CN112473757A - Micro-fluidic chip detection system for food safety rapid detection - Google Patents
Micro-fluidic chip detection system for food safety rapid detection Download PDFInfo
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Abstract
本发明公开了一种用于食品安全快速检测的微流控芯片检测系统，涉及微流控芯片领域，包括：上位机、电化学工作站、继电器模块、磁场驱动模块和微流控芯片，上位机连接到继电器模块和电化学工作站，继电器模块连接到磁场驱动模块，磁场驱动模块连接到微流控芯片，微流控芯片连接到电化学工作站；微流控芯片包括上盖板和下基板，上盖板形成有微流通道，微流通道包括多个区域，分离区通过冲洗区连通到检测区，废液区连通到分离区和检测区，检测区中的三电极检测单元连接到电化学工作站，通过磁场驱动模块实现对待测样品的自动化流向控制，同时在不同区域之间设置可编程通断阀门使得各区域相互独立、互不干扰，更方便的对样品进行检测。
The invention discloses a microfluidic chip detection system for rapid detection of food safety, relates to the field of microfluidic chips, and includes: a host computer, an electrochemical workstation, a relay module, a magnetic field drive module and a microfluidic chip, the host computer Connected to the relay module and the electrochemical workstation, the relay module is connected to the magnetic field driving module, the magnetic field driving module is connected to the microfluidic chip, and the microfluidic chip is connected to the electrochemical workstation; the microfluidic chip includes an upper cover plate and a lower substrate, the upper The cover plate is formed with a microfluidic channel, the microfluidic channel includes a plurality of areas, the separation area is connected to the detection area through the flushing area, the waste liquid area is connected to the separation area and the detection area, and the three-electrode detection unit in the detection area is connected to the electrochemical workstation , The automatic flow direction control of the sample to be tested is realized through the magnetic field drive module, and programmable on-off valves are set between different areas to make each area independent and not interfere with each other, and it is more convenient to detect the sample.
Description
Technical Field
The invention relates to the field of microfluidic chips, in particular to a microfluidic chip detection system for food safety rapid detection.
Background
Food safety is a big matter related to national life, and is an international problem, so that the factors causing the food safety problem are very complex, and the quality detection can be carried out only by adopting a spot check mode when the food safety is monitored by food departments. At present, common food safety detection methods comprise a gas phase method, a liquid chromatography-mass spectrometry method and a spectroscopic analysis method, but the methods have certain defects, and are often operated in special laboratories, so that a large number of large-scale detection devices and a large number of detection reagents are often needed, the detection time is long, and the detection cost is high due to the fact that professional experimenters are needed for operation.
The micro-fluidic technology makes the appearance of a rapid and accurate detection means possible, the micro-fluidic technology has become a novel sensing platform, different analysis steps, biological identification materials and appropriate sensors can be ingeniously integrated on the platform, so that a new generation of sensors are generated for detecting mycotoxins, food-borne pathogens, medicines, allergens and the like, but the existing detection system has low degree of automation, cannot accurately control the flow direction of a sample to be detected in a chip, and has unsatisfactory detection effect and larger deviation from the actual detection.
Disclosure of Invention
The invention provides a micro-fluidic chip detection system for food safety rapid detection aiming at the problems and technical requirements, and the technical scheme of the invention is as follows:
a micro-fluidic chip detection system for food safety rapid detection comprises an upper computer, an electrochemical workstation, a relay module, a magnetic field driving module and a micro-fluidic chip, wherein the upper computer is connected to the relay module and the electrochemical workstation, the relay module is connected to the magnetic field driving module, the micro-fluidic chip is installed in the magnetic field driving module, and the micro-fluidic chip is connected to the electrochemical workstation;
the micro-fluidic chip comprises an upper cover plate and a lower cover plate, wherein the upper cover plate and the lower cover plate are bonded together, a micro-fluidic channel is formed in the upper cover plate and comprises a separation area, a washing area, a detection area and a waste liquid area, the separation area is communicated with the detection area through the washing area, the waste liquid area is communicated with the separation area and the detection area, the detection area comprises two detection channels and a three-electrode detection unit, the two detection channels are communicated with the washing area, one end of the three-electrode detection unit is connected to the two detection channels, the other end of the three-electrode detection unit is connected to the electrochemical workstation, and the electrochemical workstation determines the concentration of a sample to be detected in the micro-fluidic.
The separation area comprises a micro-reaction pool and a plurality of injection ports, a buffer solution injection port, an immunomagnetic bead injection port and a sample injection port to be detected are communicated with the micro-reaction pool, the waste liquid area comprises a first waste liquid pool and a second waste liquid pool, a first valve is arranged on a connecting channel between the first waste liquid pool and the micro-reaction pool, the flushing area comprises a cleaning pool and a cleaning liquid injection port, a second valve is arranged on a connecting channel between the micro-reaction pool and the cleaning pool, a third valve is arranged on a connecting channel between the cleaning pool and the detection area, and the flushing area is communicated with the second waste liquid pool through the two detection channels.
The technical scheme is that the three-electrode detection unit comprises three-electrode detection arrays, each three-electrode detection array comprises a reference electrode, two auxiliary electrodes and two working electrodes, each electrode comprises a connecting piece and a detection piece, the detection pieces are connected to the electrochemical workstation through the connecting pieces, the two detection pieces of one reference electrode are respectively positioned in the two detection channels, and each detection channel is also provided with the detection piece of one auxiliary electrode and the detection piece of one working electrode.
The connecting piece of the reference electrode is positioned between the two detection channels, the connecting piece of the first auxiliary electrode and the connecting piece of the first working electrode are positioned on one side of the two detection channels, and the connecting piece of the second auxiliary electrode and the connecting piece of the second working electrode are positioned on the other side of the two detection channels.
The further technical scheme is that the length of the detection piece of the reference electrode is greater than the sum of the length of the detection piece of the auxiliary electrode and the length of the detection piece of the working electrode, the transverse area of the detection piece of the auxiliary electrode and the transverse area of the detection piece of the working electrode are positioned in the transverse area of the detection piece of the reference electrode, and the transverse area of the detection piece of the auxiliary electrode and the transverse area of the detection piece of the working electrode are not overlapped.
The magnetic field driving module comprises eight electromagnetic coils, the relay module comprises eight relays, each electromagnetic coil is connected to one relay, and the upper computer controls the on-off of each relay.
The further technical scheme is that the determination of the concentration of the sample to be detected in the microfluidic chip comprises the following steps:
preparing n standard solutions with equal concentration gradients, wherein the concentration of each standard solution is known, enabling each standard solution to pass through a detection area, controlling the potential of a three-electrode detection unit to scan in a triangular waveform at different time rates, and establishing n current-potential curves of the standard solutions with different concentrations;
obtaining the signal intensity of each standard solution according to the current-potential curve, and establishing corresponding standard curves of different standard solution concentrations and signal intensity sizes through data fitting;
when a sample to be detected with unknown concentration is detected, the concentration of the sample to be detected can be obtained in the standard curve through the signal intensity obtained by the detection of the three-electrode detection unit.
The further technical scheme is that the expression of the concentration of the sample to be detected is as follows:
yt＝a+bxt；
wherein, ytRepresenting the signal intensity, x, of the sample to be testedtWhich is indicative of the concentration of the sample to be tested,
in the formula (I), the compound is shown in the specification,
wherein x isiDenotes the concentration of the i-th standard solution, yiIndicates the signal intensity of the i-th standard solution.
The beneficial technical effects of the invention are as follows: the automatic flow direction control of the sample to be detected is realized through the magnetic field driving module; meanwhile, different regions are arranged to facilitate the detection of the sample, so that the purpose that the regions do not interfere with each other is achieved; the three-electrode detection unit can acquire multiple groups of data, so that the detection result is more accurate, and the operation is simpler and more convenient.
Drawings
Fig. 1 is a schematic structural diagram of the detection system of the present application.
Fig. 2 is a top view of a microfluidic chip of the present application.
Fig. 3 is a perspective view of a microfluidic chip of the present application.
Fig. 4 is a perspective view of a three-electrode detection unit of the present application.
FIG. 5 is a top view of a three-electrode detection cell of the present application.
Fig. 6 is a schematic diagram of a magnetic field drive module of the present application.
Detailed Description
The following further describes the embodiments of the present invention with reference to the drawings.
As shown in fig. 1, a detection system for a microfluidic chip for rapid detection of food safety includes an upper computer, an electrochemical workstation, a relay module, a magnetic field driving module and a microfluidic chip, wherein the upper computer is connected to the relay module and the electrochemical workstation, the relay module is connected to the magnetic field driving module, the microfluidic chip is installed in the magnetic field driving module, the microfluidic chip is connected to the electrochemical workstation, the upper computer is used for controlling the whole detection system, and the electrochemical workstation is used for determining the concentration of a sample to be detected in the microfluidic chip.
As shown in fig. 2, the microfluidic chip includes an upper cover plate 1 and a lower cover plate 2, the upper cover plate is made of PDMS (polydimethylsiloxane), the lower cover plate is made of glass, the upper cover plate 1 and the lower cover plate 2 are bonded together, a microfluidic channel is formed on the upper cover plate, the microfluidic channel includes a separation area 3, a washing area 4, a detection area 5 and a waste liquid area 6, each area is communicated through a connecting channel, the separation area 3 is communicated to the detection area 5 through the washing area 4, the waste liquid area 6 is communicated to the separation area 3 and the detection area 5, and the detection area 5 is connected to an electrochemical workstation.
Preferably, the upper cover plate 1 and the lower base plate 2 are both rectangular parallelepiped shapes, the cross section of each connecting channel is rectangular, and the size of the upper cover plate 1 is preferably as follows: the length is 5cm, the width is 3cm, and the height is 4 mm; the dimensions of the lower substrate 2 are preferably: the length is 5.5cm, the width is 3.5cm, and the height is 1 mm; the cross-sectional dimensions of the connecting channel are preferably: the width is 0.2mm, and the length is 0.4 mm.
As shown in fig. 3, the separation region includes a micro-reaction cell 7 and a plurality of injection ports, the buffer solution injection port 8, the immunomagnetic bead injection port 9 and the sample injection port 10 to be tested are communicated with the micro-reaction cell, the waste liquid region 6 includes a first waste liquid cell 11 and a second waste liquid cell 12, a first valve 13 is disposed on a connection channel between the first waste liquid cell 11 and the micro-reaction cell 7, the washing region 4 includes a washing cell 14 and a washing liquid injection port 15, a second valve 16 is disposed on a connection channel between the micro-reaction cell 7 and the washing cell 14, a third valve 17 is disposed on a connection channel between the washing cell 14 and the detection region 5, the detection region 5 includes a three-electrode detection unit 18, a first detection channel 19 and a second detection channel 20, wherein the washing area 4 is communicated with the second waste liquid pool 12 through the two detection channels, one end of the three-electrode detection unit 18 is connected into the two detection channels, and the other end is connected to the electrochemical workstation.
When detection is performed, the first valve 13 is controlled to be opened, the second valve 16 is controlled to be closed, and the buffer solution is injected from the buffer solution injection port 8, so that the micro-reaction tank 7 and the first waste liquid tank 11 are filled with the buffer solution, and the buffer solution plays a role of a carrier and does not participate in actual reaction. Then, the first valve 13 is closed, immunomagnetic beads are injected from the immunomagnetic bead injection port 9, and a sample to be detected is injected from the sample injection port 10, and the injection operation of the injection port is completed by a multi-path constant pressure pump.
After the reaction in the micro-reaction tank 7 is completed, the sample to be tested is attached to the immunomagnetic beads, the third valve 17 is closed, the first valve 13 and the second valve 16 are opened, the cleaning solution is injected from the cleaning solution injection port 15, the cleaning solution flows to the first waste liquid tank 11 due to the acting force of the buffer solution, the immunomagnetic beads are driven by the magnetic field control module to flow from the micro-reaction tank 7 to the cleaning tank 14, the cleaning purpose of the cleaning solution is achieved, then the second valve 16 is closed, the third valve 17 is opened, and the sample to be tested flows to the detection area.
The three valves are all composed of metal solids, the appearance is a cylinder, the three valves completely penetrate through the upper cover plate and can move up and down simultaneously, the diameter of the cylinder is equal to the width of the connecting channel, the three valves respectively correspond to three independent valve controllers, the valves are controlled by the controllers to move up and down according to a detection flow, the on-off of fluid in the connecting channel is realized, and meanwhile, in order to ensure that the channel does not leak when the valves are closed, the surface of the channel needs to be plasticized.
As shown in fig. 4, the three-electrode detection unit comprises three-electrode detection arrays, each three-electrode detection array comprises a first reference electrode 21, a first auxiliary electrode 22, a second auxiliary electrode 23, a first working electrode 24 and a second working electrode 25, wherein each electrode comprises a connecting member and a detection member, the detection members are connected to the electrochemical workstation through the connecting members, the two detection members of one reference electrode are respectively located in two detection channels, and the detection member of one auxiliary electrode and the detection member of one working electrode are further arranged in each detection channel. The upper cover plate of the microfluidic chip is coated with a conductor material, a signal is transmitted to the upper cover plate through a connecting piece and then is connected with an electrochemical workstation, wherein a reference electrode is used for providing a stable electrode potential in the detection process, an auxiliary electrode is used for forming a loop with a working electrode in the whole detection process, the working electrode is an area where electrochemical reaction occurs in the detection process, and the electrode potential and potential change can be accurately measured through the three-electrode detection array. Three-electrode detection arrays and two detection channels are arranged, so that six groups of data are actually obtained, and the concentration of a sample to be detected can be more accurately obtained.
As shown in fig. 5, the connection part 26 of the first reference electrode is located between the two detection channels, the connection part 27 of the first auxiliary electrode and the connection part 28 of the first working electrode are located on one side of the two detection channels, and the connection part 29 of the second auxiliary electrode and the connection part 30 of the second working electrode are located on the other side of the two detection channels, so that the arrangement is more reasonable, and the wire arrangement between the rear part and the electrochemical workstation is facilitated.
The length of the detection part 31 of the reference electrode is greater than the sum of the length of the detection part 32 of the auxiliary electrode and the length of the detection part 33 of the working electrode, the transverse area of the detection part of the auxiliary electrode and the transverse area of the detection part of the working electrode are positioned in the transverse area of the detection part of the reference electrode, and the transverse area of the detection part of the auxiliary electrode and the transverse area of the detection part of the working electrode are not overlapped, so that each three-electrode detection array can accurately measure a specific area, and the calculation is more accurate.
Further, as shown in fig. 6, the magnetic field driving module includes eight electromagnetic coils 34 capable of driving the movement of the immunomagnetic beads in the microfluidic chip in multiple directions, each electromagnetic coil is connected to one relay, so that the relay module includes eight relays, and the upper computer controls the on/off of the relay module.
The method for determining the concentration comprises the following steps:
preparing n standard solutions with equal concentration gradients, wherein the concentration of each standard solution is known, passing each standard solution through a detection area, controlling the potential of a three-electrode detection unit to perform one or more repeated scans in a triangular waveform at different time rates, and alternately performing different reduction reactions and oxidation reactions on the electrodes within the potential range to establish n current-potential curves of the standard solutions with different concentrations;
obtaining the signal intensity of each standard solution according to a current-potential curve by utilizing the proportional relation between the corresponding signal intensity of the standard solution and the concentration thereof, wherein the signal intensity can be the maximum current generally, namely, each standard solution corresponds to one maximum current, and establishing corresponding standard curves of different standard solution concentrations and signal intensity sizes through data fitting;
when a sample to be detected with unknown concentration is detected, the concentration of the sample to be detected can be obtained in the standard curve through the signal intensity obtained by the detection of the three-electrode detection unit.
The expression of the concentration of the sample to be detected is as follows:
yt＝a+bxt；
wherein, ytRepresenting the signal intensity, x, of the sample to be testedtWhich is indicative of the concentration of the sample to be tested,
in the formula (I), the compound is shown in the specification,
wherein x isiDenotes the concentration of the i-th standard solution, yiIndicates the signal intensity of the i-th standard solution.
What has been described above is only a preferred embodiment of the present application, and the present invention is not limited to the above embodiment. It is to be understood that other modifications and variations directly derivable or suggested by those skilled in the art without departing from the spirit and concept of the present invention are to be considered as included within the scope of the present invention.
Claims (8)
1. The micro-fluidic chip detection system for food safety rapid detection is characterized by comprising an upper computer, an electrochemical workstation, a relay module, a magnetic field driving module and a micro-fluidic chip, wherein the upper computer is connected to the relay module and the electrochemical workstation, the relay module is connected to the magnetic field driving module, the micro-fluidic chip is installed in the magnetic field driving module, and the micro-fluidic chip is connected to the electrochemical workstation;
the micro-fluidic chip comprises an upper cover plate and a lower cover plate, wherein the upper cover plate and the lower cover plate are bonded together, a micro-fluidic channel is formed in the upper cover plate and comprises a separation area, a washing area, a detection area and a waste liquid area, the separation area is communicated with the detection area through the washing area, the waste liquid area is communicated with the separation area and the detection area, the detection area comprises two detection channels and a three-electrode detection unit, the two detection channels are communicated with the washing area, one end of the three-electrode detection unit is connected to the two detection channels, the other end of the three-electrode detection unit is connected to the electrochemical workstation, and the electrochemical workstation determines the concentration of a sample to be detected in the micro-fluidic.
2. The detecting system according to claim 1, wherein the separating region includes a micro reaction tank and a plurality of injection ports, the injection port for buffer solution, the injection port for immunomagnetic beads and the injection port for sample to be detected are connected to the micro reaction tank, the waste liquid region includes a first waste liquid tank and a second waste liquid tank, a first valve is disposed on a connecting channel between the first waste liquid tank and the micro reaction tank, the washing region includes a washing tank and a washing liquid injection port, a second valve is disposed on a connecting channel between the micro reaction tank and the washing tank, a third valve is disposed on a connecting channel between the washing tank and the detecting region, and the washing region is connected to the second waste liquid tank through the two detecting channels.
3. A test system according to claim 1, wherein the three-electrode test unit comprises three-electrode test arrays, each three-electrode test array comprising a reference electrode, two auxiliary electrodes and two working electrodes, each electrode comprising a connecting member and a test element, the test element being connected to the electrochemical workstation via the connecting member, the two test elements of the one reference electrode being located in the two test channels, respectively, and the test element of one auxiliary electrode and the test element of one working electrode being located in each test channel.
4. A test system according to claim 3, wherein the connection to the reference electrode is located between the two test channels, the connection to the first auxiliary electrode and the connection to the first working electrode are located on one side of the two test channels, and the connection to the second auxiliary electrode and the connection to the second working electrode are located on the other side of the two test channels.
5. A test system according to claim 3 wherein the length of the sensing element of the reference electrode is greater than the sum of the length of the sensing element of the auxiliary electrode and the length of the sensing element of the working electrode, the lateral extent of the sensing element of the auxiliary electrode and the lateral extent of the sensing element of the working electrode are located within the lateral extent of the sensing element of the reference electrode, and the lateral extent of the sensing element of the auxiliary electrode and the lateral extent of the sensing element of the working electrode do not coincide.
6. A testing system according to claim 1, wherein said magnetic field driving module comprises eight electromagnetic coils, said relay module comprises eight relays, each electromagnetic coil is connected to one relay, and said host computer controls the on/off of each relay.
7. The detection system according to claim 1, wherein the determining the concentration of the sample to be detected in the microfluidic chip comprises:
preparing n standard solutions with equal concentration gradients, wherein the concentration of each standard solution is known, enabling each standard solution to pass through a detection area, controlling the potential of a three-electrode detection unit to scan in a triangular waveform at different time rates, and establishing n current-potential curves of the standard solutions with different concentrations;
obtaining the signal intensity of each standard solution according to the current-potential curve, and establishing corresponding standard curves of different standard solution concentrations and signal intensity sizes through data fitting;
when a sample to be detected with unknown concentration is detected, the concentration of the sample to be detected can be obtained in the standard curve through the signal intensity obtained by the detection of the three-electrode detection unit.
8. The detection system according to claim 7, wherein the concentration of the sample to be detected is expressed as:
yt＝a+bxt；
wherein, ytRepresenting the signal intensity, x, of the sample to be testedtWhich is indicative of the concentration of the sample to be tested,
in the formula (I), the compound is shown in the specification,
wherein x isiDenotes the concentration of the i-th standard solution, yiIndicates the signal intensity of the i-th standard solution.
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| CN117625759A (en) * | 2023-12-08 | 2024-03-01 | 精智未来(广州)智能科技有限公司 | Molecular detection method and device based on silicon-based microfluidic chip |
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