CN111289819B - Integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation - Google Patents

Abstract

The invention discloses an integrated recording and regulating system for measuring intracellular electrical signals by electroporation of myocardial cells, comprising an upper computer, a PCB substrate, a hollow platinum nanotube array sensor fixed on the PCB substrate, a sensor electrical signal conditioning circuit, and an electroporation Signal output line and signal acquisition card. The hollow platinum nanotube array sensor includes a PCB board, a working electrode and a reference electrode. The working electrode is connected to the input end of the sensor electrical signal conditioning circuit; the reference electrode is grounded. A cylinder is fixed on the working electrode and the reference electrode as a cell culture chamber. The microelectrode consists of a PET film and an array of hollow platinum nanotubes with a diameter of 0.4-1 μm and a length of 0.5-2 μm grown on the PET film. The system of the invention adopts the minimally invasive electroporation method to record the intracellular electrical signals of the cardiomyocytes while ensuring that the basic activity of the cardiomyocytes is not affected. Long record.

Description

An integrated recording and regulation system for measuring intracellular electrical signals by electroporation of cardiomyocytes

technical field

The invention relates to an automatic recording device for intracellular electrical signals of cardiomyocytes, in particular to an integrated recording and regulating system capable of performing myocardial electroporation and intracellular electrical signals.

Background technique

Cardiomyocytes are the basic units of the heart, which can rhythmically generate action potentials and then produce mechanical contractions. These are the basic biological functions of the heart for pumping blood. Therefore, cardiomyocyte electrophysiology research plays a vital role in the field of cardiology. effect. At present, the patch clamp technique is a commonly used method for recording intracellular action potential signals of cardiomyocytes. Although this technique can effectively record standard action potential signals, its invasive detection principle has great damage to cardiomyocytes. , cannot carry out long-term intracellular electrophysiological studies of cardiomyocytes. The extracellular electrical signal recording of cardiomyocytes based on microelectrode arrays can non-invasively record extracellular electrical signals for a long time, and these signals can reflect the electrophysiological state of cardiomyocytes to a certain extent. However, the extracellular electrical signals recorded based on microelectrode arrays It is a deformed action potential, which becomes a hindrance to the in-depth study of intracellular electrophysiology of cardiomyocytes.

SUMMARY OF THE INVENTION

The purpose of the present invention is to solve the problem that the existing technology cannot stably record the intracellular electrical signals of cardiomyocytes for a long time, and develops an integrated recording and regulation system of myocardial electroporation and intracellular electrical signals based on a conductive nano-needle tube array. The perforation method and the label-free signal recording method automate the long-term recording of the intracellular electrical signals of cardiomyocytes.

The purpose of this invention is to realize through the following technical solutions:

An integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes, comprising an upper computer, a PCB substrate, a hollow platinum nanotube array sensor fixed on the PCB substrate, a sensor electrical signal conditioning circuit, an electroporation signal output line and Signal acquisition card.

The hollow platinum nanotube array sensor includes a PCB board, a working electrode and a reference electrode. The electrode end of the working electrode is 6.5 mm long and 20-100 μm wide, and the connecting end is 5 mm long and 2 mm wide; the electrode end of the reference electrode is 4.5 mm long. mm, the width is 20-100μm, the length of the connecting end is 8mm, and the width is 2mm. The working electrode is connected with the input end of the sensor electrical signal conditioning circuit; the reference electrode is grounded. The working electrode and the reference electrode are fixed on the PCB board through the connecting end. A cylinder is fixed on the working electrode and the reference electrode as a cell culture chamber.

The working electrode and the reference electrode are both composed of a PET film and a hollow platinum nanotube array with a diameter of 0.4-1 μm and a length of 0.5-2 μm grown on the PET film.

The upper computer is used to control the output of electroporation signals, and to display and record the data obtained by the hollow platinum nanotube array sensor test.

The sensor electrical signal conditioning circuit is composed of a first-stage amplifying circuit, a low-pass filter, an RC blocking circuit and a second-stage amplifying circuit connected in sequence, and the first-stage amplifying circuit and the second-stage amplifying circuit are both proportional amplifiers of the same phase. ;The RC blocking circuit is composed of a 10μF capacitor C7 and a resistor R16 and a resistor R17 with a resistance value of 30kΩ. The resistor R16 and the resistor R17 are connected in series with the capacitor C7; the two ends of the capacitor C7 are respectively connected with the output of the low-pass filter, the The input end of the second-stage amplifying circuit is connected; the output end of the second-stage amplifying circuit is connected with the signal acquisition card, and the signal output end of the signal acquisition card is connected with the input end of the upper computer. The electroporation signal output line is connected with the analog output module of the signal acquisition card.

Further, there are multiple working electrodes and reference electrodes, the multiple working electrodes and reference electrodes are distributed symmetrically around the center, and the distances between all working electrodes and reference electrodes are not greater than 1 mm.

Further, the hollow platinum nanotube array sensor is provided with pin headers, the PCB substrate is provided with corresponding jacks, the pin headers are connected with the corresponding working electrodes and reference electrodes, and the jacks are connected with the sensor electrical signal conditioning circuit.

Further, the working electrode and the reference electrode are prepared by the following steps:

(1) Electrode patterns of working electrode and reference electrode obtained by photolithography: using PET film with a pore diameter of 450nm as an insulating substrate, the electrode end of the working electrode is prepared by photolithography, with a length of 6.5mm, a width of 20-100μm, and a length of the connecting end of 5mm. , the width is 2mm; the electrode end of the reference electrode is 4.5mm long, the width is 20-100μm, the length of the connecting end is 8mm, and the electrode pattern is 2mm wide to obtain a patterned PET film.

(2) Electrode plating: magnetron sputtering 30nm gold or platinum on the patterned PET film obtained in step (1), removing the gold or platinum outside the electrode arrangement pattern to obtain a conductive PET film.

(3) Preparation of hollow platinum nanotube arrays: the metal surface of the PET film is used as the working electrode, the Ag/AgCl electrode is used as the reference electrode, and the platinum wire is used as the counter electrode. Electrolyte was electrodeposited for 200 s under constant current working mode, and hollow platinum nanotube-like structures were formed on the pore walls of the conductive PET film. Part of the PET on the unsputtered metal surface on the PET film is then etched away by _O2 plasma, exposing the electrode arrangement hollow platinum nanotube array with a diameter of 0.4-1 μm and a length of 0.5-2 μm.

Further, the first-stage amplifying circuit further includes a capacitor, and the capacitor is arranged between the inverting input terminal and the output terminal of the operational amplifier in the proportional amplifier of the same phase. The capacitor can be compensated with hysteresis to prevent self-oscillation of the same-phase proportional amplifier. The low-pass filter is a Sallen-Key low-pass filter composed of a low-noise operational amplifier U2A, a capacitor C5 of 1nF to 100nF, a capacitor C6, and a resistor R12, a resistor R13, a resistor R15, a resistor R20, and a resistor R21 with a resistance value of 1k to 50k. filter.

The beneficial effect of the present invention is that the present invention can record the intracellular electrical signal of the cardiomyocyte while ensuring that the basic activity of the cardiomyocyte is not affected, thereby realizing automatic long-term recording of the intracellular electrical signal of the cardiomyocyte.

Description of drawings

Fig. 1 is a scanning electron microscope characterization diagram of hollow platinum nanotubes;

Fig. 2 is a schematic diagram of a hollow platinum nanotube array sensor;

Figure 3 is a schematic diagram of the hollow platinum nanotube array recording and regulating cardiomyocytes;

4 is a block diagram of a sensor electrical signal conditioning and electroporation circuit;

Fig. 5 is a block diagram of the integrated recording and regulation system of myocardial electroporation and intracellular electrical signals of hollow platinum nanotube arrays;

Figure 6 is a schematic diagram of the sensor electrical signal conditioning and electroporation circuit;

Fig. 7 is the program flow chart of the upper computer control software;

Fig. 8 is the main interface of the host computer of the integrated recording and regulation system of myocardial electroporation and intracellular electrical signal of hollow platinum nanotube array;

FIG. 9 is a graph showing the result of real-time data acquisition of the integrated recording and regulation system of myocardial electroporation and intracellular electrical signals of hollow platinum nanotube arrays.

In the figure, working electrode 1, reference electrode 2, PCB board 3, needle row 4, cell culture chamber 5, PET film 6, hollow platinum nanotube array 7, cell 8, needle row jack 9, hollow platinum nanotube array sensor 10. Sensor electrical signal conditioning circuit 11, conditioned sensor electrical signal output terminal 12, power supply interface 13, conditioned sensor electrical signal output terminal 14, PCB substrate 15, acquisition card No. 1 16, acquisition card No. 2 No. 17, Electroporation signal output line 18.

Detailed ways

The detection principle of the intracellular electrical signal of the cardiomyocytes used is described in detail below.

Cardiomyocytes are the basic constituent cells of the heart. They have electrical excitability and automatically generate action potential signals. The generation of action potentials is caused by the inflow and outflow of sodium ions, potassium ions and calcium ions in the corresponding ion channels on the cell membrane. The action potential will be conducted to the outside of the cell through the cardiomyocyte membrane, thereby forming the extracellular field signal of the cardiomyocyte, and these extracellular signals can be recorded by ordinary microelectrode arrays. In order to record the intracellular electrical signal, a nano-electrode array electrode needs to be used, and electroporation is performed through the analog output module of the data acquisition card to discharge the hollow platinum nanotube array tip of the micro-electrode 1 on the sensor 10, thereby causing tiny nano-cracks in the cell membrane. , so that the nano-electrode array can record the intracellular electrical signal, and the intracellular electrical signal is amplified and filtered by the high input impedance low noise amplifier, and then collected and analyzed by the analog input module of the data acquisition card.

The function of the present invention is further described below in conjunction with examples and accompanying drawings, which can better represent the purpose and effect of the present invention.

As shown in FIG. 4 , the integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes of the present invention includes a host computer, a PCB substrate 15 , a hollow platinum nanotube array sensor 10 fixed on the PCB substrate 15 , a sensor electrical Signal conditioning circuit 11, electroporation signal output line 18 and signal acquisition card.

As shown in FIG. 2 , the hollow platinum nanotube array sensor 10 includes a PCB board 3 , a working electrode 1 and a reference electrode 2 . The electrode end of the working electrode 1 is 6.5 mm long, 20-100 μm wide, and the connecting end is 5 mm long. The width is 2 mm; the electrode end of the reference electrode 2 is 4.5 mm long and 20-100 μm wide, and the connecting end is 8 mm long and 2 mm wide. The working electrode 1 is connected to the input end of the sensor electrical signal conditioning circuit 11; the reference electrode 2 is grounded. The working electrode 1 and the reference electrode 2 are fixed on the PCB board 3 through connecting ends, and the connecting ends are connected to the pads on the PCB board 3 through conductive silver paste. A cylinder is fixed on the working electrode 1 and the reference electrode 2 as a cell culture chamber 5 .

Preferably, there are multiple working electrodes 1 and reference electrodes 2 on the hollow platinum nanotube array sensor 10 , and the multiple working electrodes 1 and reference electrodes 2 are distributed symmetrically in the center, which is convenient for bonding with the cylindrical cell culture chamber 5 . The distance between all working electrodes 1 and reference electrodes 2 is not greater than 1mm, so as to avoid the cell impedance is too large and the circuit cannot measure the signal. FIG. 2 is a schematic structural diagram of a hollow platinum nanotube array sensor 10 with 16 working electrodes 1 and 4 reference electrodes 2 .

Both the working electrode 1 and the reference electrode 2 are composed of a PET film 6 and a hollow platinum nanotube array 7 with a diameter of 0.4-1 μm and a length of 0.5-2 μm grown on the PET film 6 . As shown in FIG. 3 , the electrode with this three-dimensional structure has a better coupling effect with the cell membrane of the cell 8 on its surface, which is beneficial to improve the detection sensitivity of the signal.

In this embodiment, a polyethylene terephthalate (PET) polymer film-PET film 6 with a pore size of 0.4-1 μm is used as a template to prepare a hollow platinum nanotube electrode array 7 . First, using photolithography technology, spin-coat a layer of RZJ-390PG positive photoresist on the PET film 6, and irradiate the surface with ultraviolet light through the mask for exposure. After soaking in the developer, the exposed part of the photoresist is exposed. After being removed, the patterned PET film 6 is obtained. Then, the developed PET film 6 is subjected to magnetron sputtering of 30 nm gold or platinum, and then acetone is used to dissolve the remaining photoresist, so that the conductive PET film 6 can be obtained. Then, electrochemical deposition technology is used, the metal-plated surface of PET film 6 is used as the working electrode, the Ag/AgCl electrode is used as the reference electrode, and the platinum wire is used as the counter electrode to form a three-electrode system. The PET film 6 is immersed in the electrolyte, and the electrolytic The composition of the liquid was 1 wt % chloroplatinic acid and 0.5 M hydrochloric acid. Electrodeposition was performed for 200 s in a constant current/constant voltage working mode, and a hollow platinum nanotube-like structure was formed on the pore wall of the PET film 6 . Subsequently, the upper surface of the unsputtered metal surface of the PET film 6 is etched by O ₂ plasma to expose the hollow platinum nanotube array 7 structure with a diameter of 0.4-1 μm and a length of 0.5-2 μm.

The hollow platinum nanotube electrode array 7 is facing upward, and the bottom surface is connected to the PCB board 3 with conductive silver paste. Then, the upper pin 4 is welded, and finally used on the top of the electrode to bond cells with uncured PDMS. The culture chamber 5 (about 1.4 cm in diameter) was placed at 80° C. for 2 hours to solidify the PDMS.

The signal conditioning circuit is composed of a first-stage amplifying circuit, an RC blocking circuit, a low-pass filter, and a second-stage amplifying circuit. As shown in Figure 6, the first-stage amplifying circuit includes a first operational amplifier U1A, a device A capacitor C8 between the reverse input terminal and the output terminal of the first operational amplifier U1A, a resistor R19 connected in parallel with the capacitor C8, a resistor R23 whose one end is connected to the reverse input terminal of the first operational amplifier U1A, and one end connected to the first operational amplifier U1A. Put the resistor R11 connected to the positive input end of U1A, the other end of the resistor R11 is connected to the working electrode, the other end of the resistor R23 is connected to the ground, and the reference electrode 2 is connected to the ground. The resistance values of the resistors R11, R19 and R23 are all between 1kΩ and 200kΩ, and the capacitance value of the feedback capacitor C8 is between 22pF and 1nF. Its function is to compensate the phase and prevent the self-oscillation of U1A. The low-pass filter is a low-noise filter. Operational amplifier U2A, capacitor C5 of 1nF～100nF, capacitor C6, resistor R12, resistor R13, resistor R15, resistor R20 and resistor R21 with resistance value of 1k～50k form a Sallen-Key structure low-pass filter, low noise op amp U2A The positive input terminal of the R13 is connected to capacitor C6 and resistor R13, the other end of C6 is grounded, the other end of resistor R13 is connected to resistor R12 and capacitor C5 respectively, R12 is connected to the output terminal of resistor R15 and U1A, the other end of R15 is grounded, and the other end of capacitor C5 is connected To the output end of U2A, the output end of U2A is connected in series with R21 and R20 to the ground to form a feedback loop, and the inverting input end of U2A is connected to R20 and R21 at the same time; the RC blocking circuit is composed of a 10μF capacitor C7, a 30KΩ resistor R16, and a resistor R17. R16, resistor R17 is connected in series with capacitor C7, and one end of capacitor C7 is connected to the output end of the second operational amplifier U2A; the second stage amplifier circuit consists of precision operational amplifier U3A, one end of which is connected to the output end of precision operational amplifier U3A. Resistor R5 It is composed of resistor R14, resistor R18 and resistor R22 of 1k to 49.9k. Resistor R18 is set between the reverse input terminal and output terminal of precision operational amplifier U3A, one end of resistor R22 is connected to the reverse input terminal of precision operational amplifier U3A, and the other end is grounded; one end of resistor R14 is connected to the positive terminal of precision operational amplifier U3A. It is connected to the input end, and the other end is connected to the capacitor C7. One end of the resistor R5 is grounded, and the other end is connected to the output end of the precision operational amplifier U3A; the output end of the precision operational amplifier U3A is connected to the signal acquisition card, the signal output end of the signal acquisition card is connected to the input end of the host computer, and the same signal conditioning The other channel of the circuit is the same as the previously described circuit. The electroporation signal output line 18 is connected to the analog output channel of the signal acquisition card.

The electronic components related to the above circuits are welded on the sensor electrical signal conditioning circuit module. If dual op amps are used, each channel contains two channels, then another 7 channels of the same sensor electrical signal conditioning circuit 11 are required on the board, forming 16. The channel sensor electrical signal conditioning circuit 11 corresponds to the 16 working electrodes in Figure 2. The eight-channel signal conditioning circuit on the right side of the sensor electrical signal conditioning and electroporation circuit module is connected to the signal output terminal 12. The four-way eight-channel signal conditioning circuit of the perforated circuit module is connected to the signal output end 13, the power supply interface 13 is connected to an external ±5V voltage, and then the signal output end 12 is connected to the data acquisition card 16 by a silicone cable, and the signal output end 14 is made of silica gel The cable is connected with the data acquisition 17, and the data acquisition card 16 and the data acquisition card 17 are connected to the USB hub and then interconnected with the host computer. The host computer is used to control the electroporation signal output, display and record the data obtained by the hollow platinum nanotube array sensor 10 tested.

Preferably, the working electrode 1 and the reference electrode 2 and the sensor electrical signal conditioning circuit 11 are structurally connected to the jack 9 through the pin header 4, the hollow platinum nanotube array sensor 10 is provided with the pin header 4, and the PCB substrate 15 is provided with a corresponding pin header 4. The jack 9, the pin header 4 is connected with the corresponding working electrode 1 and the reference electrode 2, and the jack 9 is connected with the sensor electrical signal conditioning circuit 11.

As shown in FIG. 5 , the working process of the present invention is as follows: with the hollow platinum nanotube array 7 facing upward, insert the pin header 4 of the hollow platinum nanotube array sensor 10 into the corresponding pin header slot 9, The mouse cardiomyocytes 8 are cultured on the upper side, the electroporation signal output line 18 is inserted into the culture chamber 5, the ±5V power supply is turned on, the sensor electrical signal conditioning circuit 18 starts to work, click on the host computer to start the acquisition, and the PC computer will control the data acquisition The card outputs an electroporation pulse signal. The electroporation time is less than 100ms, and the electroporation voltage is generally less than 10mV. The mouse cardiomyocytes in the culture chamber will generate intracellular electrical signals, and the intracellular electrical signals are transmitted to the sensor via the connected pin 4. At the input end of the signal conditioning circuit 11, the electrical signal is first amplified by a proportional amplifier of the same phase, and then low-pass filtered to reduce high-frequency noise, the RC blocking circuit filters out the baseline, and then the second-stage amplification is finally transmitted to the data acquisition card 16 , 17 After collection, it is transmitted to the host computer for display and recording.

Preferably, as shown in Figures 7-8, the workflow of the host computer of the integrated recording and regulation system for the electroporation of the hollow platinum nanotube array and the intracellular electrical signal is as follows: open the program to enter the interface, select whether to record data, and click to start the acquisition , the host computer will record the start time of the experiment according to the current system time, and create a new TDMS record file in the current directory according to the starting time as the naming format. One thread of the host computer will send the collected data into the buffer queue, and the other thread will extract the data from the buffer queue according to the first-in-first-out rule, and then restore the magnification and send it to the waveform chart for display and TDMS file recording. The use of synchronous queues to collect and display data effectively avoids the situation of data loss caused by not being able to collect data at the same time when processing data. When a certain amount of data is collected, the user can click the start collection button again. At this time, the button changes to stop, indicating that the current state is stopped.

The application cases of the present invention are given below.

The integrated recording and regulation system of cardiac electroporation and intracellular electrical signals of hollow platinum nanotube arrays is mainly used for the detection of intracellular electrical signals in cardiomyocytes. In the experiment, the cardiomyocytes and their culture medium were firstly added to the culture chamber 5, the hollow platinum nanotube array sensor 10 was inserted into the slot 9, and the sensor electrical signal conditioning circuit 11 and the data acquisition card were adjusted using the silicone cable and data. , connect the USB hub to the host computer, insert the electroporation signal output line 18 into the culture chamber 5, turn on the ±5V power supply switch, open the host computer software, enter the main interface as shown in Figure 8, check the record data, click to open , wait for about 1 second, the data will be displayed on the waveform chart as shown in Figure 9, which contains 16 channels of data.

The invention adopts the conductive metal nano-needle tube array, develops and applies an automatic electroporation and intracellular electrical signal recording system, and realizes the safe and stable long-term recording of the intracellular electrical signal of myocardial cells.

Claims (5)

1. a cardiomyocyte electroporation measures the integrated recording regulation system of intracellular electrical signal, it is characterized in that, comprise host computer, PCB substrate (15), be fixed on the hollow platinum nanotube array sensor (15) on PCB substrate (15). 10), a sensor electrical signal conditioning circuit (11), an electroporation signal output line (18) and a signal acquisition card; The hollow platinum nanotube array sensor (10) comprises a PCB board (3), a working electrode (1) and a reference electrode (2). The electrode end of the working electrode (1) is 6.5 mm long and 20-100 μm wide, and the connecting end The length is 5mm and the width is 2mm; the electrode end of the reference electrode (2) is 4.5mm long and 20-100μm wide, and the connecting end is 8mm long and 2mm wide; the working electrode (1) and the sensor electrical signal conditioning circuit (11) The reference electrode (2) is grounded; the working electrode (1) and the reference electrode (2) are fixed on the PCB board (3) through the connecting end; a cylinder is fixed on the working electrode (1) and the reference electrode (2) body as a cell culture chamber (5); The working electrode (1) and the reference electrode (2) are both composed of a PET film (6) and a hollow platinum nanotube array (7) with a diameter of 0.4-1 μm and a length of 0.5-2 μm grown on the PET film (6); The host computer is used to control the output of electroporation signals, display and record the data obtained by the hollow platinum nanotube array sensor (10) test; The sensor electrical signal conditioning circuit (11) is composed of a first-stage amplifying circuit, a low-pass filter, an RC blocking circuit and a second-stage amplifying circuit, which are connected in sequence, and the first-stage amplifying circuit and the second-stage amplifying circuit are both The same-phase proportional amplifier; the RC blocking circuit is composed of a 10μF capacitor C7 and a resistor R16 and a resistor R17 with a resistance value of 30kΩ. The resistor R16 and the resistor R17 are connected in series with the capacitor C7; the two ends of the capacitor C7 are respectively connected with the output of the low-pass filter. The output end of the second-stage amplifying circuit is connected to the signal acquisition card, and the signal output end of the signal acquisition card is connected to the input end of the upper computer; the electroporation signal output line (18) is connected to the The analog output module of the signal acquisition card is connected. 2. The integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes according to claim 1, characterized in that, the working electrode (1) and the reference electrode (2) are multiple, and the multiple working electrodes (1) and the reference electrode (2) are multiple. The electrodes (1) and the reference electrodes (2) are distributed symmetrically in the center, and the distances between all the working electrodes (1) and the reference electrodes (2) are not greater than 1 mm. 3. The integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes according to claim 1, is characterized in that, the hollow platinum nanotube array sensor (10) is provided with needles (4), the PCB substrate (15) is provided with a corresponding jack (9), the pin header (4) is connected with the corresponding working electrode (1) and the reference electrode (2), and the jack (9) is connected with the sensor electrical signal conditioning circuit (11) connected. 4. The integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes according to claim 1, wherein the working electrode (1) and the reference electrode (2) are obtained by the following steps: (1) The patterns of the working electrode (1) and the reference electrode (2) are obtained by photolithography: using a PET film with an aperture of 450 nm as an insulating substrate, the electrode end of the working electrode (1) is prepared by photolithography with a length of 6.5 mm and a width of 20 mm. -100 μm, the length of the connecting end is 5 mm, and the width is 2 mm; the electrode end of the reference electrode (2) is 4.5 mm long, the width is 20-100 μm, the length of the connecting end is 8 mm, and the width is 2 mm. PET film; (2) Electrode plating: magnetron sputtering 30nm gold or platinum on the patterned PET film obtained in step (1), remove the gold or platinum outside the electrode arrangement pattern to obtain a conductive PET film; (3) Preparation of hollow platinum nanotube arrays: the metal surface of the PET film is used as the working electrode, the Ag/AgCl electrode is used as the reference electrode, and the platinum wire is used as the counter electrode. Electrolyte was electrodeposited for 200s under constant current working mode to form hollow platinum nanotube-like structures on the pore walls of the conductive PET film; then _O2 plasma was used to etch away part of the PET on the unsputtered metal surface of the PET film, exposing the diameter Electrodes with a length of 0.4-1 μm and a length of 0.5-2 μm are arranged with hollow platinum nanotube arrays. 5. The integrated recording and regulating system for measuring intracellular electrical signals by electroporation of cardiomyocytes according to claim 1, wherein the first-stage amplifying circuit further comprises a capacitor, and the capacitor is arranged on the inverter of the operational amplifier in the same-phase proportional amplifier. To between the input end and the output end; the low-pass filter is composed of low-noise operational amplifier U2A, capacitor C5 of 1nF to 100nF, capacitor C6 and resistor R12, resistor R13, resistor R15, resistor R20 and A Sallen-Key low-pass filter composed of resistor R21.

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