CN113502225B - Cell culture pore plate and cell culture on-line detection device provided with same - Google Patents
Cell culture pore plate and cell culture on-line detection device provided with same Download PDFInfo
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- CN113502225B CN113502225B CN202110837008.3A CN202110837008A CN113502225B CN 113502225 B CN113502225 B CN 113502225B CN 202110837008 A CN202110837008 A CN 202110837008A CN 113502225 B CN113502225 B CN 113502225B
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- 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
- C12M23/12—Well or multiwell plates
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- 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
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- 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
- C12M23/48—Holding appliances; Racks; Supports
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- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/06—Plates; Walls; Drawers; Multilayer plates
- C12M25/08—Plates; Walls; Drawers; Multilayer plates electrically charged
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- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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Abstract
The invention relates to the technical field of biochemical detection and discloses a cell culture pore plate integrating an electrical stimulation and measurement technology, which comprises a pore plate base, wherein a plurality of holes are formed in the base, an electrode body is arranged at the bottom of each hole, each electrode body comprises a first electrode and a second electrode, a plurality of poles are formed on opposite sides of each electrode body, the poles are arranged at intervals, each pole consists of a repeated regular structure, patterns between adjacent poles are staggered, the shape of the electrode body after arrangement is matched with the shape of the bottom of each hole, the holes are arranged in an array manner, and the first electrode and the second electrode in each hole are led out to two ends of the pore plate base. The invention also discloses a cell culture on-line detection device matched with the cell culture pore plate. The invention realizes the control and more accurate detection of the cell state and growth condition in the cell culture process.
Description
Technical Field
The invention relates to the technical field of biochemical detection, in particular to a cell culture pore plate for online detection of cell growth and a cell culture online detection device provided with the cell culture pore plate.
Background
Drug development and development of cell therapy technologies, there is an urgent need to have on-line, in-situ, non-destructive and continuous cell detection technologies for quantitative analysis of cell growth status, growth trend and number during cell culture. The technology can be widely used for drug screening, drug toxicity analysis, anti-tumor drug killing efficiency analysis, stem cell culture process optimization, stem cell differentiation state monitoring, carT cell treatment method verification and the like.
Conventional cell viability assays require that the cells be labeled and light intensity measurements be taken after incubation to obtain the desired data, i.e., CI (known as CellIndex). This results in a long experimental flow, and only a few or even one data can be obtained in one experiment, which is inefficient, and the data analysis is required to be performed through multiple experiments, so as to achieve the requirement of the data volume of the graph drawing and the data analysis.
Measurement of the state of cell growth based on the electrical properties of cells is currently one of the most important methods for online detection of cells. Typically, the technique is used to obtain electrical characteristics of the cell by applying an excitation voltage, for example, measuring the resistance, capacitance, or impedance of the cell. Currently, there is a technology for obtaining cell growth information by culturing cells on the surface of an electrode and utilizing the influence of the cells on the impedance of the electrode surface. The method generally adopts linear interdigital electrodes for measurement, and the excitation voltage is sinusoidal with fixed frequency, so that when the method is used for measuring different types of cells, the detection sensitivity is different due to the difference of cell orientation, size and shape, namely, the design of interdigital electrode units in a single electrode body branch is difficult to meet the high-sensitivity detection of all specified cell growth.
Obviously, in order to achieve higher sensitivity for specific cell measurement, it is necessary to improve or redesign parameters such as shape, pitch, material, thickness, etc. of the interdigital electrodes, and develop an instrument system with high accuracy and stability for measuring cell growth on the electrode surface, so as to obtain a dynamic change curve of cell growth state, thereby being widely used in cell growth experiments.
Disclosure of Invention
The invention aims to solve the problems and provide a cell culture pore plate and a cell culture on-line detection device, which realize the control of cell growth and the detection of cell growth state in the cell culture process.
The technical scheme adopted by the invention is as follows:
The utility model provides a cell culture orifice plate, its characterized in that includes the orifice plate base, sets up a plurality of holes on the base, and the hole bottom sets up the electrode body, the electrode body includes first electrode and second electrode, and first electrode and second electrode all form many branches to the offside, and branch interval arrangement forms the interdigital form, and every branch is by repeated arrangement's geometric pattern unit structure, by wire connection constitution, and the pattern between adjacent branches is staggered arrangement, and the electrode body shape after arranging matches with hole bottom shape, a plurality of hole array arrangement, the interface at orifice plate base both ends is drawn forth to the first electrode and the second electrode of every hole.
Further, the number of the poles is 16 to 100; the hole array is 16-384 holes.
Further, the cross section of the hole is one of round, square, rectangular or hexagonal, the bottom material of the hole is one of glass, quartz, polytetrafluoroethylene, polyethylene terephthalate, polymethyl methacrylate and polyvinyl chloride, the electrode body structure is processed at the bottom of the hole through a magnetron sputtering or vapor plating process, the outer layer of the electrode body is a conductive material with the thickness of 10nm to 500nm, and the conductive material of the outer layer of the electrode body is one of gold, platinum, rhodium, ruthenium, silver chloride, polypyrrole or poly 3, 4-ethylenedioxythiophene.
Further, the cells are cultivated on the bottom of the hole in an adherence way, and the individual cells are covered on the electrode body area and span between the first electrode and the second electrode in the cultivation process.
The cell culture on-line detection device is characterized by comprising a base and a cell culture pore plate, wherein a pore plate groove is formed in the base, the cell culture pore plate is arranged in the pore plate groove, a control circuit is arranged in the base, an electrode body on the cell culture pore plate is connected to the control circuit, the control unit comprises a main control circuit, a signal reference source, a multiplexer and a signal measurement circuit, the main control circuit controls the reference signal source to generate excitation signals, the multiplexer is respectively connected to each first electrode on the cell culture pore plate, the multiplexer is controlled by the main control circuit to connect the signal input end of the signal measurement circuit with one of the first electrodes, the signal output end of the signal measurement circuit is connected with all the second electrodes, the signal reference source is connected with the signal measurement circuit, the signal measurement circuit is used for conditioning the output signals of the signal reference source, outputting the excitation signals to all the second electrodes, and the first electrodes selected by the multiplexer form a loop for applying electric stimulation to cells or measuring the electrical properties of the cells, and monitoring the growth state of the cells.
Further, the control unit periodically generates pulses of 10mV to 10V voltage amplitude, 1Hz to 1kHz frequency, and 1% to 50% duty cycle on the second electrode, which pulses adjust the arrangement and state of cells on the electrode surface.
Further, the control unit measures current amplitude and phase change data of the electrode, performs normalization processing to obtain cell growth state indexes, and analyzes the change of the cell growth state indexes along with time to obtain cell growth trend information.
Further, the excitation signal output to the second electrode is a voltage signal having a waveform such as a sine wave, a square wave, a triangular wave, or a pulse, and the amplitude of the excitation signal output to the second electrode is 1mV to 150mV.
Further, the control unit further comprises a calibration circuit connected to the signal measurement circuit through the multiplexer, the control circuit stores calibration data, the signal measurement circuit calibrates the signal measurement result through the signal calibration circuit, and stores the calibration result for compensating the subsequent measurement data.
Further, the control circuit is also connected with a chip in-situ detection circuit, and the chip in-situ detection circuit is connected to the cell culture pore plate, detects the loading state of the cell culture pore plate and feeds back to the control circuit.
Further, the main control circuit is also connected to a storage unit for storing the detection data.
Further, the main control circuit is connected with the upper computer software through the communication circuit, transmits the data acquired by the measuring circuit to the upper computer, and receives an instruction sent by the upper computer to execute operation.
Further, the main control circuit is also connected with a status indicator lamp.
Further, a flip cover is arranged on the pore plate groove, when the flip cover is closed, the cell culture pore plate is fixed in the pore plate groove and the flip cover, and a detachable transparent observation window is arranged above the cell culture pore plate.
The beneficial effects of the invention are as follows:
(1) The electrode design of the cell culture pore plate can be designed in a parameterization way by adjusting the width and the angle of the structural unit aiming at cells with specific size, shape and physicochemical properties so as to improve the detection sensitivity;
(2) The electrodes are staggered in a diamond or hexagonal unit structure, so that the detection sensitivity of cells with different sizes and spatial orientations is improved;
(3) The electrode geometry is beneficial to processing of various processes including photoetching, laser etching, sputtering and the like;
(4) The cell culture on-line detection device selects channels through a multiplexer, and can measure any channel in single detection;
(5) Regulating cell growth by applying an electrical stimulation signal to the cell;
(6) And (5) carrying out automatic calibration compensation on the measurement result through a calibration circuit.
Drawings
FIG. 1 is a schematic perspective view of a cell culture well plate of the present invention;
FIG. 2 is a schematic plan view of a circular electrode;
FIG. 3 is an enlarged view of a bent portion of the branch corresponding to FIG. 2;
FIG. 4 is a schematic plan view of an electrode having a rectangular shape;
FIG. 5 is an enlarged view of a diamond corresponding to the branch of FIG. 4;
FIG. 6 is a schematic plan view of a diamond-shaped electrode;
FIG. 7 is a schematic plan view of a circular shaped electrode;
FIG. 8 is an enlarged hexagonal view of the branch corresponding to FIG. 7;
FIG. 9 is a schematic perspective view of an on-line cell culture detection apparatus;
FIG. 10 is a schematic diagram showing a three-dimensional structure of a cell culture on-line detecting device with a flip cover opened;
FIG. 11 is a block diagram of a control unit of the cell culture on-line detection device;
FIG. 12 is a graph of growth of cells of multiple channels before and after intervention.
Detailed Description
The following describes in detail the embodiments of the cell culture well plate and the cell culture on-line measuring device according to the present invention with reference to the accompanying drawings.
Referring to fig. 1, a cell culture well plate 1 comprises a well plate base 2, a plurality of holes 3 are arranged on the base 2, and the number of the holes 3 is 16 to 384 holes which are arranged in an array, and the holes are generally 16, 48, 96, 384 and other holes.
Referring to fig. 2, fig. 2 is a bottom shape of a form of hole 3, an electrode body 4 is disposed at the bottom of each hole 3, the electrode body 4 includes a first electrode 5 and a second electrode 6, the first electrode 5 and the second electrode 6 each form a plurality of branches 7 toward opposite sides, and the number of branches 7 is preferably 16 to 100, which is determined according to the size of cultured cells. The poles 7 are arranged at intervals, each pole 7 is of a zigzag structure, the shape of the arranged electrode body 4 is matched with the shape of the bottom of the hole 3, and the electrode body 4 is also round according to the round hole 3 in the figure.
Referring to fig. 3, the width of the zigzag bending part of the branch electrode 7 is d, the bending angle omega is determined according to the actual cell type, and specific values of the width d and the angle omega are determined, so that the contact position between the cell growth process and the branch electrode 7 is optimally matched.
Referring to fig. 4, fig. 4 is an electrode shape of the bottom of the rectangular hole 3. The first electrode 5 and the second electrode 6 are enclosed into a rectangular shape, the number of the branch poles 7 is 16, the two branch poles are formed by connecting a plurality of diamonds, and the diamonds on the adjacent branch poles 7 are staggered.
Referring to fig. 5, ω=133°, d=100 μm in the corresponding diamond parameters.
Referring to fig. 6, fig. 6 shows the bottom electrode shape of the diamond-shaped holes 3. The first electrode 5 and the second electrode 6 are in a rhombic shape, 20 branches 7 are arranged from the top to the bottom of the rhombic electrodes, each branch 7 is formed by connecting a plurality of rhombuses, and the rhombuses on the adjacent branches 7 are staggered. Corresponding to the diamond parameters in fig. 5, ω=70°, d=50 μm.
The two electrodes of the electrode body 4 are respectively led out to one end of the orifice plate base 2, the two electrodes of the plurality of holes 3 are respectively arranged at two ends of the orifice plate base 2, and the lead-out wires at the two ends of the orifice plate base 2 are arranged as contacts on the lower surface of the orifice plate base 2.
On this basis, the cross-sectional shape of the holes 3 may be circular, square, rectangular or hexagonal. Referring to fig. 7 and 8, the first electrode 5 and the second electrode 6 are enclosed into a circular shape, the number of the branch electrodes 7 is 16, and each branch electrode is formed by connecting a plurality of regular hexagons, and the regular hexagons on the adjacent branch electrodes 7 are staggered. The spacing between adjacent regular hexagons in all directions is equal. The diameter r of the circumscribed circle of the regular hexagon is set differently according to different cells.
The bottom material of the hole 3 can be glass, polyethylene terephthalate, polymethyl methacrylate, polyvinyl chloride and other materials, the electrode body structure is processed at the bottom of the hole 3 through a magnetron sputtering or vapor deposition process, the outer layer of the electrode body is made of conductive materials with the thickness of 10nm to 500nm, and the conductive materials of the outer layer of the electrode body are gold, platinum, rhodium, ruthenium, silver chloride, polypyrrole or poly 3, 4-ethylenedioxythiophene.
During measurement, cells can be cultivated on the bottom of the hole 3 in an adherence way, and most of the cells are covered on the electrode body area in the cultivation process and spanned between the first electrode 5 and the second electrode 6 by adjusting the angle and the width of the repeated regular structural units of the branch electrode.
Referring to fig. 9 and 10, the cell culture well plate 1 is installed in the cell culture on-line detecting device 8. The cell culture on-line detecting device 8 is characterized in that an orifice plate groove 9 is formed in the base of the cell culture on-line detecting device 8, a cell culture electrode channel orifice plate cell culture orifice plate 1 is arranged in the orifice plate groove 9, electrode contacts are arranged at two ends of the orifice plate groove 9, and after the cell culture orifice plate 1 is installed, contacts at two ends of the orifice plate base 2 are electrically connected with the electrode contacts at two ends of the orifice plate groove 9 in a contact mode and used for electrically connecting electrodes in the cell culture orifice plate 1 with the cell culture on-line detecting device 8.
The cell culture pore plate 1 is sealed in the pore plate groove 9 and the flip cover 10 when the flip cover 10 is closed, and a transparent observation window is arranged above the flip cover 10. After adding cells and relevant culture auxiliary materials into the cell culture pore plate 1, the flip cover 10 is closed, and the whole cell culture on-line detection device 8 is placed into an incubator for culture and measurement.
Referring to fig. 11, a control circuit is arranged in the base, electrode contacts at two ends of the pore plate groove are connected to the control unit, the control unit comprises a main control circuit, a signal reference source, a multiplexer and a signal measuring circuit, the main control circuit controls the reference signal source to generate an excitation signal, the multiplexer is connected to an electrode on the cell culture pore plate, the multiplexer is connected with the signal measuring circuit, and the signal measuring circuit performs signal measurement on an electrode passage selected by the multiplexer.
The control unit further comprises a calibration circuit which is connected to the signal measurement circuit through the multiplexer, the control circuit stores calibration data, and the signal measurement circuit calibrates the signal measurement result through the signal calibration circuit. The control circuit is also connected with a chip in-situ detection circuit which is connected to the cell culture pore plate and used for detecting the loading state of the cell culture pore plate and feeding back the cell culture pore plate to the control circuit.
The main control circuit is also connected to a storage unit for storing the detection data. The main control circuit is connected to the upper computer software through the communication circuit.
The main control circuit controls the reference signal source to generate excitation signals with certain frequency, waveform, amplitude and phase. Comprising the following steps: sinusoidal, pulsed, triangular and square wave. The signal measuring circuit amplifies and outputs the excitation signal, simultaneously monitors the output signal and the current signal passing through the cell culture pore plate, and obtains cell response signals under different frequencies by adopting a frequency scanning mode in single measurement for subsequent analysis.
The channels are selected by the multiplexer, and the contact probes are connected with the sensing electrodes of the cell culture pore plate, so that any channel can be measured in a single detection. The multiplexer can connect the signal measurement circuit with the calibration circuit for automatic calibration of the signal measurement result; the calibration data is stored by the main control circuit and is used for compensating the result of the subsequent actual measurement. The chip in-place detection is used for detecting whether the sensor array chip is loaded correctly in real time.
Status indicators are used to indicate current device status including standby, running, malfunction, calibration.
The main control circuit is communicated with the upper computer through the communication circuit, and the upper computer sends the measurement result to the upper computer software in real time through an interface 11 on the cell culture on-line detection device 8; if the communication of the upper computer software is interrupted in the running process, the measurement data are stored in a storage unit of the equipment and are sent to the upper computer after the communication is recovered.
The measurement process is as follows: the electrode body on the cell culture pore plate is connected to the control circuit, the electric stimulation can be applied to the cells or the electrical property of the cells can be measured through the control circuit, the control unit comprises a main control circuit, a signal reference source, a multiplexer and a signal measuring circuit, the main control circuit controls the reference signal source to generate excitation signals, the multiplexer is respectively connected to each first electrode on the cell culture pore plate, the multiplexer is controlled by the main control circuit to connect the signal input end of the signal measuring circuit with one of the first electrodes, the signal output end of the signal measuring circuit is connected with all the second electrodes, the signal reference source is connected with the signal measuring circuit, the signal measuring circuit regulates the output signals of the signal reference source, and then outputs the excitation signals to all the second electrodes, and the first electrodes selected by the multiplexer form a loop to realize the electric stimulation or the signal measurement of the cells.
The purpose of the electrical stimulation of the cells is to adjust the arrangement and state of the cells on the electrode surface by external action, which is represented by a series of pulses of 10mV to 10V voltage amplitude, 1Hz to 1kHz frequency and 1% to 50% duty cycle continuously generated at a certain period.
The cell signal measurement aims to estimate the number of cells attached to the bottom of an orifice plate by measuring the total electrical quantity of cells covered on the electrodes so as to infer the change trend of cell growth, and the principle of the cell signal measurement is to output a series of excitation signals with different frequencies on a second electrode and simultaneously measure the current amplitude and phase change on a selected first electrode, normalize the measured current amplitude and phase change data by taking the signal in an initial cell culture state as a reference, take the measured current amplitude and phase change data as a cell growth state index, record the cell growth state index at certain time intervals, and draw a curve of 'cell growth state index-time' so as to visualize the cell growth trend.
The control unit controls the excitation signal output to the second electrode to be a voltage signal having a waveform such as a sine wave, a square wave, a triangular wave, a pulse, or the like, and the amplitude of the excitation signal output to the second electrode is 1mV to 150mV.
The visualization of the cell growth trend can be used for optimizing a stem cell culture process, analyzing the killing effect of cancer cells of anticancer drugs, inhibiting the microorganism by antibiotics and monitoring the in-vitro culture condition of functional cells.
In application, the cell culture on-line detection device of the invention is added with cells to be detected and culture solution, then the cell culture on-line detection device is put into an incubator, the growth, adaptability and behavior of the cells are studied in real time, the invasion and migration of the cells are monitored, the cytotoxicity reaction caused by a wide molecular target is monitored, the differentiation process of stem cells into somatic cells is captured in real time, and the adhesion and expansion of the cells are continuously monitored. Can be used for cancer immunotherapy.
Referring to FIG. 12, FIG. 12 is a SKOV3 ovarian cancer tumor cell killing experiment. 20000 tumor cells were spread on the bottom of the cell culture well plate one day in advance, and the target ratio was 2.5 in the corresponding well plate the next day: 1 and 5:1, 4 total groups of NC-T and CAR-T cells, wherein 2.5:1 is 50000 cells, 5:1 is 100000 cells. Curve 1 and curve 2 are effective target ratios of 2.5, respectively: 1 and 5:1 in a blank NC-T experimental group; curve 3 and curve 4 are effective target ratios of 2.5, respectively: 1 and 5: 1. Experimental results indicate that CAR-T cells can significantly kill tumor cells.
As can be seen from the figure, cells grew normally before killing cancer cells, CAR-T cells were added to the channels of the CAR-T cell experimental group at 1200 minutes, and then the cell growth trend (Cellindex) of cancer cells in the channels was decreased, while cells of the blank group were not affected.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. An on-line cell culture detection device, which is characterized in that: comprises a base and a cell culture pore plate, wherein a pore plate groove is arranged on the base,
The cell culture pore plate comprises a pore plate base, a plurality of holes are arranged on the pore plate base, an electrode body is arranged at the bottom of each hole, the electrode body comprises a first electrode and a second electrode, a plurality of branches are formed on opposite sides of the first electrode and the second electrode, the branches are arranged between each other to form an interdigital form, each branch is formed by a geometrical pattern unit structure which is repeatedly arranged and is formed by connecting wires, patterns between adjacent branches are arranged in a staggered way, the shape of the arranged electrode body is matched with the shape of the bottom of each hole, the holes are arranged in an array way, the first electrode and the second electrode of each hole are led out to interfaces at two ends of the pore plate base, cells are cultivated at the bottom of each hole in an adherence way, cells are covered in the electrode body area in the cultivation process and span between the first electrode and the second electrode,
The cell culture pore plate is arranged in the pore plate groove, a control unit is arranged in the base, an electrode body on the cell culture pore plate is connected to the control unit, the control unit comprises a main control circuit, a signal reference source, a multiplexer and a signal measuring circuit, the main control circuit controls the reference signal source to generate excitation signals, the multiplexer is respectively connected to each first electrode on the cell culture pore plate, the multiplexer connects the signal input end of the signal measuring circuit with one of the first electrodes under the control of the main control circuit, the signal output end of the signal measuring circuit is connected with all the second electrodes, the signal reference source is connected with the signal measuring circuit, and after the signal measuring circuit conditions the signal output signal of the signal reference source, the control unit also comprises a calibration circuit, the calibration circuit is connected to the signal measurement circuit through the multiplexer, the control unit stores calibration data, the signal measurement circuit calibrates the signal measurement result through the signal calibration circuit and stores the calibration result for compensating the subsequent measurement data, the main control circuit is also connected with a chip in-place detection circuit, the chip in-place detection circuit is connected to the cell culture pore plate, and the cell culture pore plate loading state is detected and fed back to the control unit.
2. The cell culture on-line detection apparatus of claim 1, wherein: the number of the branches is 16 to 100; the hole array is 16-384 holes.
3. The cell culture on-line detection apparatus of claim 1, wherein: the cross section of the hole is one of round, square, rectangular or hexagonal, the bottom material of the hole is one of glass, quartz, polytetrafluoroethylene, polyethylene terephthalate, polymethyl methacrylate and polyvinyl chloride, the structure of the electrode body is processed at the bottom of the hole through a magnetron sputtering or vapor plating process, the outer layer of the electrode body is a conductive material with the thickness of 10nm to 500nm, and the conductive material of the outer layer of the electrode body is one of gold, platinum, rhodium, ruthenium, silver chloride, polypyrrole or poly 3, 4-ethylenedioxythiophene.
4. The cell culture on-line detection apparatus of claim 1, wherein: the control unit periodically generates pulses of 10mV to 10V voltage amplitude, 1Hz to 1kHz frequency and 1% to 50% duty cycle on the second electrode, which pulses adjust the arrangement and state of cells on the electrode surface.
5. The cell culture on-line detection apparatus of claim 1, wherein: the control unit measures current amplitude and phase change data on the electrode, performs normalization processing, serves as a cell growth state index, analyzes the change of the cell growth state index along with time, and obtains cell growth trend information.
6. The cell culture on-line detection apparatus of claim 1, wherein: the excitation signal output to the second electrode is a voltage signal with waveforms such as sine wave, square wave, triangular wave and pulse, and the amplitude of the excitation signal output to the second electrode is 1mV to 150mV.
7. The cell culture on-line detection apparatus according to any one of claims 1 to 6, wherein: the main control circuit is also connected to a storage unit, and the storage unit is used for storing detection data.
8. The cell culture on-line detection apparatus according to any one of claims 1 to 6, wherein: the main control circuit is connected with the upper computer software through the communication circuit, transmits the data acquired by the measuring circuit to the upper computer, and receives an instruction sent by the upper computer to execute operation.
9. The cell culture on-line detection apparatus according to any one of claims 1 to 6, wherein: and the main control circuit is also connected with a status indicator lamp.
10. The cell culture on-line detection apparatus according to any one of claims 1 to 6, wherein: the cell culture pore plate is characterized in that a flip cover is arranged on the pore plate groove, when the flip cover is closed, the cell culture pore plate is fixed in the pore plate groove and the flip cover, and a detachable transparent observation window is arranged above the cell culture pore plate.
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