CN113219022A - Multi-channel digital microfluidic detection platform and application thereof - Google Patents
Multi-channel digital microfluidic detection platform and application thereof Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 114
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 claims abstract description 33
- 229960005091 chloramphenicol Drugs 0.000 claims abstract description 33
- 238000000835 electrochemical detection Methods 0.000 claims abstract description 16
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- 238000000034 method Methods 0.000 claims description 28
- 239000007788 liquid Substances 0.000 claims description 16
- 239000010931 gold Substances 0.000 claims description 7
- 229910052737 gold Inorganic materials 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 238000001903 differential pulse voltammetry Methods 0.000 claims description 4
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- 238000002484 cyclic voltammetry Methods 0.000 claims description 3
- 238000001179 sorption measurement Methods 0.000 claims description 3
- 238000004365 square wave voltammetry Methods 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 238000005516 engineering process Methods 0.000 abstract description 8
- 239000000243 solution Substances 0.000 description 10
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Abstract
本发明公开了一种多通道数字微流控检测平台及其应用，主要由检测电极芯片、底层和顶盖组成，底层和顶盖通过绝缘手段固定连接且保持一定距离；底层从下到上依次为底层绝缘层、底层电极层、底层介电层和底层疏水层；顶盖从下到上依次为顶盖疏水层和顶盖电极层。底层电极层和顶盖电极层中布置有电极片，上下一一对应；在顶盖上开有凹槽，用于放置检测电极芯片；检测电极芯片为丝网印刷电极芯片。本发明将多通道数字微流控技术与氯霉素电化学检测相结合，通过单接口实现多通道的微流控装置检测氯霉素，并且丝网印刷电极芯片的体积小，能够节约检测成本，实现快速、连续和便捷的在线检测，且多通道微流控平台并不局限于检测氯霉素。
The invention discloses a multi-channel digital microfluidic detection platform and its application. It mainly consists of a detection electrode chip, a bottom layer and a top cover. The bottom layer and the top cover are fixedly connected by insulating means and keep a certain distance; the bottom layer is sequentially from bottom to top It is the bottom insulating layer, the bottom electrode layer, the bottom dielectric layer and the bottom hydrophobic layer; the top cover is the top cover hydrophobic layer and the top cover electrode layer from bottom to top. Electrode sheets are arranged in the bottom electrode layer and the top cover electrode layer, and the top and bottom correspond to each other; grooves are opened on the top cover for placing detection electrode chips; the detection electrode chips are screen-printed electrode chips. The invention combines the multi-channel digital microfluidic technology with the electrochemical detection of chloramphenicol, realizes the detection of chloramphenicol by a multi-channel microfluidic device through a single interface, and the screen-printed electrode chip is small in size, which can save the detection cost , to achieve fast, continuous and convenient online detection, and the multi-channel microfluidic platform is not limited to the detection of chloramphenicol.
Description
Technical Field
The invention belongs to the field of micro analysis, and particularly relates to a multi-channel digital microfluidic detection platform and application thereof.
Background
Microfluidic chips have many advantages over traditional laboratories as carriers for experiments: the consumption of the sample and the reaction reagent is reduced, so that the cost is saved, the reaction time can be reduced, and the efficiency is improved; the parallel processing can be carried out to improve the experimental flux, and a new method is provided for the fields of screening of medicine preparation and system testing; the portable nature of the chip makes on-site medical diagnosis and bioanalysis feasible. With the development of the MEMS technology, the digital microfluidic chip has been a breakthrough in the processing technology and the driving and controlling technology of the micro-droplets, and has been widely applied in the fields of biology, chemistry, medicine, etc. depending on its own advantages. From the wide application in various fields, as a new technology, the digital microfluidic chip has the advantages of small volume, small reagent usage amount, fast reaction, easy carrying, parallel processing, easy realization of automation and the like, and in view of the unique advantages and the use value, more and more institutions and scholars are put into the microfluidic research team to seek the substantial breakthrough of the microfluidic in various fields. The improvement of the reliability, the efficiency and the stability of the experiment becomes the main content of research of people, and the development of the MEMS high-precision processing technology also provides guarantee for the miniaturization, the automation and the integration of the digital microfluidic chip.
As one of microfluidics, a digital microfluidic chip based on dielectric wetting generally consists of a solid substrate, a microelectrode deposited on the substrate, a dielectric layer coated on the surface of the micromotor and a hydrophobic layer on the uppermost layer, and realizes basic operations such as generation, movement, splitting, mixing and the like of liquid drops by applying voltages to the electrodes in a certain sequence. Compared with other methods, the method is more flexible, a plurality of micro-droplets can be controlled simultaneously by controlling the voltage-applying time sequence, no influence is caused between the micro-droplets, and the efficiency is higher; when part of the electrodes are damaged, the electrode can be continuously used without influencing other units to generate liquid drops; has wide application range and is almost suitable for any solution.
At present, the commonly used electrochemical detection means are based on a three-electrode system, mainly comprise a reference electrode, an auxiliary electrode (counter electrode) and a working electrode, and in most laboratories at present, three electrode bars are used, and have the defects of large volume, inconvenient replacement after use and the like. The silk-screen printing electrode is a shaped and solidified electrode prepared by adopting a silk-screen printing technology to print ink on an inert solid plane substrate (such as PVC, glass fiber, ceramic, polyester film, aluminum oxide, paper and the like) through layer-by-layer deposition, forming an electrode pattern by utilizing a silk screen, and then baking the electrode to remove a solvent in the ink. The screen printing electrode has the characteristics of flexible design, easy batch production, strong stability, easy miniaturization, low cost and the like. However, in order to place the electrodes on the hardware circuit conveniently, the pins of the current screen-printed electrodes are generally left long, which causes great waste and cost increase for batch detection, so that the problem of long protruding pins of the current screen-printed electrodes is urgently needed to be improved.
While the chloramphenicol can treat various infections, the chloramphenicol residue has serious harm to human health. The traditional chloramphenicol detecting instrument has the defects of high cost, large volume, complex operation, high professional requirement and the like. Meanwhile, most of chloramphenicol detection methods are suitable for food or water quality detection, and multi-channel detection is needed due to the fact that a large number of samples are used, and existing solutions mostly complete multi-channel detection of multiple samples on the basis of sacrificing the circuit size of hardware, so that a large number of three-electrode screen printing electrodes are needed to be used at the same time, and cost is increased.
Disclosure of Invention
The invention aims to provide a multi-channel digital microfluidic detection platform and application thereof aiming at the defects of the prior art.
The purpose of the invention is realized by the following technical scheme: a multi-channel digital micro-fluidic detection platform is characterized by mainly comprising a detection electrode chip, a bottom layer and a top cover, wherein the bottom layer and the top cover are fixedly connected through an insulating means and keep a certain distance. The bottom layer comprises a bottom insulating layer, a bottom electrode layer, a bottom dielectric layer and a bottom hydrophobic layer from bottom to top in sequence. Electrode plates are arranged in the bottom electrode layer, and detection of different channels is realized by arrangement of different electrode plates; the edge electrode pads in the bottom electrode layer are the inlets of the channels. The top cover is a top cover hydrophobic layer and a top cover electrode layer from bottom to top in sequence. The arrangement of the electrode plates in the top cover electrode layer is in one-to-one correspondence with the electrode plates of each channel of the bottom electrode layer from top to bottom to form a complete loop. The top cover is provided with a groove for placing a detection electrode chip. The detection electrode chip is a screen printing electrode chip, and pins of the detection electrode chip are arranged according to electrode plates in the top cover electrode layer. The position of the detection electrode chip is a detection area.
Further, the working process specifically comprises: modifying a working electrode of the detection electrode chip; then, placing the modified detection electrode chip at the groove of the top cover, dripping the liquid drops to be detected on different channels, and detecting the concentration of the liquid drops to be detected by adopting an electrochemical detection method; during detection, the liquid drops to be detected of different channels sequentially move to a detection area for detection by applying electric potential on the electrode sheets of the channels, so that continuous detection is realized.
Further, the electrochemical detection method is cyclic voltammetry, time current method, differential pulse voltammetry, square wave voltammetry or other electrochemical detection means.
Further, the working electrode of the modified detection electrode chip is specifically: the solution of the modified electrode with a certain concentration is placed on a channel, electric potential is applied to an electrode sheet of the channel to enable the modified electrode to move to a detection area, and nanoparticles and other modified substances are deposited on a working electrode of a detection electrode chip 7 by adopting an electrochemical deposition method or a physical adsorption method.
Furthermore, the pattern of the electrode plate is cross-shaped or meter-shaped; the electrode pads on each channel are wired to achieve control of each electrode pad.
Further, the empty channels in the bottom electrode layer may be used to store waste fluids, cleaning fluids, solutions for modifying the electrodes, and other reagents or solutions for other purposes.
Furthermore, the groove on the top cover is in a circular truncated cone shape or a quadrangular truncated cone shape and is used for placing a circular or square detection electrode chip.
Furthermore, the surface of the top cover groove is provided with a notch which is convenient for dismounting and mounting the detection electrode chip.
Furthermore, round holes convenient for dripping the liquid drops to be detected are reserved at inlets of all channels of the top cover.
A method for detecting chloramphenicol concentration based on a multi-channel digital microfluidic detection platform specifically comprises the following steps: and dripping chloroauric acid solution in the detection area to ensure that the nano gold is deposited on the surface of the working electrode of the detection electrode chip. And (3) placing the detection electrode chip modified by the nanogold at the groove of the top cover, then dropwise adding the chloramphenicol sample to be detected on different channels, and detecting the concentration of the chloramphenicol sample to be detected by adopting an electrochemical detection method.
The invention has the beneficial effects that:
1. the invention combines the multichannel digital microfluidic technology with the electrochemical detection of chloramphenicol, realizes the multichannel microfluidic device to detect the chloramphenicol through a single interface, requires the volume of a screen printing electrode chip to be as small as possible, can save the detection cost, realizes the rapid, continuous and convenient online detection, and is not limited to the detection of the chloramphenicol by a multichannel microfluidic platform.
2. The multi-channel digital micro-fluidic platform can rapidly and immediately detect the chloramphenicol residues in food, and compared with the traditional hardware circuit for realizing multi-channel electrochemical detection, the multi-channel digital micro-fluidic platform can save the volume of the hardware circuit under the condition of realizing multi-channel, thereby achieving the portable effect. Moreover, the design of the groove above the multi-channel micro-fluidic platform can overcome the defect that the screen printing extends out of the electrode for a long time.
Drawings
FIG. 1 is a schematic structural diagram of a multi-channel digital microfluidic detection platform;
FIG. 2 is a top view of a bottom layer of the multi-channel digital microfluidic detection platform;
FIG. 3 is a top view and a bottom view of the multi-channel digital microfluidic detection platform in a top cover detection state;
FIG. 4 is a diagram of a groove on the top cover of the multi-channel digital microfluidic detection platform and a corresponding screen printed electrode chip;
FIG. 5 is a schematic diagram of electrochemical detection of chloramphenicol;
FIG. 6 is a graph showing the results of the detection of chloramphenicol samples at different concentrations;
in the figure: the device comprises a bottom insulating layer 1, a bottom electrode layer 2, a bottom dielectric layer 3, a bottom hydrophobic layer 4, a top electrode layer 7, a top cover hydrophobic layer 6, an insulating support 5, a liquid drop 8 to be detected and a detection electrode chip 9.
Detailed Description
As shown in figure 1, the multi-channel digital microfluidic detection platform mainly comprises a detection electrode chip 9, a bottom layer and a top cover, wherein the bottom layer and the top cover are fixedly connected to complete positioning and keep a certain distance through an insulating support 5 or other insulating support means. Thus, even an inexperienced ordinary technician can complete the disassembly and assembly of the whole detection platform.
The bottom layer comprises a bottom insulating layer 1, a bottom electrode layer 2, a bottom dielectric layer 3 and a bottom hydrophobic layer 4 from bottom to top in sequence. The bottom layer insulating layer 1 is used for protecting the bottom layer electrode layer 2 and avoiding the interference of the outside on the bottom layer electrode layer 2; the bottom dielectric layer 3 and the bottom hydrophobic layer 4 are present to ensure that the multi-channel digital microfluidic detection platform can work normally.
As shown in fig. 2, the arrangement of different electrode sheets in the bottom electrode layer 2 can realize the detection of different channels, a plurality of channels are arranged in the bottom electrode layer 2, and each channel corresponds to different moving routes of the liquid drop 8 to be detected; the edge electrode pads in the bottom electrode layer 2 are the inlet of each channel; the pattern of the electrode arrangement can be designed into a cross shape, a meter shape and other shapes. Wiring the electrode plates on each channel to realize the control of each electrode plate; the wiring meets the control requirement and does not cause short circuit, and the movement of the liquid drops is controlled by adopting a conventional means after the wiring, for example, the movement of the liquid drops is controlled by using a singlechip. The empty channels in the bottom electrode layer 2 can be used to store waste fluids, cleaning fluids, solutions for modifying the electrodes, and reagents or solutions for other purposes.
The top cover is provided with a top cover hydrophobic layer 6 and a top cover electrode layer 7 from bottom to top in sequence. As shown in fig. 3, the arrangement of the electrode plates in the top cover electrode layer 7 is in one-to-one correspondence with the electrode plates of each channel of the bottom electrode layer 2, and the electrode plates on the top layer and the bottom layer are respectively connected with positive and negative potentials in the control circuit to form a complete loop. The top cover is provided with a circular truncated cone-shaped or quadrangular truncated cone-shaped groove, and the shape of the groove is shown in figure 4 and is used for placing a circular or square detection electrode chip 9. The surface of the top cover groove is provided with a notch which is convenient for dismounting and mounting the detection electrode chip 9. The top cover groove is used for keeping the detection electrode chip 9 and the electrode plate in the top cover electrode layer 7 in close contact, and meanwhile, round holes convenient for dripping liquid drops 8 to be detected are reserved at inlets of all channels of the top cover.
The detection electrode chip 9 is a screen printing electrode chip, and adopts two electrodes, three electrodes or four electrodes, and pins of the detection electrode chip need to be arranged by avoiding electrode plates in the top cover electrode layer 7, so that the pins on the top cover are prevented from being in contact with the electrode plates in the top cover electrode layer 7, and detected signals and electric potentials applied to the electrode plates in the top cover electrode layer 7 are mixed together to cause distortion of detection results. The position of the detection electrode chip 9 is a detection area.
The appearance shape, channel layout, and materials of each level of the present invention are not limited to the present specification.
The invention relates to a using process of a multi-channel digital microfluidic detection platform, which specifically comprises the following steps:
(1) modifying the working electrode of the detection electrode chip 9: placing a solution of a modified electrode with a certain concentration on a channel of a multi-channel digital microfluidic detection platform, applying potential on an electrode sheet of the channel to enable the modified electrode to move to a detection area, and depositing nano particles (nano gold) and other modified substances on a working electrode of a detection electrode chip 7 by adopting an electrochemical deposition method or a physical adsorption method; the working electrode modification can also be performed on the detection electrode chip 9 externally.
(2) The detection electrode chip 9 modified in the step (1) is reversely buckled and tightly attached to a groove of a top cover, then the liquid drop 8 to be detected is dripped on different channels, and the concentration of the liquid drop 8 to be detected is detected by adopting an electrochemical detection method; during detection, applying electric potential on the electrode plates of the channels to enable the liquid drops to be detected 8 of different channels to sequentially move to a detection area for detection, thereby realizing continuous detection; after the detection is finished, the detection electrode chip 9 on the top cover can be replaced according to the requirement. The electrochemical detection method is cyclic voltammetry, time current method, differential pulse voltammetry, square wave voltammetry or other electrochemical detection means.
Examples
In this embodiment, gold nano-deposition is performed on the surface of a working electrode of a three-electrode chip, and concentration detection is performed on a medium-concentration chloramphenicol test solution and a low-concentration chloramphenicol test solution, which specifically includes:
1g/L chloroauric acid solution is dripped into the detection area, and the nano-gold is deposited on the surface of the working electrode of the three-electrode chip by depositing for 1000s at a potential of-300 mV by using a time current method in electrochemical deposition. Chloramphenicol was formulated at concentrations of 0.01g/L, 0.02g/L, 0.05g/L, 0.1g/L, 0.2g/L, 0.5g/L, 1.0g/L using PBS buffer at pH 7.2-7.4. When a chloramphenicol sample is detected, a three-electrode system and the multi-channel digital microfluidic detection platform are used, a nanogold modified three-electrode chip is tightly attached to a groove of a top cover, chloramphenicol samples with different concentrations are then dripped on different channels, and the concentration of the chloramphenicol sample can be detected by adopting a differential pulse voltammetry method; in the process of detecting the chloramphenicol sample droplet of one channel, the chloramphenicol sample droplet of the next channel can slowly move towards the detection area, and the next chloramphenicol sample droplet immediately enters the detection area for detection after the previous chloramphenicol sample droplet leaves the detection area, so that continuous detection is realized.
The schematic diagram of chloramphenicol detection in this embodiment is shown in fig. 5, and since the working electrode of the screen-printed electrode is a carbon electrode, the electron conduction velocity of a common carbon electrode is relatively slow, and the generated electric signal is relatively weak, and meanwhile, studies show that gold nanoparticles have the advantages of enlarging the surface area of the electrode, increasing the electron conduction velocity, having high catalytic activity, and the like.
As shown in fig. 6, it can be seen that the peak current gradually increases as the concentration of the chloramphenicol sample droplet increases, the peak current and the concentration are analyzed, a strong linear relationship exists between the peak signal and the concentration, and for a certain droplet to be detected, the concentration of chloramphenicol in the droplet to be detected can be obtained according to the linear relationship between the measured peak current and the concentration, so as to implement the detection of the concentration of chloramphenicol. This detailed description is to be construed as illustrative only and is not to be taken in a limiting sense, as the invention is protected by the appended claims.
Claims (10)
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Citations (4)
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| CN102175744A (en) * | 2011-01-06 | 2011-09-07 | 复旦大学 | Electrochemical sensor chip with digital microfluidic technology |
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| CN109046429A (en) * | 2018-08-23 | 2018-12-21 | 青岛大学 | Mat gold nano particle/graphite phase carbon nitride composite material electrochemical sensor is detected for chloramphenicol |
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Patent Citations (4)
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| CN102175744A (en) * | 2011-01-06 | 2011-09-07 | 复旦大学 | Electrochemical sensor chip with digital microfluidic technology |
| CN103170383A (en) * | 2013-03-10 | 2013-06-26 | 复旦大学 | Nano-material electrode modification based electrochemical integrated digital micro-fluidic chip |
| CN109046429A (en) * | 2018-08-23 | 2018-12-21 | 青岛大学 | Mat gold nano particle/graphite phase carbon nitride composite material electrochemical sensor is detected for chloramphenicol |
| CN112034025A (en) * | 2019-06-04 | 2020-12-04 | 多助科技(武汉)有限公司 | Composite modified electrode for chloramphenicol detection and construction and detection methods thereof |
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