CN114487055B - Multipath closed bipolar electrochemical luminescence chip and application thereof in detection sensing - Google Patents
Multipath closed bipolar electrochemical luminescence chip and application thereof in detection sensing Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- G—PHYSICS
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Abstract
The invention discloses a multipath closed bipolar electrochemical luminescence chip and application thereof in detection sensing, wherein the chip comprises an integrated bipolar electrode, a driving electrode anode, a driving electrode cathode, a supporting channel and a reporting channel; one end of the integrated bipolar electrode is a bipolar electrode cathode; the other end of the integrated bipolar electrode is divided into a plurality of parallel branches; the parallel branches cross more than one reporting channel; the part of the parallel branch which is arranged in the report channel is the bipolar electrode anode. In the chip, the serial bipolar electrode anode is connected with the same bipolar electrode cathode, so that the bipolar electrode cathode modification process is reduced, and multiple targets from different sample sources can be detected. In terms of the current flowing through the closed bipolar electrode, the magnitude of the current passing through each bipolar electrode anode is essentially the same, which is more advantageous for examining the lighting conditions of different areas to evaluate the accuracy of the detection.
Description
Technical Field
The invention belongs to the field of microfluidic analysis, and particularly relates to a multi-channel closed bipolar electrochemical luminescence chip and application thereof in detection and sensing.
Background
The microfluidic technology aims at integrating the behaviors of sample introduction, sample treatment, detection and the like on a microchip. Compared with the traditional detection method and detection instrument, the microfluidic chip has the characteristics of fluid controllability, low consumption of samples and reagents, high analysis speed and the like. In recent years, the development of closed bipolar electrodes combined with electrochemiluminescence has been diversified, and from single detection to high-flux detection, multiplex detection and the like, closed bipolar electrodes are often designed into different shapes and sizes on a microfluidic chip to realize different detection requirements. The closed bipolar electrode electrochemiluminescence microfluidic chip has potential to provide a rapid analysis tool for fields such as in-vitro diagnosis, environmental monitoring, food inspection and the like.
Most chips for multiplex or high throughput detection employ an array microfluidic chip comprising a plurality of bipolar electrodes and reaction cells connected in parallel.
The closed bipolar electrode anodes correspond to different bipolar electrode cathodes, and the difference of electrode manufacturing and modification processes can adversely affect signals of the bipolar electrode anodes, so that detection errors are increased. If several bipolar electrode anodes are located in the same reporting channel, the liquid consumption is large during detection, and it is difficult to perform multi-target detection of different sample sources. If a plurality of bipolar electrode anodes are positioned in different report tanks, the accurate measurement needs to be added with liquid for many times, and the operation is complex. In addition, the number and the size of the bipolar electrodes arranged in a radial manner are greatly influenced by the circumferential angle and the radius of the circle.
Disclosure of Invention
The invention aims to provide a multi-path closed bipolar electrochemical luminescence chip and application thereof in detection sensing, and the chip aims to solve the following defects in the prior art:
1. in the existing array type micro-fluidic chip, a plurality of closed bipolar electrodes are independently and parallelly arranged in a reaction channel, pretreatment is needed on each cathode or anode of the bipolar electrodes, the process is tedious and time-consuming, and the detection is adversely affected to a certain extent.
The serially connected closed bipolar electrode anodes are positioned in different reporting channels and connected with the same bipolar electrode cathode, so that the bipolar electrode cathode modification process is reduced, and multiple targets with different sample sources can be detected. In terms of the current flowing through the closed bipolar electrode, the magnitude of the current passing through each bipolar electrode anode is essentially the same, which is more advantageous for examining the lighting conditions of different areas to evaluate the accuracy of the detection. In addition, when the nanomaterial is modified at the bipolar electrode anode to promote electron transfer and amplify an electrochemical luminescence signal, modification of the tandem structure is simpler (only the anode closest to the bipolar electrode cathode needs to be modified), while the parallel structure needs to modify all anodes.
2. In the traditional unit detection, only single data is generated in single detection, and the detection efficiency and the accuracy are low. In the invention, a plurality of serially connected bipolar electrode anodes are distributed in parallel and symmetrically along the electrode part between the bipolar electrode cathode and the anode, the closed bipolar electrode comprises a cathode and a plurality of serial and parallel anodes, and a plurality of results can be simultaneously generated in one-time detection no matter the cathode sensing or the anode sensing, so that the detection efficiency and the accuracy are greatly improved.
3. The current micro-fluidic chip closed bipolar electrode technology has a single power supply mode, and cannot well meet the requirements of multi-element detection and accurate detection. In the invention, different driving electrodes are simultaneously or individually powered to perform electrochemical luminescence detection of a plurality of series-parallel closed bipolar electrodes, thereby meeting the requirements of simultaneous or non-simultaneous multi-element and accurate detection.
4. Generally, when a plurality of parallel closed bipolar electrodes are arranged in a radial manner, the size and number of the bipolar electrodes are greatly limited due to the influence of the circumferential angle and the radiation radius. The multiple closed bipolar electrodes arranged in series and parallel combination are not affected by the factors.
The aim of the invention is achieved by the following technical scheme:
an electrochemiluminescence chip comprises an integrated bipolar electrode, a driving electrode anode and a driving electrode cathode, a supporting channel and a reporting channel;
one end of the integrated bipolar electrode is a bipolar electrode cathode;
the other end of the integrated bipolar electrode is divided into a plurality of parallel branches; the parallel branches cross more than one reporting channel; the part of the parallel branch which is arranged in the report channel is the bipolar electrode anode, and the bipolar electrode anode on each parallel branch is in series connection; thus, the bipolar electrode anodes of the chip have a series connection relationship and a parallel connection relationship;
further, the report channel may only contain one bipolar electrode anode, or may contain a plurality of bipolar electrode anodes on different parallel branches;
the report channel also contains a driving electrode negative electrode;
the support channel contains a bipolar electrode cathode and a driving electrode anode;
the integrated bipolar electrode can be a solid integrated bipolar electrode made of conductive materials, or can be formed by connecting bipolar electrode cathodes and anodes by conductors;
preferably, the parallel branches are distributed in parallel; the parallel distribution can keep the anode size of the bipolar electrode consistent, which is beneficial to detection;
further preferably, the parallel branches are distributed in parallel and are symmetrically distributed along the central axis of the integrated bipolar electrode where the bipolar electrode cathode is located;
the driving electrode cathode can be an integrated driving electrode cathode which spans a plurality of reporting channels and supplies power to the reporting channels simultaneously; the negative electrode of the driving electrode can also be respectively arranged in each report channel to supply power individually according to the requirement;
preferably, the bipolar electrode cathode modifies the nanomaterial to facilitate electron transfer and provide a high specific surface area, thereby improving electrochemiluminescence intensity and detection sensitivity;
the nano material is a multiwall carbon nano tube, gold nano particles or graphene quantum dots in the prior art;
preferably, before the multiwall carbon nanotubes are modified, chitosan is firstly dripped on the cathode of the bipolar electrode;
further, the chitosan and the multi-wall carbon nano tube are respectively dissolved in acetic acid and phosphate buffer solution;
the bipolar electrode anode width is 500-800 μm, preferably 600 μm;
the bipolar electrode cathode width is 700-1000 μm, preferably 800 μm;
the substrate of the chip can be cloth, paper, conductive glass or Polydimethylsiloxane (PDMS) and the like.
The electrochemical light emitting chips of the present invention can be manufactured using prior art methods, such as those described in literature (Sensors and Actuators B,2018, 270:341-352), literature (Biosensors and Bioelectronics,2019, 130:55-64), except for the shape and relative positions of the channels and electrodes, and the substrate material, etc.
A sensor comprises the electrochemiluminescence chip.
The electrochemiluminescence chip and the sensor can be used for unit detection sensing of characteristic components or simultaneous multiplex detection sensing;
the characteristic components comprise hydrogen peroxide, uric acid, glucose, lactic acid, choline, cholesterol and the like;
the electrochemical luminescence chip is used for detecting and sensing hydrogen peroxide and comprises the following steps:
electrolyte buffer solution is added into the supporting channel, and to-be-detected solution containing luminol, hydrogen peroxide and the electrolyte buffer solution is added into the reporting channel; applying voltage to the driving electrode, and exciting electrochemical luminescence reaction of luminol and hydrogen peroxide on the anode of the bipolar electrode to realize detection and sensing;
the electrochemical luminescence chip is used for detecting and sensing uric acid and comprises the following steps:
selecting a bipolar electrode cathode of the modified multiwall carbon nanotube, and modifying urate oxidase on the bipolar electrode anode; adding electrolyte buffer solution into the support channel, adding to-be-detected solution containing luminol and the electrolyte buffer solution into the report channel, and incubating for a period of time to enable enzyme catalytic reaction to fully proceed; fixing the chip on the bracket, and connecting the driving electrode with an external direct current power supply by using a lead; then the chip is moved into a camera, the anode of the bipolar electrode is arranged in the visual field of the camera, and the focal length is adjusted to enable the visual field to be clear; then starting a CCD imaging acquisition function, and immediately switching on an external power supply to apply a driving voltage to the bipolar electrode; the multi-wall carbon nano tube modified on the bipolar electrode cathode provides a high specific surface area, promotes electron transfer, and reduces dissolved oxygen molecules into water molecules; uric acid generates hydrogen peroxide under the action of urate oxidase, luminol becomes an excited state under the action of hydrogen peroxide and driving voltage, and then generates blue-violet light with the wavelength of 425nm in the process of returning the excited state to a ground state, so that detection and sensing are realized;
the electrochemical luminescence chip is used for simultaneously and multiplex detecting and sensing uric acid and glucose, and comprises the following steps:
modifying urate oxidase and glucose oxidase on bipolar electrode anode respectively, adding electrolyte buffer solution into support channel, and adding solution to be tested containing luminol and electrolyte buffer solution into report channel; uric acid and glucose respectively generate hydrogen peroxide under the action of two oxidases, when a driving voltage is applied to a driving electrode, the electrochemical luminescence reaction of luminol and hydrogen peroxide on an anode of the bipolar electrode is simultaneously excited, so that simultaneous multi-element detection and sensing are realized;
the electrolyte buffer is carbonate buffer or phosphate buffer, preferably carbonate buffer;
the pH value of carbonate buffer solution in the report channel and the support channel is 9.5-11.0, preferably 10.0 or 10.5;
the driving voltage is 8-12V, preferably 10V;
the luminol concentration is 1.5-2.5mM, preferably 2mM;
the urate oxidase concentration is 0.08-0.14U/. Mu.L, preferably 0.1U/. Mu.L;
the incubation time is 2-3.5min, preferably 2.5min.
Compared with the prior art, the invention has the following advantages and effects:
1. the invention provides a microfluidic chip with serially connected bipolar electrode anodes, which is connected with the same bipolar electrode cathode for the first time. This reduces the bipolar electrode cathode modification process and allows detection of multiple targets of different sample sources. In terms of the current flowing through the closed bipolar electrode, the magnitude of the current passing through each bipolar electrode anode is essentially the same, which is more advantageous for examining the lighting conditions of different areas to evaluate the accuracy of the detection.
2. Compared with the reported microfluidic chip unit detection technology, the multiple serial bipolar electrode anodes are parallel and symmetrically distributed along the electrode part between the bipolar electrode cathode and the anode, the closed bipolar electrode comprises one cathode and multiple serial and parallel anodes, multiple results can be generated at one time of detection, and the method has the advantages of less sample consumption, high detection efficiency and high detection accuracy.
3. The invention provides a novel power supply mode, namely, different driving voltages are simultaneously or individually supplied so as to perform electrochemical luminescence detection of a plurality of series-parallel closed bipolar electrodes, thereby meeting the requirements of simultaneous or non-simultaneous multi-element and accurate detection.
4. The number and the size of the bipolar electrode anodes arranged in series and parallel can be increased or decreased according to the requirements, and the bipolar electrode anodes are not influenced by the circumferential angle and the radius of the circle.
5. The accurate detection of the invention is not only suitable for anode sensing, but also suitable for cathode sensing.
6. The invention modifies the multiwall carbon nanotube at the cathode of the bipolar electrode, which can increase the transfer rate of electrons and improve the detection sensitivity.
7. The invention only needs about 2.5min from sample adding to analysis completion, has simple operation and can realize rapid and quantitative detection.
8. The invention can meet the requirements of simultaneous or non-simultaneous accurate multi-element detection (including multi-element detection of the same sample or different samples), and has extremely important significance in the fields of environmental monitoring, food safety detection, disease diagnosis and the like.
Drawings
FIG. 1 is a schematic diagram of the structure of a chip of the present invention;
FIG. 2 is a schematic diagram of the structure of the chip of the present invention;
wherein, 1-12: bipolar electrode anode, 13: bipolar electrode cathode, 14: drive electrode positive electrode, 15: driving electrode negative electrode, 16: support channels, 17-20: report channel, 21: a wax dam.
FIG. 3 is a typical imaging plot of electrochemiluminescence intensity values versus hydrogen peroxide concentration in chip A.
FIG. 4 is a graph of the electrochemiluminescence intensity value versus hydrogen peroxide concentration in chip A.
FIG. 5 is a schematic diagram of the structure of the chip of the present invention; wherein, 1-12: bipolar electrode anode, 13: bipolar electrode cathode, 14: drive electrode positive electrode, 15: driving electrode negative electrode, 16: support channels, 17-20: report channel, 21: a wax dam.
FIG. 6 is a representative image of electrochemiluminescence intensity values versus hydrogen peroxide concentration in chip B.
FIG. 7 is a graph of the electrochemiluminescence intensity value versus hydrogen peroxide concentration in chip B.
FIG. 8 is a schematic diagram of the structure of the chip of the present invention; wherein, 1-6: bipolar electrode anode, 13: bipolar electrode cathode, 14: drive electrode positive electrode, 15: driving electrode negative electrode, 16: support channels, 17-18: report channel, 21: a wax dam.
FIG. 9 is a graph of electrochemiluminescence intensity values on chip C versus uric acid and glucose at various concentrations.
Fig. 10 is a schematic structural diagram of a chip D; wherein, 4-6: bipolar electrode anode, 13: bipolar electrode cathode, 14: drive electrode positive electrode, 15: driving electrode negative electrode, 16: support channel, 18: report channel, 21: a wax dam.
FIG. 11 is a graph showing the relationship between the electrochemical luminescence intensity value and uric acid concentration in the chip D.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto.
Example 1
The multi-channel closed bipolar electrochemical luminescence chip (referred to as chip a in this embodiment) is manufactured and tested as follows:
1. the configuration of the chip A was designed using drawing software Adobe Illustrator CS, the carbon electrodes (drive electrode and closed bipolar electrode) were made by carbon screen printing techniques, and the support channels and report channels were made by solid wax screen printing techniques.
The obtained chip is shown in figures 1, 2, 5 and 8, and comprises an integrated bipolar electrode, a driving electrode positive 14 and a driving electrode negative 15, a supporting channel 16 and reporting channels 17-20;
one end of the integrated bipolar electrode is a bipolar electrode cathode 13;
the other end of the integrated bipolar electrode is divided into six parallel branches; each parallel branch spans two reporting channels; the parts of the parallel branches which are arranged in the report channels 17-20 are bipolar electrode anodes 1-12;
the report channel 17-20 also contains a driving electrode negative electrode 15;
the support channel 16 contains a bipolar electrode cathode 13 and a driving electrode anode 14;
further, the report channel may contain multiple bipolar electrode anodes (fig. 1, 5 and 8) on different parallel branches, or may contain only one bipolar electrode anode (fig. 2);
2. by adopting a luminol-hydrogen peroxide system, a plurality of experimental groups are arranged to examine the electrochemiluminescence intensity of the chip A when hydrogen peroxide with the same concentration is accurately detected at the same time. The hydrogen peroxide concentrations were set to 0mM,0.1mM,0.5mM,2.5mM, respectively.
Fixing a cloth chip on a plastic bracket, adhering a conductive adhesive tape on a driving electrode, and connecting the driving electrode with an external direct current power supply (model LW-6403 KDS) by using a wire; adding carbonate buffer (pH 8) to the support channel, and adding a test solution (pH 8) comprising 2mM luminol, hydrogen peroxide and carbonate buffer to the reporter channels 17-20; moving the cloth chip into a camera bellows, placing the bipolar electrode anode in the field of view of a CCD camera (model MC 15), and adjusting the focal length to make the field of view clear; then the CCD camera imaging acquisition function is started, the photographing time interval is set to be 0.01s, a power supply is switched on, 10V driving voltage is applied to the bipolar electrode, the electrochemiluminescence reaction is triggered, and the PC stores pictures of electrochemiluminescence signals. The imaging data were further analyzed by Matlab R2015a (MathWorks company, USA) and Origin 8.5 (Microcal Software inc., newark, USA) software.
The electrochemiluminescence typical imaging diagram is shown in fig. 3, and the relationship between the electrochemiluminescence intensity value and the hydrogen peroxide concentration is shown in fig. 4.
As can be seen from fig. 4, the electrochemiluminescence intensities on the 12 bipolar electrode anodes are substantially uniform with a relative standard deviation of no more than 5%. Therefore, the chip A has good accurate detection potential.
Example 2
The multi-path closed bipolar electrochemical luminescence chip (called as a chip B in the embodiment) is characterized in that the driving electrode negative electrode is divided on the basis of the cloth chip A, and the manufacturing and detection processes are similar to the chip A.
1. FIG. 5 is a schematic diagram of the structure of a cloth chip B, which contains bipolar electrode anodes 1-12, bipolar electrode cathodes 13, driving electrode anodes 14, driving electrode cathodes 15, support channels 16 and reporting channels 17-20.
The chip a cannot supply power to the bipolar electrode anode in each report channel separately during the test, and in order to provide a more diversified test mode for the chip, it is necessary to supply power to the driving electrodes individually. The negative electrode of one driving electrode of the chip A is divided into four, and driving voltages can be respectively provided for the bipolar electrode anodes in the four reporting channels.
2. Several experimental groups are set to examine the application of the chip B in detecting hydrogen peroxide with different concentrations in an individual power supply mode. To the reporter channels 17-20, test solutions (pH 8) containing 2mM luminol, hydrogen peroxide at different concentrations (0 mM,0.1mM,0.5mM,2.5 mM) and carbonate buffer were added, respectively, and to the support channel 16, carbonate buffer (pH 8) was added. A 10V drive voltage is supplied to the drive electrodes to cause the electrochemical luminescence reactions to occur sequentially at the bipolar electrode anodes located in the reporting channels 17-20.
The typical imaging diagram of electrochemiluminescence is shown in fig. 6, and the relationship between the intensity value of electrochemiluminescence and the concentration of hydrogen peroxide is shown in fig. 7.
As can be seen from fig. 6 and 7, the electrochemiluminescence reaction at the bipolar electrode anode sequentially occurs with the sequential supply of the driving voltage. Therefore, the chip B has good non-simultaneous accurate detection capability.
Example 3
The multi-path closed bipolar electrochemical luminescence chip (called chip C in the embodiment) simplifies the number of bipolar electrode anodes and reporting channels on the basis of a cloth chip A, and the manufacturing and detection process is similar to that of the chip A.
1. Fig. 8 is a schematic diagram of the structure of the chip C, which contains bipolar electrode anodes 1 to 6, bipolar electrode cathode 13, driving electrode anode 14, driving electrode cathode 15, support channel 16 and reporting channels 17, 18.
2. Several experimental groups were set up to examine chip C for simultaneous, accurate detection of uric acid and glucose, with uric acid and glucose concentrations set up at several different concentrations (0 mM,0.1mM, 0.25 mM).
Modifying 5U/. Mu.L glucose oxidase on the bipolar electrode anode 1-3, modifying 0.1U/. Mu.L urate oxidase on the bipolar electrode anode 4-6, pre-fixing chitosan and multi-wall carbon nano-tubes on the bipolar electrode cathode 13; to each of the report channels 17, 18, 20. Mu.L of a mixed test solution (pH 10) containing luminol, carbonate buffer, uric acid and glucose was added, and to the support channel 16, 10. Mu.L of carbonate buffer (pH 10.5) was added. Incubation is carried out for a period of time sufficient for the enzyme-catalyzed reaction to proceed. Moving the cloth chip into a camera bellows, placing the bipolar electrode anode in the field of view of a CCD camera (model MC 15), and adjusting the focal length to make the field of view clear; the automatic photographing function of the CCD camera is started, the photographing time interval is set to be 0.01s, and meanwhile, the power supply is turned on, so that the electrochemical luminescence reaction occurs at the same time on the bipolar electrode anodes in the reporting channels 17 and 18. The PC will save a picture of the electrochemiluminescence signal. The imaging data were further analyzed by Matlab R2015a (MathWorks company, USA) and Origin 8.5 (Microcal Software inc., newark, USA) software.
The detection results are shown in FIG. 9. From the experimental results, it can be seen that: the bipolar electrode anodes in the reporting channels 17 and 18 can be independently detected without mutual influence, so that the chip C can be used for accurately detecting uric acid and glucose at the same time.
Comparative example
The multi-channel closed bipolar electrochemical luminescence chip (referred to as chip D in this comparative example) is simplified from chip C, and the manufacturing and detection process is similar to that of chip C.
1. Fig. 10 is a schematic diagram of the structure of the cloth chip D including bipolar electrode anodes 4 to 6, bipolar electrode cathode 13, driving electrode anode 14, driving electrode cathode 15, support channel 16 and report channel 18.
2. The bipolar electrode cathode of the chip D is pre-fixed with chitosan and multiwall carbon nanotubes, and the anode is pre-fixed with urate oxidase, and the specific process is as follows:
firstly, dripping chitosan (2.5 mg/mL) on the bipolar electrode cathode 13, and drying at room temperature for about 10 minutes; then, the multi-wall carbon nano tube (5 mg/mL) is dripped on the cathode, and the cathode is dried for about 10 minutes at room temperature; dripping urate oxidase on the bipolar electrode anodes 4-6, and drying at room temperature for about 10 min; after the pre-fixing is completed, the cloth chip is placed in a refrigerator at 4 ℃ in a sealing manner.
To the support channel 16 10. Mu.L of carbonate buffer was added and to the reporter channel 18 20. Mu.L of the test solution comprising luminol and carbonate buffer was added and incubated for a period of time sufficient for the enzyme-catalyzed reaction to proceed. Moving the cloth chip into a camera bellows, placing the bipolar electrode anode in the field of view of a CCD camera (model MC 15), and adjusting the focal length to make the field of view clear; starting an automatic photographing function of the CCD camera, setting a photographing time interval to be 0.01s, simultaneously switching on a power supply to trigger an electrochemiluminescence reaction, and storing pictures of electrochemiluminescence signals by the PC. The imaging data were further analyzed by Matlab R2015a (MathWorks company, USA) and Origin 8.5 (Microcal Software inc., newark, USA) software.
3. Several important factors affecting the electrochemiluminescence intensity in the comparative example (bipolar electrode anode and cathode width, driving voltage, luminol concentration, urate oxidase concentration, incubation time, reporter channel carbonate buffer pH and support channel carbonate buffer pH) were preferred, with corresponding optimized values of 600 μm, 800 μm, 10V, 2mM, 0.1U/μl, 2.5min, 10 and 10.5, respectively.
4. The dynamic curve for quantitative detection of uric acid on the bipolar electrode anode 4-6 of chip E is shown in FIG. 11. The electrochemiluminescence signal intensity increases with increasing uric acid concentration. The electrochemiluminescence intensity (represented by Y) and the uric acid concentration (represented by X) are in a linear relation within a certain range (0.01-0.1 mM, 0.25-1 mM); for bipolar electrode anode 4 (BPE 4), its linear equation may be expressed as Y BPE4 ＝6.875X-0.023(R 2 =0.9980) and Y BPE4 ＝0.671X+0.955(R 2 =0.9814); for bipolar electrode anode 5 (BPE 5), its linear equation may be expressed as Y BPE5 ＝6.763X-0.023(R 2 = 0.9981) and Y BPE5 ＝0.760X+0.907(R 2 = 0.9877); for bipolar electrode anode 6 (BPE 6), its linear equation may be expressed as Y BPE6 ＝6.743X-0.022(R 2 =0.9985) and Y BPE6 ＝0.697X+0.943(R 2 = 0.9901). The calculation method for the detection limit comprises the following steps: xl=xb+3sb, where Xb is the signal intensity at the time of the blank control, sb is the standard deviation of the blank control5 replicates). The detection limits of uric acid corresponding to BPE4, BPE5 and BPE6 are 5.2 mu M, 5.3 mu M and 4.9 mu M respectively calculated by the obtained XL values. The relative standard deviation of the three detection limits is less than 3%, so that the method has potential for accurately detecting uric acid.
At the two uric acid concentrations set in example 3, there was no statistical difference in the electrochemiluminescence intensity value obtained by chip C compared with the electrochemiluminescence intensity value obtained by chip D in the comparative example. The chip C described in example 3 can be used for simultaneously and precisely detecting uric acid and glucose. In addition, the chip described in the comparative example can only be used for multi-target detection of the same sample source, while the chip described in the examples can satisfy simultaneous or non-simultaneous accurate detection of multiple targets of different sample sources.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (6)
1. An electrochemiluminescence chip comprises an integrated bipolar electrode, a driving electrode anode and a driving electrode cathode, a supporting channel and a reporting channel; the method is characterized in that:
one end of the integrated bipolar electrode is a bipolar electrode cathode;
the other end of the integrated bipolar electrode is divided into a plurality of parallel branches;
the parallel branches cross more than one reporting channel;
the part of the parallel branch which is arranged in the report channel is the bipolar electrode anode;
the report channel contains a bipolar electrode anode or contains a plurality of bipolar electrode anodes on different parallel branches;
the driving electrode negative electrode is an integrated driving electrode negative electrode and spans across a plurality of reporting channels; or the cathodes of the driving electrodes are respectively arranged in each report channel; the parallel branches are distributed in parallel and are symmetrically distributed along the central axis of the integrated bipolar electrode where the bipolar electrode cathode is located.
2. The electrochemical luminescence chip of claim 1, wherein: the bipolar electrode cathode modifies the nanomaterial.
3. The electrochemical luminescence chip of claim 1, wherein:
the report channel also contains a driving electrode negative electrode;
the support channel contains bipolar electrode cathode and driving electrode anode.
4. A sensor comprising the electrochemical luminescence chip of any of claims 1-3.
5. Use of an electrochemical luminescence chip according to any of claims 1-3 or a sensor according to claim 4 for unit detection sensing or simultaneous multiplex detection sensing of characteristic components.
6. The use according to claim 5, characterized in that: the characteristic components comprise hydrogen peroxide, uric acid, glucose, lactic acid, choline and cholesterol.
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| CN103969305A (en) * | 2014-04-30 | 2014-08-06 | 陕西师范大学 | Bipolar-electrode electrochemiluminescence imaging electrolytic cell |
| CN108303538A (en) * | 2018-01-26 | 2018-07-20 | 南京大学 | Two-way electrochemical luminescence colour developing switchs the application in row gland cancer multi-tracer before detection |
| CN110412021A (en) * | 2019-08-06 | 2019-11-05 | 华南师范大学 | A kind of electrochemical luminescence micro-fluidic chip of shared bipolar electrode cathode and its application |
| CN112362708A (en) * | 2020-10-19 | 2021-02-12 | 济南大学 | Preparation method of self-powered bipolar microelectrode microfluidic chip photoelectrochemical aptamer sensor |
| CN113884481A (en) * | 2021-09-29 | 2022-01-04 | 华南师范大学 | Dry bipolar electrochemical luminescence chip and application thereof in immunodetection |
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| CN103969305A (en) * | 2014-04-30 | 2014-08-06 | 陕西师范大学 | Bipolar-electrode electrochemiluminescence imaging electrolytic cell |
| CN108303538A (en) * | 2018-01-26 | 2018-07-20 | 南京大学 | Two-way electrochemical luminescence colour developing switchs the application in row gland cancer multi-tracer before detection |
| CN110412021A (en) * | 2019-08-06 | 2019-11-05 | 华南师范大学 | A kind of electrochemical luminescence micro-fluidic chip of shared bipolar electrode cathode and its application |
| CN112362708A (en) * | 2020-10-19 | 2021-02-12 | 济南大学 | Preparation method of self-powered bipolar microelectrode microfluidic chip photoelectrochemical aptamer sensor |
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