CN116445283A - Microfluidic cell culture detection device and preparation method and application thereof - Google Patents
Microfluidic cell culture detection device and preparation method and application thereof Download PDFInfo
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- CN116445283A CN116445283A CN202310420814.XA CN202310420814A CN116445283A CN 116445283 A CN116445283 A CN 116445283A CN 202310420814 A CN202310420814 A CN 202310420814A CN 116445283 A CN116445283 A CN 116445283A
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- 238000004113 cell culture Methods 0.000 title claims abstract description 155
- 238000001514 detection method Methods 0.000 title claims abstract description 108
- 238000002360 preparation method Methods 0.000 title description 5
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 239000002699 waste material Substances 0.000 claims abstract description 12
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 9
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 8
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 238000007877 drug screening Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims abstract description 4
- 238000011160 research Methods 0.000 claims abstract description 4
- 230000005540 biological transmission Effects 0.000 claims description 19
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical group C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims description 12
- 238000009792 diffusion process Methods 0.000 claims description 9
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims description 8
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims description 8
- 238000000835 electrochemical detection Methods 0.000 claims description 7
- 238000012546 transfer Methods 0.000 claims description 7
- 238000002174 soft lithography Methods 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 238000005530 etching Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 73
- 229940079593 drug Drugs 0.000 abstract description 2
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 238000005259 measurement Methods 0.000 abstract 4
- 238000000840 electrochemical analysis Methods 0.000 abstract 1
- -1 polydimethylsiloxane Polymers 0.000 abstract 1
- 238000010586 diagram Methods 0.000 description 4
- 238000001259 photo etching Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 238000004544 sputter deposition Methods 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 238000004115 adherent culture Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000003466 welding Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- 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
- B01L3/502707—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 characterised by the manufacture of the container or its components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to a microfluidic cell culture detection device which is characterized by comprising a microfluidic cell culture detection chip, a supply device, a detection device and a waste liquid collection device; the supply device is communicated with the microfluidic cell culture detection chip and supplies culture medium or liquid medicine to the audience microfluidic cell culture detection chip; the detection device is electrically connected with the microfluidic cell culture detection chip. The method for preparing the microfluid system by using the polydimethylsiloxane for cell culture and electrical measurement can be used for high-throughput cell culture and detection, and can also be used for performing operations such as cell nutrition supply or drug application, and the open cell culture unit can ensure various operations required by conventional cell culture; and the measurement of the multi-unit electric signals can be completed by only needing fewer external electrodes, so that a measurement circuit is simplified, and the measurement efficiency is improved. The invention is very suitable for research work of high-flux drug screening based on cell surface shape, cell impedance and cell electrochemical analysis.
Description
Technical Field
The invention relates to the field of cell culture and detection, in particular to a microfluidic cell culture detection device and a preparation method and application thereof.
Background
The micro-fluidic chip (Microfluidic chip) is a brand new analysis technology developed in the nineties of the last century, has rapid development in recent years, is a development front of the current analysis science field, and has the most basic characteristics and advantages that a plurality of technologies are flexibly combined on a micro platform, so that automation and miniaturization are realized to a great extent, and the consumption of samples is greatly reduced.
Much of the earlier work reported methods of culturing cells in microfluidic systems, but cell culture in microfluidics is typically a closed system, which is inconvenient to apply drugs or to perform various post treatments on cells, compared to conventional open system cultured cells, and few reports of adding open cell culture structures to microfluidic systems; in addition, the cell impedance/electrochemical data are very important detection data, and if the cell impedance/electrochemical data can be coupled with a microfluidic cell culture array, the cell impedance/electrochemical data can be widely applied to the biological research fields such as drug screening and the like, and the cell impedance/electrochemical data are favorable for realizing a screening test with larger flux.
Disclosure of Invention
In order to solve the technical problems, the invention provides a microfluidic cell culture detection device, which comprises a microfluidic cell culture detection chip, a supply device, a detection device and a waste liquid collection device;
the supply device is communicated with the microfluidic cell culture detection chip and supplies culture medium or liquid medicine to the audience microfluidic cell culture detection chip;
the detection device is electrically connected with the microfluidic cell culture detection chip.
In a specific embodiment, the microfluidic cell culture detection chip comprises a microfluidic cell culture supply part and a detection part positioned at the bottom of the microfluidic cell culture supply part;
the downstream path of the microfluidic cell culture supply part comprises a sample injection channel, a concentration generator, a plurality of transmission channels, a cell culture array and a waste liquid outflow channel;
one end of the sample injection channel is communicated with the supply device through a sample injection pipe, the other end of the sample injection channel is communicated with the concentration generator, the concentration generator is communicated with one end of the transmission channel, the other end of the transmission channel is communicated with the waste liquid collecting device, the cell culture array comprises a plurality of cell culture chambers, and each transmission channel corresponds to one row of cell culture chambers of the cell culture array and is communicated with the cell culture chambers.
In a specific embodiment, the transfer channel is annular in shape on the flow path, the cell culture chamber is arranged in the center of the annular shape, the transfer channel is communicated with the cell culture chamber through a diffusion channel, and the sectional area of the diffusion channel is smaller than 1/400 of the sectional area of the transfer channel.
In a specific embodiment, the diffusion channels are arranged radiation symmetrically around the cell culture chamber.
In one embodiment, the detection part comprises one or more layers of detection circuits, and the detection circuits are electrically connected with the detection device through external electrodes.
In a specific embodiment, the detection circuit comprises a cell impedance detection circuit and/or a cell electrochemical detection circuit.
In a specific embodiment, the cell impedance detection circuit includes a cell impedance row circuit and a cell impedance column circuit;
each cell impedance line circuit passes through each cell culture chamber of the corresponding line of the cell culture array, extends out of the cell impedance line electrode to be in contact with the corresponding cell culture chamber, and is electrically connected with one electrode of the impedance signal detection device through one external line electrode;
each cell impedance column circuit passes through each cell culture chamber of a respective column of the cell culture array and extends beyond the cell impedance column electrode to contact the respective cell culture chamber, and each cell impedance row circuit is electrically connected to the other pole of the impedance signal detection device by an external column electrode.
In a specific embodiment, the cell electrochemical detection circuit comprises a cell electrochemical row circuit, a cell electrochemical column circuit, and a reference circuit;
each cell electrochemical row circuit passes through each cell culture chamber of the corresponding row of the cell culture array, extends out of the cell electrochemical row electrode to be in contact with the corresponding cell culture chamber, and is electrically connected with one pole of the cell electrochemical signal detection device through one external row electrode;
each cell electrochemical row circuit passes through each cell culture chamber of a corresponding row of the cell culture array and extends beyond the cell electrochemical row circuit column electrode to contact the corresponding cell culture chamber, and each cell electrochemical row circuit is electrically connected to the other pole of the cell electrochemical signal detection device by an external column electrode;
the reference circuit passes through each cell culture chamber of the cell culture array and extends out of the reference circuit electrode to contact the corresponding cell culture chamber, and the reference circuit is electrically connected to one pole of the cell electrochemical signal detection device through one external electrode.
The microfluidic cell culture detection device comprises a plurality of cell culture chambers, and can culture and detect cells with high flux. The culture medium and the medicine can be supplied into the cell culture chamber through the diffusion channel and the transmission channel, so that nutrition supply and medicine release to cells in the cell culture chamber are realized, and the cells are in a state of zero flow rate. By designing the detection circuit, the impedance signals of the N multiplied by M cell culture units can be measured only by using the N+M electrodes, and the cell electrochemical signals of the N multiplied by M cell culture units can be measured only by using the N+M+1 electrodes. In addition, the cell culture chamber is an open system, and ordinary cell culture operations can be performed through the upper opening. The concentration gradient generator dilutes the medicine input by the sample inlet into a plurality of concentration gradients and transmits the concentration gradients to each cell culture unit through a transmission channel.
The invention also provides a preparation method of the microfluidic cell culture detection chip in the microfluidic cell culture detection device, which is characterized by comprising the following steps:
s1: preparing a PDMS structure through soft lithography and curing;
s2: bonding the PDMS structure with a PDMS substrate to obtain a microfluidic cell culture supply;
s2: etching the electrode layer to obtain a detection part;
s3: and attaching the microfluidic cell culture supply part to the detection part to obtain the microfluidic cell culture detection chip.
The invention also provides application of the microfluidic cell culture detection device in cell biology research or drug screening.
Drawings
FIG. 1 is a schematic structural diagram of a microfluidic cell detection device.
FIG. 2 is a schematic structural diagram of a microfluidic cell culture detection chip.
FIG. 3 is a schematic diagram of a cell impedance detection circuit.
FIG. 4 is a schematic diagram of a cell electrochemical signal detection circuit.
FIG. 5 is an optical micrograph of cells cultured for 15 hours using the apparatus of the present invention.
FIG. 6 shows the results of cell impedance signals detected using the device of the present invention.
The reference numerals in the drawings denote the following components: 1. the microfluidic cell culture detection chip comprises 11 parts of a sample injection channel, 12 parts of a concentration generator, 13 parts of a transmission channel, 14 parts of a cell culture array, 141 parts of a cell culture chamber, 15 parts of a waste liquid outflow channel, 2 parts of a supply device, 21 parts of a sample injection tube, 3 parts of a detection device, 4 parts of a waste liquid collection device.
Detailed Description
The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
1. Structure of microfluidic cell culture detection device
As shown in FIG. 1, the microfluidic cell culture detection device of the present invention comprises a microfluidic cell culture detection chip 1, a supply device 2, a detection device 3, and a waste liquid collection device 4. The supply device 2 supplies a culture medium or a chemical solution to the microfluidic cell culture detection chip 1 through the sample tube and the microfluidic cell culture detection chip 1. The detection device 3 is electrically connected with the microfluidic cell culture detection chip 1.
As shown in fig. 2, the downstream path of the microfluidic cell culture detection chip 1 includes a sample introduction channel 11, a concentration generator 12, a plurality of transfer channels 13, a cell culture array 14, and a waste liquid outflow channel 15. The sample injection channel 11 is communicated with the sample injection tube 21, the other end of the sample injection channel 11 is communicated with the concentration generator 12, the concentration generator 12 is communicated with one end of the transmission channel 13, the other end of the transmission channel 13 is communicated with the waste liquid collecting device 4, the cell culture array 14 comprises a plurality of rows of cell culture chambers 141, each transmission channel 13 corresponds to one row of cell culture chambers 141 of the cell culture array 14, each cell culture chamber 141 is wrapped by the transmission channel 13, and 6 diffusion channels 131 are separated and communicated with the wrapped cell culture chambers 141.
The bottom of the microfluidic cell culture detection chip 1 is etched with a detection circuit 15, including a cell impedance detection circuit and a cell electrochemical detection circuit, which are separated by an insulating layer.
As shown in fig. 3, the cell impedance detection circuit includes cell impedance row circuits and cell impedance column circuits, each cell impedance row circuit passing through each cell culture chamber 141 of a corresponding row of the cell culture array 14 and extending out of the cell impedance circuit row electrode to contact the corresponding cell culture chamber 141, and each cell impedance row circuit may be electrically connected to one pole of the detection device through an external row electrode. Each cell impedance column circuit passes through each cell culture chamber 141 of a corresponding column of cell culture array 14 and extends beyond the cell impedance column electrodes to contact the corresponding cell culture chamber 141, and each cell impedance row circuit is electrically connectable to the other pole of the detection device via an external column electrode.
As shown in fig. 4, the cell electrochemical detection circuit comprises a cell electrochemical row circuit, a cell electrochemical column circuit and a reference circuit; each cell electrochemical row circuit passes through each cell culture chamber of the corresponding row of the cell culture array, extends out of the cell electrochemical row electrode to be in contact with the corresponding cell culture chamber, and is electrically connected with one pole of the cell electrochemical signal detection device through one external row electrode; each cell electrochemical row circuit passes through each cell culture chamber of a corresponding row of the cell culture array and extends beyond the cell electrochemical row circuit column electrode to contact the corresponding cell culture chamber, and each cell electrochemical row circuit is electrically connected to the other pole of the cell electrochemical signal detection device by an external column electrode; the reference circuit passes through each cell culture chamber of the cell culture array and extends out of the reference circuit electrode to contact the corresponding cell culture chamber, and the reference circuit is electrically connected to one pole of the cell electrochemical signal detection device through one external electrode.
2. Preparation of microfluidic cell culture detection chip
1) The template is shown in fig. 2 by drawing with L-Edit, wherein the width of the sample injection channel 11 is 300 micrometers, the width of the 4 concentration gradient generators 12 and the 4 transmission channels 13 is 200 micrometers, each transmission channel 13 surrounds four circular rings on the path, an open circular hole, namely a cell culture chamber 141, is arranged in the center of each circular ring, and the circular rings of the transmission channels 13 are communicated with the cell culture chamber 141 through six diffusion channels 131 with the height of 5 micrometers.
2) A 50 micron high mold was made on a clean silicon wafer by soft lithography using liquid PDMS (a: b=10: 1) The PDMS was poured onto a mold to a height of about 5mm and cured in an 80 degree oven for 30 minutes. And (3) after cutting, drilling holes at the corresponding inlet and outlet, bonding the PDMS with a clean slide coated with the PDMS in a spin mode, removing the whole PDMS, and perforating in the cell culture chamber area to form an open round hole for later use.
3) Cell impedance detection circuit templates were plotted using L-Edit plot as shown in fig. 3, and cell electrochemical detection circuit templates were shown in fig. 4. The impedance measuring electrode width was 365 microns and the electrode spacing was 310 microns. The electrochemical measurement electrode width was 365 microns, the distance between the common reference electrode and the opposing electrode at the same layer was 310 microns, and the distance between the electrodes at different layers was 140 microns.
4) Preparing a first layer electrode on a clean glass sheet by a photoetching, sputtering and lift-off method, preparing an insulating layer by photoetching, preparing a second layer electrode by repeating photoetching, sputtering and lift-off, finally welding a lead and an interface, and aligning and attaching the PDMS microfluidic channel prepared in the step 2) to an electrode substrate to obtain the microfluidic cell culture detection chip.
3. Use example of microfluidic cell culture detection device
HeLa cells were cultured in DMEM medium using pre-shaking to be uniform and diluted to a suitable concentration, which was approximately 10-4 cells/ml.
Placing the chip in a vacuum environment for about 10 minutes, and injecting a culture medium into the chip through a sample injection hole to fill the chip with a concentration gradient generator, a transmission channel and a diffusion channel; cells were dropped into the cell culture chamber from above, and the cells were subjected to adherent culture. After 15 hours of adherence, microscopic examination of the microfluidic cell culture detection chip under an optical microscope was carried out, as shown in FIG. 5, and it was found that the adherence of cells was good.
The cell impedance signal detection results obtained by connecting the external row electrode and the external column electrode of the cultured microfluidic cell culture detection chip with the cell impedance detection device are shown in fig. 6. Therefore, the microfluidic cell culture detection device can effectively detect cell impedance signals and electrochemical signals in a cell culture chamber.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (10)
1. The microfluidic cell culture detection device is characterized by comprising a microfluidic cell culture detection chip, a supply device, a detection device and a waste liquid collection device;
the supply device is communicated with the microfluidic cell culture detection chip and supplies culture medium or liquid medicine to the audience microfluidic cell culture detection chip;
the detection device is electrically connected with the microfluidic cell culture detection chip.
2. The microfluidic cell culture detection device according to claim 1, wherein the microfluidic cell culture detection chip comprises a microfluidic cell culture supply part and a detection part positioned at the bottom of the microfluidic cell culture supply part;
the downstream path of the microfluidic cell culture supply part comprises a sample injection channel, a concentration generator, a plurality of transmission channels, a cell culture array and a waste liquid outflow channel;
one end of the sample injection channel is communicated with the supply device through a sample injection pipe, the other end of the sample injection channel is communicated with the concentration generator, the concentration generator is communicated with one end of the transmission channel, the other end of the transmission channel is communicated with the waste liquid collecting device, the cell culture array comprises a plurality of cell culture chambers, and each transmission channel corresponds to one row of cell culture chambers of the cell culture array and is communicated with the cell culture chambers.
3. The microfluidic cell culture detection device according to claim 2, wherein the transfer channel is formed in a ring shape on a flow path thereof, the cell culture chamber is provided in a center of the ring shape, the transfer channel communicates with the cell culture chamber through a diffusion channel having a cross-sectional area smaller than 1/400 of a cross-sectional area of the transfer channel.
4. The microfluidic cell culture detection device of claim 3, wherein the diffusion channels are radially symmetrically disposed about the cell culture chamber.
5. The microfluidic cell culture detection device according to any one of claims 2 to 4, wherein the detection section comprises one or more layers of detection circuits electrically connected to the detection device via external electrodes.
6. The microfluidic cell culture detection device of claim 5, wherein the detection circuit comprises a cell impedance detection circuit and/or a cell electrochemical detection circuit.
7. The microfluidic cell culture detection device of claim 6, wherein the cell impedance detection circuit comprises a cell impedance row circuit and a cell impedance column circuit;
each cell impedance line circuit passes through each cell culture chamber of the corresponding line of the cell culture array, extends out of the cell impedance line electrode to be in contact with the corresponding cell culture chamber, and is electrically connected with one electrode of the impedance signal detection device through one external line electrode;
each cell impedance column circuit passes through each cell culture chamber of a respective column of the cell culture array and extends beyond the cell impedance column electrode to contact the respective cell culture chamber, and each cell impedance row circuit is electrically connected to the other pole of the impedance signal detection device by an external column electrode.
8. The microfluidic cell culture detection device of claim 6, wherein the cell electrochemical detection circuit comprises a cell electrochemical row circuit, a cell electrochemical column circuit, and a reference circuit;
each cell electrochemical row circuit passes through each cell culture chamber of the corresponding row of the cell culture array, extends out of the cell electrochemical row electrode to be in contact with the corresponding cell culture chamber, and is electrically connected with one pole of the cell electrochemical signal detection device through one external row electrode;
each cell electrochemical row circuit passes through each cell culture chamber of a corresponding row of the cell culture array and extends beyond the cell electrochemical row circuit column electrode to contact the corresponding cell culture chamber, and each cell electrochemical row circuit is electrically connected to the other pole of the cell electrochemical signal detection device by an external column electrode;
the reference circuit passes through each cell culture chamber of the cell culture array and extends out of the reference circuit electrode to contact the corresponding cell culture chamber, and the reference circuit is electrically connected to one pole of the cell electrochemical signal detection device through one external electrode.
9. The method for manufacturing a microfluidic cell culture detection chip in a microfluidic cell culture detection device according to claims 1 to 8, comprising the steps of:
s1: preparing a PDMS structure through soft lithography and curing;
s2: bonding the PDMS structure with a PDMS substrate to obtain a microfluidic cell culture supply;
s2: etching the electrode layer to obtain a detection part;
s3: and attaching the microfluidic cell culture supply part to the detection part to obtain the microfluidic cell culture detection chip.
10. Use of the microfluidic cell culture detection device according to claims 1-8 in cell biology research or drug screening.
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| CN202310420814.XA CN116445283A (en) | 2023-04-19 | 2023-04-19 | Microfluidic cell culture detection device and preparation method and application thereof |
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| CN202310420814.XA CN116445283A (en) | 2023-04-19 | 2023-04-19 | Microfluidic cell culture detection device and preparation method and application thereof |
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