid storage tank is arranged on the chip, and not only can be used for storing electrolyte and cleaning liquid, but also can be used for placing cells to be detected into the liquid storage tank for culturing. The liquid storage tank is integrated with the chip, so that the whole detection tank is more compact in structure and convenient to carry and operate; on the other hand, the liquid storage tank on the chip stores the cleaning solvent, so that the damage to the chip electrode caused by the modes of flowing flushing and the like is avoided, and the detection performance is further influenced.
In specific implementation, the detection working electrode includes two first working electrode groups and two second working electrode groups, where the first working electrode groups and the second working electrode groups each include a plurality of working electrodes, as shown in fig. 4c, where 1-5 and 11-15 are all the first working electrode groups, and 6-10 and 16-20 are all the second working electrode groups. The length direction of the first working electrode group is perpendicular to the length direction of the second working electrode group; one end of the working electrode is positioned in the liquid storage tank, and the other end of the working electrode is connected with the contact through the electrode wire after extending out of the liquid storage tank. The counter electrodes are arranged between the first working electrode group and the second working electrode group, as shown in FIG. 4c, CE 1-CE 4 are counter electrodes, which are positioned at one side of the first working electrode group or the second working electrode group and the length direction of which is equal to that of the first working electrode group or the second working electrode groupThe length direction of the groups is uniform. A reference electrode is further arranged between the counter electrode and the first working electrode group, as shown in fig. 4c, RE1 and RE2 are reference electrodes, and an included angle between the length direction of the reference electrode and the length direction of the first working electrode group is an acute angle; one end of the counter electrode and one end of the reference electrode are positioned in the liquid storage tank, and the other end of the counter electrode and the reference electrode extend out of the liquid storage tank and are connected with the contact through the electrode wire. Wherein the working electrode is AuNPs/ITO, the counter electrode is ITO, and the reference electrode is Ag/AgCl/ITO. The electrode base is a glass substrate, and a layer of ITO film is arranged on the glass substrate by a magnetron sputtering method. In the first working electrode group and the second working electrode group, a plurality of working electrodes are arranged in parallel to each other. The working electrode comprises long electrodes and short electrodes, and the long electrodes and the short electrodes are alternately distributed, so that the chip structure layout is compact, and the space is saved; the long electrode and the short electrode can also be used as function distinguishing electrodes for different molecular detection, the long electrode realizes UA measurement through catalytic oxidation, and the short electrode realizes H-passing through catalytic reduction 2 O 2 And (5) measuring. As shown in FIG. 4c, long electrodes 2, 4, 7, 9, 12, 14, 17 and 19 can be provided as working electrodes for UA measurement, and short electrodes 1, 3, 5, 6, 8, 10, 11, 13, 15, 16, 18 and 20 can be provided for H 2 O 2 And a working electrode for measurement. The chip is square, and the side length of the chip is 3cm. The square chip is convenient to cut and prepare, and is convenient to position during wiring and installation; the cross section of the liquid storage tank is circular, and the diameter of the liquid storage tank is 1.5cm. The round liquid storage tank has less material consumption and is convenient to clean, cleaning liquid can be poured into the round liquid storage tank, electrodes in the round liquid storage tank are soaked and cleaned, physical damage to the chip electrodes caused by flowing flushing and other modes is avoided, and the detection performance of the chip electrodes is further affected. The axis of the liquid storage tank coincides with the central line of the chip. The microelectrodes are arranged on the electrode substrate by laser etching.
2. Preparation method of electrochemical detection cell based on microelectrode array chip
In the utility model, the microelectrode is an ITO microelectrode, and the microelectrode forms an array on an electrode substrate, so that the chip is obtained. The chip is etched by laserThe preparation method has the characteristics of small size, easy operation, low cost and high yield. The specific preparation process is shown in figure 2a, firstly, a layer of ITO film is plated on the surface of the cleaned glass by using a magnetron sputtering method, the thickness is about 380nm, the resistance is 3-4Ω, and the light transmittance is more than or equal to 78%. Meanwhile, the layout of the microelectrode array is designed by means of a computer aided program, and the microelectrode array is led into a laser etching system for etching, so that a chip with a plurality of indium tin oxide microelectrode arrays on the surface is obtained. Wherein, the laser etching parameters are as follows: laser wavelength 1064nm, laser working power 30% and etching rate 200mms -1 。
The electrochemical detection cell comprises 1 chip and 1 liquid storage tank, the liquid storage tank is used for storing electrolyte liquid, the liquid storage tank is round in shape and has a diameter of 1.5cm (figure 2 b), and the height of the liquid storage tank is at least 1cm. The electrochemical reservoir is prepared by curing PDMS materials by molding. First, the reservoir mold is designed by computer-aided programming and prepared by means of 3D printing. The silicone elastomer base was mixed with the curing agent in a ratio of 10:1 and degassed in a vacuum chamber. The PDMS mixture was then poured into a mold and cured in an oven at 70 ℃ for 1 hour. The PDMS reservoirs were peeled off and ready to be bonded to the chip after 30 seconds of high power oxygen plasma treatment. Fig. 2c and 2d show a schematic and a physical diagram, respectively, of the electrochemical detection cell.
The chip was ultrasonically cleaned with acetone, ethanol, and a large amount of deionized water in sequence for 10 minutes, and blow-dried with nitrogen. 16 Working Electrodes (WE) and 2 Reference Electrodes (RE) were obtained using a droplet-on-thermosetting method, respectively, using an electrochemical deposition process. Wherein the working electrode is AuNPs/ITO, the counter electrode is ITO, and the reference electrode is Ag/AgCl/ITO. The reservoir is then assembled onto the chip to yield the electrochemical detection cell.
Characterization of the electrode morphology of the chip in the electrochemical detection cell, figure 3a shows the structural layout of the microelectrode array, after electrochemical deposition, a large number of nanoparticles are uniformly distributed on the electrode surface (figure 3 b). The Energy Dispersive Spectroscopy (EDS) element map clearly shows the presence of ln, O, au elements (fig. 3 c). In addition, the crystal structure of AuNPs/ITO is subjected to XRDAnd (5) analyzing. As shown In FIG. 3d, auNPs/ITO showed distinct and strong diffraction peaks at 38.8 °, 45.0 °, 65.1 °, 78.1 ° and 82.3 °, corresponding to In, respectively 2 O 3 The (211), (222), (400), (440) and (622) planes. In addition, diffraction peaks at 38.5 ° and 45.0 ° are due to the (111) and (200) crystal planes of AuNPs.
Characterization of the electrochemical performance of the chip in the electrochemical cell, fig. 4a shows that the prepared ITO MEA exhibits a pair of well-defined redox peaks in potassium ferricyanide solution, indicating successful construction of the three electrode system. The redox current is further enhanced after electrochemical deposition of the modified AuNPs. The AuNPs/ITO MEA had good redox peak overlap after 5 consecutive tests, with no obvious signs of decay (FIG. 4 b), indicating that the microelectrodes produced had excellent electrochemical stability. The microelectrode array is distributed as shown in FIG. 4c, with working electrodes numbered 1 through 20, of which microelectrodes numbered 2, 4, 7, 9, 12 and 14 are somewhat longer and the other somewhat shorter. The peak current values of oxidation of 20 working electrodes in potassium ferricyanide solution are shown in fig. 4d, and it can be seen that the Relative Standard Deviation (RSD) of the current values of both the long electrode and the short electrode is about 5%, and good reproducibility is exhibited. It can be seen that the prepared microelectrode array has satisfactory electrochemical testing performance.
3. Application of electrochemical detection cell
1. Secretion of H to cells 2 O 2 Reduction assay of (2)
FIG. 5a shows the electrochemical detection cell containing different concentrations of H 2 O 2 As can be seen, the CV response is H 2 O 2 The concentration increase is continuously enhanced, and the electrochemical detection cell is used for detecting H 2 O 2 Has good catalytic detection capability. FIG. 5b shows the electrochemical detection cell in the presence of 2mM H 2 O 2 CV curves of PBS at different scan rates, whose reduction current and scan rate are linear, indicate H 2 O 2 The catalytic reaction of molecules on the surface of a chip in the electrochemical detection cell is a surface adsorption control process. Here, H 2 O 2 The catalytic mechanism of the molecules on the chip surface is shown in FIG. 5c, involving H 2 O 2 Cracking, adsorbing and reducing. Wherein the OH intermediate adsorbed on the surface of the material plays an important role, and the adsorption of OH (OH (ads) ) And reduction is considered to be H 2 O 2 The rate limiting step of dissociation reduction, stronger OH adsorption can promote the cleavage of O-O bonds and subsequent reduction reactions. The electrochemical detection cell pair H was studied using the i-t test 2 O 2 Electrochemical detection properties of the molecules. Continuously injecting H with different concentrations into PBS solution at a working voltage of-0.6V 2 O 2 And a typical i-t response curve was recorded. As shown in fig. 5d, with H 2 O 2 The increase in concentration, the i-t response curve, gradually shifts toward negative current, indicating a gradual increase in reduction current. In the range of 10-500. Mu.M, the current response and H 2 O 2 There is a good linear relationship between concentrations (figure 5d inset). In addition, the selectivity of the electrochemical detection cell was investigated. As shown in FIG. 5e, 500 μ M H 2 O 2 While the addition of 5mM common interferents include Ascorbic Acid (AA), dopamine (DA), uric Acid (UA), glucose (Glu), sodium chloride (NaCl), urea (Ure), sodium nitrite (NaNO) 2 ) And ferric chloride (FeCl) 3 ) Without causing a significant current response. By recording different microelectrode pairs 500 mu M H 2 O 2 To study the reproducibility of the electrochemical detection cell. As shown in FIG. 5f, 6 different AuNPs/ITO microelectrode pairs 500 μ M H 2 O 2 The RSD of the current response of (a) was 6.65%, and the repeatability was good.
2. In situ monitoring of cell release H 2 O 2 Application study of (2)
Application of electrochemical detection cell to in situ real-time monitoring of cell secreted H 2 O 2 A molecule. The detection principle is shown in FIG. 6a, and PMA stimulates cells to produce H 2 O 2 Secreted H 2 O 2 Diffuse to the electrode surface, are catalytically reduced, generate electron transfer, and are converted into electrochemical signals. FIG. 6b shows cell microscopy and fluorescence development for 12 hours on an electrochemical detection cell chipThe color image clearly shows that a large number of cells are well attached and grown on the chip. The cells are directly attached and grown on the surface of the electrode, so that the loss of molecules from solution to the surface of the electrode can be well avoided, and timely capturing and detection are realized. FIG. 6c is a graph showing the i-t response of the chip in the electrochemical test cell under different stimulus conditions. As shown, in the cell-attached state, a pronounced current response (blue and pink lines) was recorded after PMA stimulation. In control experiments without cells, PMA stimulation did not elicit a significant current response (black and red). In addition, the current response obtained in the cell-attached state was significantly higher than that of the control experiment without cells (FIG. 6 d), indicating that the electrochemical detection cell successfully monitored the cell-secreted H 2 O 2 A molecule.
3. The electrochemical detection cell is used for the sensing detection of UA
The electrochemical detection cell is applied to the catalytic detection of UA molecules. H 2 O 2 The detection of the molecules is based on H 2 O 2 The catalytic reduction is carried out on the surface of the electrode, and the detection of the UA molecules is based on the catalytic oxidation of UA on the surface of the electrode, so that the versatility of the chip in the electrochemical detection cell can be demonstrated. Fig. 7a is a schematic diagram of detection of a UA molecule on a chip in the electrochemical detection cell, where the UA may bind to the electrode surface through a hydrogen atom of an amine group in an imidazole ring to induce n—h bond to become longer, and finally undergo cleavage rearrangement under the action of an applied voltage, losing protons and electrons, and undergoing an oxidation reaction. In order to investigate the electrochemical detection performance of the electrochemical detection cell on UA molecules, CV and DPV tests were performed. FIG. 7b is a graph showing the CV curve and the linear relationship between the oxidation current and the scan rate at different scan rates in PBS containing 500. Mu. MUA. The good linear relationship between the oxidation current and the scanning rate shows that the catalytic reaction of the UA molecules on the surface of the electrode is an adsorption control process. Fig. 7c is a graph of the DPV response of the electrochemical detection cell in PBS containing different concentrations of UA, and it can be seen that the DPV response increases significantly with increasing UA concentration, showing a good linear relationship in the range of 50-4000 μm. In addition, the choice of the electrochemical detection cell was further exploredSex and repeatability. As shown in fig. 7d, the electrochemical detection cell showed a significant DPV response to 500 μm UA, whereas no significant DPV response was observed for 5mM interferents including urea (Ure), glucose (Glu), sodium chloride (NaCl), creatinine (Cre), ascorbic Acid (AA) and guanine (Gua), with excellent selectivity. The RSD value of the DPV response of the 4 different microelectrode pairs to 500 μm UA was 3.24% (fig. 7 e), showing good reproducibility.
Finally, the chip in the electrochemical detection cell is applied to detection of UA molecules in actual urine. Here, two different urine samples were tested, as shown in fig. 7f, the UA signal in urine was clearly visible, and there was a difference in the measured UA concentration in the different urine samples, which may be due to individual differences. In addition, the detection result has good consistency with the detection result of a commercial UA detection kit (figure 7f inset), and the potential of the chip in the electrochemical detection cell in practical application is proved.
4. Renewable applications of the electrochemical detection cell
In the prior art, the non-reproducibility of the electrochemical sensor electrodes tends to increase the production cost. Therefore, the utility model introduces the photocatalysis technology to realize the renewable application of the electrode. The nano structure of the microelectrode enables the chip to have a self-cleaning function, so that an extra cleaning procedure can be avoided; meanwhile, the liquid storage tank on the chip can store the cleaning solvent, so that the damage to the chip electrode caused by the modes of flowing flushing and the like is avoided, and the detection performance is further affected. Specifically, the chip electrode of the electrochemical detection cell can realize self-cleaning of the electrode through ultraviolet light (UV) (fig. 8 a), and the light cleaning mechanism is shown in fig. 8b. FIG. 8c shows the CV curves of the test cells in 5mM potassium ferricyanide solution before passivation, after passivation and after UV irradiation. And the passivation rate and the regeneration rate of the electrode are obtained by taking the current response percentage as an ordinate and the passivation times as an abscissa. As shown in fig. 8d, the detection cell has a certain self-cleaning capability, and after multiple passivation and UV irradiation, the electrochemical activity can be restored to at least 83.95% of the initial value, which proves the renewable application performance of the detection cell.
Electrode structure the present utility model is not limited to the above embodiment, and it is within the scope of the present utility model to adopt the same or similar structure as the above embodiment of the present utility model.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present utility model and not for limiting the technical solution, and those skilled in the art should understand that modifications and equivalents may be made to the technical solution of the present utility model without departing from the spirit and scope of the present utility model, and all such modifications and equivalents are included in the scope of the claims.
Claims (7)
1. The multifunctional electrochemical detection cell based on the microelectrode array chip is characterized by comprising a liquid storage tank and a chip;
the liquid storage tank is a liquid storage space surrounded by a baffle plate raised from the electrode arrangement surface of the chip; a plurality of detection working electrodes are arranged on the electrode arrangement surface of the chip; one end of the detection working electrode is positioned in the liquid storage tank, and the other end of the detection working electrode extends from the liquid storage tank to the edge of the chip and is connected with the contact through the electrode wire.
2. The microelectrode array chip-based multifunctional electrochemical detection cell of claim 1, wherein the detection working electrode comprises two first working electrode sets and two second working electrode sets, each comprising a plurality of working electrodes; the length direction of the first working electrode group is perpendicular to the length direction of the second working electrode group;
a counter electrode is arranged between the first working electrode group and the second working electrode group, and the counter electrode is positioned at one side of the first working electrode group or the second working electrode group, and the length direction of the counter electrode is consistent with the length direction of the first working electrode group or the second working electrode group; and a reference electrode is further arranged between the counter electrode and the first working electrode group, and an included angle between the length direction of the reference electrode and the length direction of the first working electrode group is an acute angle.
3. The multifunctional electrochemical detection cell based on a microelectrode array chip according to claim 2, characterized in that the electrode base comprises a substrate on which a layer of gold nanoparticles/indium tin oxide film is provided.
4. The multi-functional electrochemical detection cell based on a microelectrode array chip according to claim 2, wherein the length directions of the plurality of working electrodes are arranged parallel to each other in the first working electrode group and the second working electrode group.
5. The multifunctional electrochemical detection cell based on the microelectrode array chip according to claim 2, wherein the working electrode comprises long electrodes and short electrodes, and the long electrodes and the short electrodes are alternately distributed.
6. The microelectrode array chip-based multifunctional electrochemical detection cell of claim 1, wherein the chip is square with a side length of 3cm; the section of the liquid storage tank is circular, and the diameter of the liquid storage tank is 1.5cm; the axis of the liquid storage tank coincides with the central line of the chip.
7. The multi-functional electrochemical detection cell based on a microelectrode array chip of claim 1, wherein the microelectrode is disposed on an electrode substrate by laser etching and electrochemical deposition.
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