Myocardial cell intracellular action potential recording system based on double-phase pulse electroporation

Abstract

The invention discloses a myocardial cell intracellular action potential recording system based on double-phase pulse electroporation, and belongs to the technical field of electrophysiological recording, and the system comprises a double-phase pulse electroporation module which is connected with a microcontroller and a myocardial cell sensor, and is used for receiving a pulse signal sent by the microcontroller, converting the pulse signal into a double-phase pulse signal, and transmitting the double-phase pulse signal to the myocardial cell sensor; issuing to myocardial cells; the myocardial cell sensor is used for acquiring intracellular electric signals, transmitting the intracellular electric signals to the data acquisition card module through the first-stage amplification circuit and the second-stage amplification circuit which are connected in sequence, and outputting the intracellular electric signals to the microcontroller through the analog-to-digital converter, so that the intracellular action potential recording of myocardial cells is completed. According to the application, the biphase pulse signal is used as the electroporation stimulation signal, so that the damage to the cell membrane can be effectively reduced, the integrity and physiological activity of the cell can be maintained, and the electrical signal change of the cell before and after stimulation can be better recorded, thereby better recording the intracellular action potential and reflecting the electrophysiological characteristics of the cell.

Description

Technical Field

[0001] The present application relates to the technical field of electrophysiological recording, and in particular to a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation. Background Art

[0002] Electrophysiological recording technology has been widely used in neuroscience, physiology, and clinical medicine research, becoming an important tool for studying the cardiovascular system and other physiological processes. By capturing electrical activity within a living organism, this technology can reveal key information such as its temporal characteristics, frequency characteristics, and spatial distribution, thereby helping researchers gain a deeper understanding of systemic functions and the pathological mechanisms of related diseases.

[0003] In related technologies, electroporation, as an efficient and stable cell perforation technology, is combined with microelectrode arrays to create nanoscale pores in the cell membrane by forming a localized electric field on the cell membrane, thereby achieving high-throughput recording of intracellular action potentials. However, in actual applications, it has been found that the electroporation technology in related technologies usually uses a single-phase voltage pulse signal, which often causes irreversible damage to the cell membrane, thereby affecting the normal physiological function and signal conduction of the cell. It is also easy to cause abnormal fluctuations in the intracellular calcium ion concentration, interfering with the cell's metabolic activity, thereby affecting the accuracy of intracellular action potential recording.

[0004] In summary, the technical problems existing in the relevant technologies need to be improved. Summary of the Invention

[0005] The embodiment of the present application provides a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation, which can effectively reduce damage to the cell membrane, maintain the integrity and physiological activity of the cells, better record the changes in the electrical signals of the cells before and after stimulation, thereby better recording the intracellular action potential and reflecting the electrophysiological characteristics of the cells.

[0006] On the one hand, an embodiment of the present application provides a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation, the system comprising: a biphasic pulse electroporation module, a cardiomyocyte sensor, a microcontroller, a primary amplification circuit, a secondary amplification circuit, a data acquisition card module, and a power supply module;

[0007] The input end of the biphasic pulse electroporation module is connected to the microcontroller, and the output end of the biphasic pulse electroporation module is connected to the myocardial cell sensor. The biphasic pulse electroporation module is used to receive the pulse signal sent by the microcontroller and convert the pulse signal into a biphasic pulse signal, thereby delivering the biphasic pulse signal to the myocardial cells through the myocardial cell sensor;

[0008] The myocardial cell sensor is used to collect the intracellular electrical signals of myocardial cells, and transmit the intracellular electrical signals to the input end of the data acquisition card module through the sequentially connected first-stage amplifier circuit and the second-stage amplifier circuit;

[0009] The output end of the data acquisition card module is connected to the microcontroller. The data acquisition card module is used to output the received intracellular electrical signals to the microcontroller through an analog-to-digital converter, thereby completing the recording of the intracellular action potential of myocardial cells;

[0010] The power supply module is used to supply power to the first-stage amplifier circuit, the second-stage amplifier circuit and the biphasic pulse electroporation module.

[0011] Optionally, the first-stage amplifier circuit includes: a first resistor, a second resistor, a third resistor, a fourth resistor, a first capacitor, a second capacitor and a first operational amplifier;

[0012] One end of the first capacitor is connected to the input end of the first-stage amplifier circuit;

[0013] One end of the first resistor and one end of the second resistor are connected to the other end of the first capacitor;

[0014] The other end of the first resistor is grounded; the other end of the second resistor is connected to the non-inverting input end of the first operational amplifier;

[0015] One end of the third resistor is connected to the inverting input end of the first operational amplifier; the other end of the third resistor is connected to the output end of the first operational amplifier;

[0016] One end of the fourth resistor is connected to the inverting input end of the first operational amplifier; the other end of the fourth resistor is grounded;

[0017] One end of the second capacitor is connected to the inverting input end of the first operational amplifier; the other end of the second capacitor is connected to the output end of the first operational amplifier;

[0018] The output end of the first operational amplifier is connected to the output end of the first-stage amplifier circuit.

[0019] Optionally, the second-stage amplifier circuit includes a fifth resistor, a sixth resistor, a seventh resistor, an eighth resistor, a ninth resistor, a tenth resistor, a third capacitor, a fourth capacitor, a fifth capacitor, a sixth capacitor, a second operational amplifier and a third operational amplifier;

[0020] One end of the third capacitor is connected to the input end of the second-stage amplifier circuit;

[0021] One end of the fifth resistor and one end of the sixth resistor are connected to the other end of the third capacitor;

[0022] The other end of the fifth resistor is grounded;

[0023] The other end of the sixth resistor and one end of the seventh resistor are connected to one end of the fourth capacitor;

[0024] The other end of the fourth capacitor is connected to the output terminal of the second operational amplifier;

[0025] The other end of the seventh resistor and one end of the fifth capacitor are connected to the non-inverting input terminal of the second operational amplifier;

[0026] The other end of the fifth capacitor is grounded; the inverting input terminal of the second operational amplifier is connected to the output terminal of the second operational amplifier;

[0027] One end of the eighth resistor is connected to the output terminal of the second operational amplifier; the other end of the eighth resistor is connected to the non-inverting input terminal of the third operational amplifier;

[0028] One end of the ninth resistor is connected to the inverting input terminal of the third operational amplifier; the other end of the ninth resistor is connected to the output terminal of the third operational amplifier;

[0029] One end of the tenth resistor is connected to the inverting input terminal of the third operational amplifier; the other end of the tenth resistor is grounded;

[0030] One end of the sixth capacitor is connected to the inverting input terminal of the third operational amplifier; the other end of the sixth capacitor is connected to the output terminal of the third operational amplifier;

[0031] The output terminal of the third operational amplifier is connected to the output terminal of the secondary amplification circuit.

[0032] Optionally, the microcontroller is an STM32 microcontroller.

[0033] Optionally, the biphasic pulse electroporation module includes a fourth operational amplifier, a fifth operational amplifier, a sixth operational amplifier, an analog switch, and a multiplexer;

[0034] The DAC pin of the STM32 microcontroller is connected to the input terminal of the fourth operational amplifier, for converting the pulse signal sent by the DAC pin into a positive-phase pulse signal;

[0035] The output terminal of the fourth operational amplifier is connected to the analog switch, for transmitting the positive-phase pulse signal to the analog switch;

[0036] The output terminal of the fourth operational amplifier is also connected to the input terminal of the fifth operational amplifier, and is used to convert the positive-phase pulse signal into a negative-phase pulse signal through the fifth operational amplifier;

[0037] The output terminal of the fifth operational amplifier is connected to the analog switch, and is used to transmit the negative-phase pulse signal to the analog switch;

[0038] The PA5 pin and PA8 pin of the stm32 microcontroller are connected to the analog switch, and are used to regulate the pulse widths of the positive-phase pulse signal and the negative-phase pulse signal to generate a biphasic pulse signal;

[0039] The output terminal of the analog switch is connected to the multiplexer through the sixth operational amplifier;

[0040] The IO port pin of the stm32 microcontroller is connected to the multiplexer, and is used to enable and control the biphasic pulse electroporation module to send the biphasic pulse signal to cardiomyocytes through the cardiomyocyte sensor.

[0041] Optionally, the power supply module includes a power adapter and a power output module;

[0042] The power adapter is used to convert the commercial power into a DC power supply with a first rated value;

[0043] The power output module is used to convert the DC power supply with the first rated value into a DC power supply with a second rated value, a DC power supply with a third rated value, and a DC power supply with a fourth rated value;

[0044] Wherein, the DC power supply with the second rated value and the DC power supply with the third rated value supply power to the first-stage amplification circuit; the DC power supply with the second rated value and the DC power supply with the third rated value supply power to the second-stage amplification circuit; the DC power supply with the fourth rated value supplies power to the biphasic pulse electroporation module.

[0045] Optionally, the DC power supply with the first rated value is a +12V DC power supply; the DC power supply with the second rated value is a +5V DC power supply; the DC power supply with the third rated value is a -5V DC power supply; the DC power supply with the fourth rated value is a +18V DC power supply.

[0046] On the other hand, an embodiment of the present application provides a method for recording intracellular action potential of cardiomyocytes based on biphasic pulse electroporation, and the method includes the following steps:

[0047] Sending a pulse signal; wherein, the pulse signal is received by the biphasic pulse electroporation module and converted into a biphasic pulse signal, so as to send the biphasic pulse signal to cardiomyocytes through the cardiomyocyte sensor;

[0048] Receive the intracellular electrical signal of the cardiomyocytes to complete the recording of the intracellular action potential of the cardiomyocytes; wherein, the intracellular electrical signal is collected by the cardiomyocyte sensor and transmitted through a first-stage amplifier circuit, a second-stage amplifier circuit, and a data acquisition card module connected in sequence.

[0049] On the other hand, an embodiment of the present application provides an electronic device, which includes a memory and a processor. The memory stores a computer program, and when the processor executes the computer program, the above-mentioned method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation is implemented.

[0050] On the other hand, an embodiment of the present application provides a computer-readable storage medium, which stores a computer program. When the computer program is executed by a processor, the above-mentioned method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation is implemented.

[0051] By using a biphasic pulse signal as the electroporation stimulation signal in the embodiment of the present application, the damage to the cell membrane can be effectively reduced, the integrity and physiological activity of the cells can be maintained, the change of the electrical signal of the cells before and after stimulation can be better recorded, so as to better record the intracellular action potential and reflect the electrophysiological characteristics of the cells. Description of the Drawings

[0052] Figure 1 is a schematic structural diagram of a system for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation provided by an embodiment of the present application;

[0053] Figure 2 is a schematic structural diagram of a first-stage amplifier circuit provided by an embodiment of the present application;

[0054] Figure 3 is a schematic structural diagram of a second-stage amplifier circuit provided by an embodiment of the present application;

[0055] Figure 4 is a schematic structural diagram of a biphasic pulse electroporation module provided by an embodiment of the present application;

[0056] Figure 5 is a schematic structural diagram of a power supply module provided by an embodiment of the present application;

[0057] Figure 6 is a schematic flow diagram of a method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation provided by an embodiment of the present application

[0058] Figure 7 is a schematic hardware structure diagram of an electronic device provided by an embodiment of the present application. Detailed Embodiments

[0059] In order to make the objectives, technical solutions, and advantages of this application clearer and more understandable, the following further details this application in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain this application and are not used to limit this application. When the following description involves the accompanying drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The implementation manners described in the following exemplary embodiments do not represent all implementation manners consistent with the embodiments of this application. They are only examples of devices and methods that are consistent with some aspects of the embodiments of this application as detailed in the appended claims.

[0060] It can be understood that the terms "first", "second", etc. used in this application can be used in this article to describe various concepts, but unless otherwise specified, these concepts are not limited by these terms. These terms are only used to distinguish one concept from another. For example, without departing from the scope of the embodiments of this application, the first information can also be referred to as the second information. Similarly, the second information can also be referred to as the first information. Depending on the context, the words "if", "when" as used herein can be interpreted as "when...", "while...", or "in response to determining".

[0061] The terms "at least one", "multiple", "each", "any one", etc. used in this application, at least one includes one, two, or more than two, multiple includes two or more than two, each refers to each of the corresponding multiple, and any one refers to any one of the multiple.

[0062] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which this application belongs. The terms used herein are only for the purpose of describing the embodiments of this application and are not intended to limit this application.

[0063] Electrophysiological recording technology has been widely used in neuroscience, physiology, and clinical medical research and has become an important means for studying the cardiovascular system and other physiological processes. By capturing the electrical activities in living organisms, this technology can reveal key information such as the temporal characteristics, frequency characteristics, and spatial distribution of electrical activities, thereby helping researchers deeply understand the system functions and the pathological mechanisms of related diseases.

[0064] In related technologies, electroporation, as an efficient and stable cell perforation technology, is combined with microelectrode arrays to create nanoscale pores in the cell membrane by forming a localized electric field on the cell membrane, thereby achieving high-throughput recording of intracellular action potentials. However, in actual applications, it has been found that the electroporation technology in related technologies usually uses a single-phase voltage pulse signal, which often causes irreversible damage to the cell membrane, thereby affecting the normal physiological function and signal conduction of the cell. It is also easy to cause abnormal fluctuations in the intracellular calcium ion concentration, interfering with the cell's metabolic activity, thereby affecting the accuracy of intracellular action potential recording.

[0065] In view of this, an embodiment of the present application provides a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation. By adopting a biphasic pulse signal as the electroporation stimulation signal, it can effectively reduce damage to the cell membrane, maintain the integrity and physiological activity of the cell, and better record the changes in the electrical signals of the cell before and after stimulation, thereby better recording the intracellular action potential and reflecting the electrophysiological characteristics of the cell.

[0066] The specific implementation of the embodiment of the present application is described in detail below with reference to the accompanying drawings. First, a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation provided in the embodiment of the present application is described with reference to the accompanying drawings.

[0067] Please refer to Figure 1 , Figure 1 Schematic diagram of a cardiomyocyte intracellular action potential recording system based on biphasic pulse electroporation provided in an embodiment of the present application, the system comprising: a biphasic pulse electroporation module, a cardiomyocyte sensor, a microcontroller, a primary amplification circuit, a secondary amplification circuit, a data acquisition card module, and a power supply module;

[0068] The input end of the biphasic pulse electroporation module is connected to the microcontroller, and the output end of the biphasic pulse electroporation module is connected to the myocardial cell sensor. The biphasic pulse electroporation module is used to receive the pulse signal sent by the microcontroller and convert the pulse signal into a biphasic pulse signal, thereby delivering the biphasic pulse signal to the myocardial cells through the myocardial cell sensor;

[0069] The myocardial cell sensor is used to collect intracellular electrical signals of myocardial cells, and transmit the intracellular electrical signals to the input end of the data acquisition card module through the first-stage amplification circuit and the second-stage amplification circuit connected in sequence;

[0070] The output end of the data acquisition card module is connected to the microcontroller, and the data acquisition card module is used to output the received intracellular electrical signal to the microcontroller through an analog-to-digital converter, thereby completing the recording of intracellular action potentials of myocardial cells;

[0071] The power supply module is used to supply power to the first-stage amplification circuit, the second-stage amplification circuit, and the biphasic pulse electroporation module.

[0072] In the embodiment of the present application, the intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation mainly includes a biphasic pulse electroporation module, a cardiomyocyte sensor, a microcontroller, a first-stage amplification circuit, a second-stage amplification circuit, a data acquisition card module, and a power supply module.

[0073] Among them, the microcontroller is connected to the input end of the biphasic pulse electroporation module, and can send pulse signals to the biphasic pulse electroporation module regularly according to specific usage requirements, so that the biphasic pulse electroporation module converts the pulse signals into biphasic pulse signals, and then sends the biphasic pulse signals to cardiomyocytes through the cardiomyocyte sensor.

[0074] Therefore, when the cardiomyocytes are stimulated by the biphasic pulse signals, a localized electric field is formed on the cell membrane of the cardiomyocytes, causing nanoscale pores to be generated on the cell membrane, and the cardiomyocyte sensor can collect the intracellular electrical signals of the changes in the action potential of the cardiomyocytes after being stimulated.

[0075] Furthermore, after the cardiomyocyte sensor collects the intracellular electrical signals of the cardiomyocytes, considering the problem that the signals are vulnerable to noise interference during the transmission process in this application, the intracellular electrical signals are filtered and gain-amplified through the first-stage amplification circuit and the second-stage amplification circuit, and then transmitted to the data acquisition card module. The analog-to-digital converter of the data acquisition card module converts the intracellular electrical signals into digital signals and transmits them to the microcontroller, and the microcontroller completes the recording of the intracellular action potential of the cardiomyocytes. Among them, the microcontroller can display the waveform of the intracellular electrical signals of the cardiomyocytes, and perform data analysis on the intracellular electrical signals, calculate the amplitude of the intracellular electrical signals, so as to record and store the amplitude of the intracellular action potential.

[0076] In practical applications, the cardiomyocyte sensor can select an MEA (Microelectrode Array) sensor. The microelectrode array can simultaneously monitor the electrical signals of multiple cells or different positions of the same cell. By selecting the 32-channel recording mode, the efficiency and comprehensiveness of data acquisition can be significantly improved.

[0077] Exemplarily, each channel in the MEA sensor can be correspondingly configured with a first-stage amplification circuit and a second-stage amplification circuit. Finally, the intracellular electrical signals of the cardiomyocytes in the cardiomyocyte incubator pass through the microelectrode array sensor with 32 channels, and after signal conditioning, they are output to the data acquisition card module. Finally, the data acquisition card module outputs the intracellular electrical signals to the microcontroller, and the microcontroller completes the recording of the intracellular action potential of the cardiomyocytes.

[0078] Optionally, the electrophysiological interaction between the electrode and the cell can also be significantly enhanced by increasing the surface roughness of the electrodes in the microelectrode array or designing a three-dimensional electrode structure, thereby effectively improving the sensitivity and accuracy of signal recording. In addition, an electrophysiological detection chip (such as RHD2000) can be used to replace the amplification and filtering circuit (the first-stage amplification circuit and the second-stage amplification circuit), which can further reduce the system noise and improve the quality of the intracellular electrical signals collected.

[0079] Finally, the intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation is also provided with a power supply module, which supplies power to the first-stage amplification circuit, the second-stage amplification circuit, and the biphasic pulse electroporation module.

[0080] Since this application stimulates cardiomyocytes with biphasic pulse signals and introduces two voltage pulses with opposite polarities in the stimulation pulse, it can effectively reduce the damage to the cell membrane and maintain the integrity and physiological activity of the cells. In addition, it can more clearly distinguish the spontaneous electrical activity and stimulation-induced electrical activity of the cells, improve the signal-to-noise ratio of the intracellular electrical signals, provide richer and more stable information for cell electrophysiological research, and has important application value.

[0081] Specifically, as an optional implementation manner, please refer to Figure 2 , Figure 2 which is a schematic structural diagram of a first-stage amplification circuit provided by an embodiment of this application. The first-stage amplification circuit includes: a first resistor R1, a second resistor R2, a third resistor R3, a fourth resistor R4, a first capacitor C1, a second capacitor C2, and a first operational amplifier U1;

[0082] One end of the first capacitor C1 is connected to the input end of the first-stage amplification circuit;

[0083] One end of the first resistor R1 and one end of the second resistor R2 are connected to the other end of the first capacitor C1;

[0084] The other end of the first resistor R1 is grounded; the other end of the second resistor R2 is connected to the non-inverting input end of the first operational amplifier U1;

[0085] One end of the third resistor R3 is connected to the inverting input end of the first operational amplifier U1; the other end of the third resistor R3 is connected to the output end of the first operational amplifier U1;

[0086] One end of the fourth resistor R4 is connected to the inverting input end of the first operational amplifier U1; the other end of the fourth resistor R4 is grounded;

[0087] One end of the second capacitor C2 is connected to the inverting input terminal of the first operational amplifier U1; the other end of the second capacitor C2 is connected to the output terminal of the first operational amplifier U1;

[0088] The output terminal of the first operational amplifier U1 is connected to the output terminal of the first-stage amplification circuit.

[0089] In the embodiment of the present application, as Figure 2 shown, the input terminal of the first-stage amplification circuit is connected to the myocardial cell sensor, and the intracellular electrical signal collected by the electrodes in the myocardial cell sensor will be transmitted to the first-stage amplification circuit for signal conditioning.

[0090] Further, the input terminal of the first-stage amplification circuit is connected to one end of the first capacitor C1. One end of the first resistor R1 and one end of the second resistor R2 are connected to the other end of the first capacitor C1. The other end of the first resistor R1 is grounded. Thus, the first capacitor C1 and the first resistor R1 form a passive high-pass filter. Exemplarily, the first resistor R1 can be set to 50 MΩ, and the first capacitor C1 can be set to 3.3 nF. The cut-off frequency of the passive high-pass filter is 0.96 Hz.

[0091] Further, the other end of the second resistor R2 is connected to the non-inverting input terminal of the first operational amplifier U1 as the balancing resistor of the first operational amplifier U1.

[0092] One end of the third resistor R3 is connected to the inverting input terminal of the first operational amplifier U1, and the other end of the third resistor R3 is connected to the output terminal of the first operational amplifier U1. One end of the fourth resistor R4 is connected to the inverting input terminal of the first operational amplifier U1, and the other end of the fourth resistor R4 is grounded. Thus, the third resistor R3 and the fourth resistor R4 can determine the amplification factor of the first operational amplifier U1.

[0093] Exemplarily, the first operational amplifier U1 can be selected as OPA2140 to form a non-inverting proportional amplifier. By setting the third resistor R3 to 887 Ω and the fourth resistor R4 to 100 Ω, a 10-fold amplification factor is formed, and the first operational amplifier U1 is powered by a +5V DC power supply and a -5V DC power supply.

[0094] Further, one end of the second capacitor C2 is connected to the inverting input terminal of the first operational amplifier U1, and the other end of the second capacitor C2 is connected to the output terminal of the first operational amplifier U1. The second capacitor C2 can prevent the first operational amplifier U1 from self-oscillating. Exemplarily, the second capacitor C2 can be set to 33 pF.

[0095] Finally, the output terminal of the first operational amplifier U1 is connected to the output terminal of the first-stage amplification circuit, and the amplified and filtered intracellular electrical signal can be output through the output terminal to the input terminal of the second-stage amplification circuit.

[0096] Specifically, as an alternative embodiment, please refer to Figure 3 , Figure 3 which is a schematic structural diagram of a two-stage amplifier circuit provided by an embodiment of the present application. The two-stage amplifier circuit includes a fifth resistor R5, a sixth resistor R6, a seventh resistor R7, an eighth resistor R8, a ninth resistor R9, a tenth resistor R10, a third capacitor C3, a fourth capacitor C4, a fifth capacitor C5, a sixth capacitor C6, a second operational amplifier U2, and a third operational amplifier U3;

[0097] One end of the third capacitor C3 is connected to the input end of the two-stage amplifier circuit;

[0098] One end of the fifth resistor R5 and one end of the sixth resistor R6 are connected to the other end of the third capacitor C3;

[0099] The other end of the fifth resistor R5 is grounded;

[0100] The other end of the sixth resistor R6 and one end of the seventh resistor R7 are connected to one end of the fourth capacitor C4;

[0101] The other end of the fourth capacitor C4 is connected to the output end of the second operational amplifier U2;

[0102] The other end of the seventh resistor R7 and one end of the fifth capacitor C5 are connected to the non-inverting input end of the second operational amplifier U2;

[0103] The other end of the fifth capacitor C5 is grounded; the inverting input end of the second operational amplifier U2 is connected to the output end of the second operational amplifier U2;

[0104] One end of the eighth resistor R8 is connected to the output end of the second operational amplifier U2; the other end of the eighth resistor R8 is connected to the non-inverting input end of the third operational amplifier U3;

[0105] One end of the ninth resistor R9 is connected to the inverting input end of the third operational amplifier U3; the other end of the ninth resistor R9 is connected to the output end of the third operational amplifier U3;

[0106] One end of the tenth resistor Rl0 is connected to the inverting input end of the third operational amplifier U3; the other end of the tenth resistor Rl0 is grounded;

[0107] One end of the sixth capacitor C6 is connected to the inverting input end of the third operational amplifier U3; the other end of the sixth capacitor C6 is connected to the output end of the third operational amplifier U3;

[0108] The output terminal of the third operational amplifier U3 is connected to the output terminal of the second-stage amplification circuit.

[0109] In the embodiment of the present application, as Figure 3 shown, the second-stage amplification circuit mainly includes three parts: high-pass filtering, low-pass filtering, and second-stage amplification.

[0110] Among them, the input terminal of the second-stage amplification circuit is connected to the output terminal of the first-stage amplification circuit, so as to perform further signal modulation processing on the intracellular electrical signal of cardiomyocytes.

[0111] Further, one end of the third capacitor C3 is connected to the input terminal of the second-stage amplification circuit, one end of the fifth resistor R5 and one end of the sixth resistor R6 are connected to the other end of the third capacitor C3, and the other end of the fifth resistor R5 is grounded. Thus, the third capacitor C3 and the fifth resistor R5 form a high-pass filter.

[0112] Exemplarily, the third capacitor C3 can be set to 10 μF, the fifth resistor R5 can be set to 51 KΩ, and the lower cut-off frequency of the high-pass filter is 1 Hz.

[0113] Further, the other end of the sixth resistor R6 and one end of the seventh resistor R7 are connected to one end of the fourth capacitor C4, the other end of the fourth capacitor C4 is connected to the output terminal of the second operational amplifier U2, the other end of the seventh resistor R7 and one end of the fifth capacitor C5 are connected to the non-inverting input terminal of the second operational amplifier U2, the other end of the fifth capacitor C5 is grounded, and the inverting input terminal and the output terminal of the second operational amplifier U2 are connected. Thus, the sixth resistor R6 serves as the balancing resistor of the second operational amplifier U2, and the seventh resistor R7, the fourth capacitor C4, the fifth capacitor C5, and the second operational amplifier U2 form a second-order low-pass Butterworth filter.

[0114] Exemplarily, the sixth resistor R6 can be set to 1.3 KΩ, the seventh resistor R7 can be set to 1.8 KΩ, the fourth capacitor C4 can be set to 20 nF, the fifth capacitor C5 can be set to 10 nF, and the second operational amplifier U2 can adopt the OPA2227 operational amplifier, and the second operational amplifier U2 is powered by a positive 5V DC power supply and a negative 5V DC power supply.

[0115] Further, after high-pass filtering and low-pass filtering are completed, secondary amplification can be performed. The output terminal of the second operational amplifier U2 is connected to one end of the eighth resistor R8, the other end of the eighth resistor R8 is connected to the non-inverting input terminal of the third operational amplifier U3, one end of the ninth resistor R9 is connected to the inverting input terminal of the third operational amplifier U3, the other end of the ninth resistor R9 is connected to the output terminal of the third operational amplifier U3, one end of the tenth resistor R10 is connected to the inverting input terminal of the third operational amplifier U3, and the other end of the tenth resistor R10 is grounded. Thus, the eighth resistor R8 serves as the balancing resistor of the third operational amplifier U3, and the ninth resistor R9 and the tenth resistor R10 can determine the amplification factor of the third operational amplifier U3.

[0116] Exemplarily, the eighth resistor R8 can be set to 100 Ω, the ninth resistor R9 can be set to 5.1 kΩ, and the tenth resistor R10 can be set to 100 Ω, constituting an amplification factor of 50 times. The third operational amplifier U3 can select the OPA2227 operational amplifier and be powered by a +5V DC power supply and a -5V DC power supply for the third operational amplifier U3.

[0117] Further, one end of the sixth capacitor C6 is connected to the inverting input terminal of the third operational amplifier U3, and the other end of the sixth capacitor C6 is connected to the output terminal of the third operational amplifier U3. The sixth capacitor C6 can prevent the third operational amplifier U3 from self-oscillating. Exemplarily, the sixth capacitor C6 can be set to 22 pF.

[0118] Finally, the output terminal of the third operational amplifier U3 is connected to the data acquisition card module. The amplified and filtered intracellular electrical signal can be output to the data acquisition card module through the output terminal, and then through the analog-to-digital converter of the data acquisition card module, the amplified, filtered, and modulated intracellular electrical signal is transmitted to the microcontroller, so as to facilitate subsequent analysis and calculation of the intracellular electrical signal and complete the recording of the intracellular action potential of cardiomyocytes.

[0119] Specifically, as an optional implementation manner, the microcontroller is an stm32 microcontroller.

[0120] In the embodiment of the present application, an stm32 microcontroller can be selected as the microcontroller of the intracellular action potential recording system of cardiomyocytes based on biphasic pulse electroporation. The stm32 microcontroller sends a pulse signal to the biphasic pulse electroporation module, receives the intracellular electrical signal uploaded by the data acquisition card module, and performs data analysis on the intracellular electrical signal to calculate the amplitude of the intracellular electrical signal, thereby completing the recording and storage of the intracellular action potential.

[0121] Specifically, as an optional implementation manner, please refer to Figure 4 , Figure 4It is a schematic structural diagram of a bipolar pulse electroporation module provided by an embodiment of the present application. The bipolar pulse electroporation module includes a fourth operational amplifier, a fifth operational amplifier, a sixth operational amplifier, an analog switch, and a multiplexer;

[0122] The DAC pin of the stm32 microcontroller is connected to the input end of the fourth operational amplifier, and is used to convert the pulse signal sent by the DAC pin into a positive-phase pulse signal;

[0123] The output end of the fourth operational amplifier is connected to the analog switch, and is used to transmit the positive-phase pulse signal to the analog switch;

[0124] The output end of the fourth operational amplifier is also connected to the input end of the fifth operational amplifier, and is used to convert the positive-phase pulse signal into a negative-phase pulse signal through the fifth operational amplifier;

[0125] The output end of the fifth operational amplifier is connected to the analog switch, and is used to transmit the negative-phase pulse signal to the analog switch;

[0126] The PA5 pin and PA8 pin of the stm32 microcontroller are connected to the analog switch, and are used to regulate the pulse widths of the positive-phase pulse signal and the negative-phase pulse signal to generate a bipolar pulse signal;

[0127] The output end of the analog switch passes through the sixth operational amplifier and is connected to the multiplexer;

[0128] The IO port pin of the stm32 microcontroller is connected to the multiplexer, and is used to enable the control of the bipolar pulse electroporation module to issue the bipolar pulse signal to cardiomyocytes through the cardiomyocyte sensor.

[0129] In the embodiment of the present application, as Figure 4 shown, a pulse signal can be generated by using the DAC (digital-to-analog converter) pin of the stm32 microcontroller, and the pulse signal is transmitted to the non-inverting amplifier circuit composed of the fourth operational amplifier to obtain a positive-phase pulse signal.

[0130] Further, the positive-phase pulse signal is transmitted to the analog switch, so as to transmit the positive-phase pulse signal to the analog switch, and is also transmitted to the fifth operational amplifier set as an inverting amplifier to convert the positive-phase pulse signal into a negative-phase pulse signal, and transmit the negative-phase pulse signal to the analog switch.

[0131] Exemplarily, the fourth operational amplifier and the fifth operational amplifier can adopt the OPA2140 operational amplifier, and the fourth operational amplifier and the fifth operational amplifier are powered by a +18V DC power supply and a -18V DC power supply. Among them, the +18V DC power supply can be obtained from the power output module in the power supply module, and the -18V DC power supply can be powered by a commercial power supply.

[0132] Further, the analog switch can select the CD4052B analog switch. By connecting the PA5 pin and the PA8 pin of the stm32 microcontroller to the address terminal of the CD4052B analog switch, connecting the positive-phase pulse signal and the negative-phase pulse signal to the Y-channel input terminal of the CD4052B analog switch, and connecting the X common output terminal of the analog switch to the X-channel input terminal corresponding to the Y-channel input terminal to which the positive-phase pulse signal and the negative-phase pulse signal have been connected, the specific value of the address terminal of the CD4052B analog switch is determined by the signals output from the PA5 pin and the PA8 pin of the stm32 microcontroller, so as to control the connection of the corresponding X and Y channel input terminals, and further enable the Y-channel common output terminal to obtain the bipolar pulse signal composed of the positive-phase pulse signal and the negative-phase pulse signal. Among them, the pulse widths of the positive-phase pulse signal and the negative-phase pulse signal are controlled by the PA5 pin and the PA8 pin.

[0133] Further, the output terminal of the analog switch is connected to the multiplexer through the sixth operational amplifier, and the multiplexer is enabled and controlled by the IO port pins of the stm32 microcontroller, so as to determine whether to issue the bipolar pulse signal to the cardiomyocytes through the cardiomyocyte sensor.

[0134] Exemplarily, the 32 IO port pins on the stm32 microcontroller can be used to enable and control the path switches in the multiplexer one by one, so as to control the switches of 32 paths of the MEA sensor. For example, when the 1st IO port pin on the stm32 microcontroller sends out a high-level signal, the 1st path switch in the multiplexer will be correspondingly enabled and closed, and the bipolar pulse signal will be transmitted to the 1st path of the MEA sensor to issue and stimulate the cardiomyocytes on this path.

[0135] Exemplarily, the sixth operational amplifier can select the OPA455 operational amplifier, the amplification factor can be set to 6 times, and the multiplexer can select the TMUX6212 multiplexer.

[0136] Therefore, a pulse signal is generated by the STM32 microcontroller, and a positive-phase pulse signal and a negative-phase pulse signal are generated through the fourth operational amplifier and the fifth operational amplifier. Then, they are transmitted to the channel input end of the analog switch. The PA5 and PA8 pins of the STM32 microcontroller are used as the address-end control signals of the analog switch. By intermittently connecting the positive-phase pulse signal and the negative-phase pulse signal, the output end of the analog switch will correspondingly output a bipolar pulse signal. The bipolar pulse signal is amplified by the sixth operational amplifier and then input into multiple paths in the multiplexer. Each path of the multiplexer corresponds to a path in the myocardial cell sensor (MEA sensor), and is enabled and controlled by the IO port pins of the STM32 microcontroller. By sending high and low level signals to control the closing and opening of the path, the bipolar pulse electroporation module is enabled to send a bipolar pulse signal to the myocardial cells through the myocardial cell sensor.

[0137] Specifically, as an optional implementation manner, please refer to Figure 1 , the power supply module includes a power adapter and a power output module;

[0138] The power adapter is used to convert the commercial power into a first-rated DC power supply;

[0139] The power output module is used to convert the first-rated DC power supply into a second-rated DC power supply, a third-rated DC power supply, and a fourth-rated DC power supply;

[0140] Among them, the second-rated DC power supply and the third-rated DC power supply supply power to the first-stage amplification circuit; the second-rated DC power supply and the third-rated DC power supply supply power to the second-stage amplification circuit; the fourth-rated DC power supply supplies power to the bipolar pulse electroporation module.

[0141] In the embodiment of the present application, as Figure 1 shown, the power supply module mainly includes a power adapter and a power output module. Among them, the power adapter is mainly used to convert the input commercial power into a stable first-rated DC power supply. Multiple voltage conversion circuits are provided in the power output module to convert the first-rated DC power supply into a second-rated DC power supply, a third-rated DC power supply, and a fourth-rated DC power supply.

[0142] Exemplarily, please refer to Figure 5 , Figure 5It is a schematic structural diagram of a power module provided by an embodiment of the present application. After the commercial power is input to the power adapter, it will be converted into a DC power supply with a first rated value, and further connect the DC power supply with the first rated value to the power output module. Among them, the power output module can be provided with a TPS5430 converter chip and form a buck circuit with multiple peripheral devices to convert the DC power supply with the first rated value into a DC power supply with a second rated value and a DC power supply with a third rated value. It can also be provided with a TPS61175 converter chip and form a boost circuit with multiple peripheral devices to convert the DC power supply with the first rated value into a DC power supply with a fourth rated value.

[0143] Furthermore, the DC power supply with the second rated value and the DC power supply with the third rated value can supply power to the first operational amplifier in the first-stage amplifier circuit; the DC power supply with the second rated value and the DC power supply with the third rated value can supply power to the second operational amplifier and the third operational amplifier in the second-stage amplifier circuit; the DC power supply with the fourth rated value is for the fourth operational amplifier, the fifth operational amplifier, and the sixth operational amplifier in the biphasic pulse electroporation module.

[0144] Specifically, as an optional implementation manner, the DC power supply with the first rated value is a +12V DC power supply; the DC power supply with the second rated value is a +5V DC power supply; the DC power supply with the third rated value is a -5V DC power supply; the DC power supply with the fourth rated value is a +18V DC power supply.

[0145] In the embodiment of the present application, the power adapter can convert the commercial power into a +12V DC power supply. The TPS5430 converter chip in the power output module can step down the +12V DC power supply to a +5V DC power supply and a -5V DC power supply, and the PS61175 converter chip in the power output module can step up the +12V DC power supply to a +18V DC power supply, so as to supply power to each level of circuits and modules in the myocardial cell intracellular action potential recording system based on biphasic pulse electroporation.

[0146] As Figure 6 shown, Figure 6 It is a schematic flowchart of a method for recording myocardial cell intracellular action potential based on biphasic pulse electroporation provided by an embodiment of the present application. The method includes the following steps:

[0147] Send a pulse signal; among them, the pulse signal will be received by the biphasic pulse electroporation module and converted into a biphasic pulse signal, and then the biphasic pulse signal will be sent to the myocardial cells through the myocardial cell sensor;

[0148] Receive the intracellular electrical signal of the cardiomyocyte to complete the recording of the intracellular action potential of the cardiomyocyte; wherein, the intracellular electrical signal is collected by the cardiomyocyte sensor and transmitted through a first-stage amplifier circuit, a second-stage amplifier circuit, and a data acquisition card module connected in sequence.

[0149] In the embodiment of the present application, the execution subject of the method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation can be a microcontroller in the system for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation, and the microcontroller can select the stm32 microcontroller.

[0150] Further, the microcontroller is connected to the input end of the biphasic pulse electroporation module, and can send pulse signals to the biphasic pulse electroporation module at a fixed time according to specific usage requirements, so that the biphasic pulse electroporation module converts the pulse signals into biphasic pulse signals, and then sends the biphasic pulse signals to the cardiomyocytes through the cardiomyocyte sensor.

[0151] Therefore, when the cardiomyocyte is stimulated by the biphasic pulse signal, a localized electric field is formed on the cell membrane of the cardiomyocyte to generate nanoscale pores on the cell membrane, and the cardiomyocyte sensor can collect the intracellular electrical signal of the action potential change of the cardiomyocyte after being stimulated.

[0152] Further, after the cardiomyocyte sensor collects the intracellular electrical signal of the cardiomyocyte, the present application takes into account the problem that the signal is vulnerable to noise interference during transmission, filters and amplifies the intracellular electrical signal through a first-stage amplifier circuit and a second-stage amplifier circuit, and transmits it to the data acquisition card module. The analog-to-digital converter of the data acquisition card module converts the intracellular electrical signal into a digital signal and transmits it to the microcontroller. The microcontroller receives the intracellular electrical signal of the cardiomyocyte to complete the recording of the intracellular action potential of the cardiomyocyte. Among them, the microcontroller can display the waveform of the intracellular electrical signal of the cardiomyocyte, and perform data analysis on the intracellular electrical signal, calculate the amplitude of the intracellular electrical signal, so as to record and store the amplitude of the intracellular action potential.

[0153] In practical applications, cardiomyocytes can be cultured through the following steps: select suckling mice, disinfect them with 70% alcohol, and then remove the heart tissue; prepare a nutrient solution, which may be composed of 5ml fetal bovine serum (FBS), 0.5ml double antibody solution and 44.5ml DMEM high-glucose culture medium, mix them thoroughly and pour them into a culture dish to wash the extracted cardiomyocytes to remove impurities and residual blood cells on the cell surface; transfer the washed cardiomyocytes to a vial containing D-Hanks solution and use surgical scissors to cut the tissue into pieces; then, incubate the cut cardiomyocytes with 1ml enzyme mixture (0.07% trypsin and 0.05% type II collagenase), repeat the digestion process 12 times, incubate in a cell culture incubator for 8 minutes each time, and gently shake once every 4 minutes to promote cell dispersion, and remove the supernatant at the same time; after the digestion process is completed, aspirate the cells in the suspension and remove the trypsin to obtain pure cardiomyocytes.

[0154] Furthermore, after obtaining the cardiomyocytes, they need to be cultured in a microelectrode array device. The microelectrode array device can be made on a 20mm×20mm×1mm glass substrate. The electrode layer (10nm Ti / 100nm Au) is deposited by spin coating photoresist, UV exposure, development, magnetron sputtering, and SU-8 spin coating to make an insulating layer. The device is baked and finally assembled. The microelectrode array device is mounted on a PCB using PDMS. Conductive silver glue is connected to the electrode pads. A glass ring serves as a cell culture chamber. The reference electrode is fixed to the plastic cover and connected to the PCB. The pin header is soldered to the PCB. Finally, a biphasic pulse signal is used to stimulate the cardiomyocytes. The collected intracellular electrical signals are transmitted to a microcontroller through a primary amplification circuit, a secondary amplification circuit, and a data acquisition card module. The waveform of the intracellular electrical signals can be displayed on an oscilloscope. The intracellular action potential recorded by the microelectrode array shows a typical depolarization and repolarization process. The amplitude of the intracellular electrical signal can be calculated through further data analysis to determine that the amplitude of the intracellular action potential is between 40-80μV.

[0155] Because this application stimulates cardiomyocytes via a biphasic pulse signal, by introducing two voltage pulses of opposite polarity into the stimulation pulse, it can effectively reduce damage to the cell membrane, maintaining cell integrity and physiological activity. Furthermore, it can more clearly distinguish between the cell's spontaneous electrical activity and stimulation-induced electrical activity, improving the signal-to-noise ratio of intracellular electrical signals and providing richer and more stable information for cellular electrophysiological research, with important application value.

[0156] See also Figure 7 , Figure 7 : This is a hardware structure diagram of an electronic device provided in an embodiment of the present application. The electronic device includes:

[0157] The processor 701 can be implemented in the form of a general-purpose CPU (Central Processing Unit), a microprocessor, an application-specific integrated circuit (ASIC), or one or more integrated circuits, etc., and is used to execute relevant programs to implement the technical solutions provided in the embodiments of the present application;

[0158] The memory 702 can be implemented in the form of a read-only memory (ROM), a static storage device, a dynamic storage device, or a random access memory (RAM), etc. The memory 702 can store an operating system and other application programs. When implementing the technical solutions provided in the embodiments of this specification through software or firmware, the relevant program codes are stored in the memory 702 and are called by the processor 701 to execute the method for recording intracellular action potentials of cardiomyocytes based on biphasic pulse electroporation in the embodiments of the present application;

[0159] The input / output interface 703 is used to implement information input and output;

[0160] The communication interface 704 is used to implement communication interaction between this device and other devices, and can implement communication through wired means (such as USB, network cable, etc.) or wireless means (such as mobile network, WIFI, Bluetooth, etc.);

[0161] The bus 705 transmits information between various components of the device (such as the processor 701, the memory 702, the input / output interface 703, and the communication interface 704);

[0162] Among them, the processor 701, the memory 702, the input / output interface 703, and the communication interface 704 are communicatively connected to each other inside the device through the bus 705.

[0163] The embodiments of the present application also provide a computer-readable storage medium. The computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, it implements the above-mentioned method for recording intracellular action potentials of cardiomyocytes based on biphasic pulse electroporation.

[0164] It can be understood that the content in the above method embodiments is applicable to the embodiments of this storage medium. The functions specifically implemented by the embodiments of this storage medium are the same as those of the above method embodiments, and the beneficial effects achieved are also the same as those of the above method embodiments.

[0165] As a non-transitory computer-readable storage medium, the memory can be used to store non-transitory software programs and non-transitory computer-executable programs. In addition, the memory may include high-speed random access memory, and may also include non-transitory memory, such as at least one magnetic disk storage device, a flash memory device, or other non-transitory solid-state storage devices. In some embodiments, the memory may optionally include a memory remotely disposed relative to the processor, and these remote memories can be connected to the processor through a network. Examples of the above networks include, but are not limited to, the Internet, an intranet, a local area network, a mobile communication network, and combinations thereof.

[0166] An intracellular action potential recording system of cardiomyocytes based on biphasic pulse electroporation provided by an embodiment of the present application can effectively reduce damage to the cell membrane, maintain the integrity and physiological activity of the cells, and better record the changes in electrical signals of the cells before and after stimulation by using a biphasic pulse signal as the electroporation stimulation signal, so as to better record the intracellular action potential and reflect the electrophysiological characteristics of the cells.

[0167] The embodiments described in the embodiments of the present application are for more clearly illustrating the technical solutions of the embodiments of the present application, and do not constitute a limitation on the technical solutions provided by the embodiments of the present application. Those skilled in the art will know that with the evolution of technology and the emergence of new application scenarios, the technical solutions provided by the embodiments of the present application are equally applicable to similar technical problems.

[0168] Those skilled in the art can understand that the technical solutions shown in the figures do not constitute a limitation on the embodiments of the present application, and may include more or fewer steps than shown in the figures, or combine certain steps, or different steps.

[0169] The device embodiments described above are merely illustrative. The units described as separate components may or may not be physically separated, that is, they may be located in one place, or may be distributed to multiple network units. Some or all of the modules can be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0170] Those of ordinary skill in the art can understand that all or some of the steps in the methods disclosed above, and the functional modules / units in the systems and devices can be implemented as software, firmware, hardware, and appropriate combinations thereof.

[0171] In the description of the present application and the above-mentioned drawings, the terms "first", "second", "third", "fourth", etc. (if any) are used to distinguish similar objects and do not necessarily describe a specific order or sequence. It should be understood that the data used in this way can be interchanged under appropriate circumstances so that the embodiments of the present application described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or device that comprises a series of steps or units does not necessarily have to be limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products, or devices.

[0172] It should be understood that in the present application, "at least one (item)" means one or more, and "a plurality" means two or more. "And / or" is used to describe the association relationship of associated objects and indicates that three relationships can exist. For example, "A and / or B" can mean: only A exists, only B exists, and both A and B exist at the same time. Among them, A and B can be singular or plural. The character " / " generally means that the associated objects before and after are in an "or" relationship. "At least one (one) of the following" or its similar expression refers to any combination of these items, including any combination of single item (one) or plural items (ones). For example, at least one (one) of a, b, or c can mean: a, b, c, "a and b", "a and c", "b and c", or "a and b and c", where a, b, c can be single or multiple.

[0173] In several embodiments provided by the present application, it should be understood that the disclosed devices and methods can be implemented in other ways. For example, the device embodiments described above are merely illustrative. For example, the above-mentioned division of units is only a logical function division. In actual implementation, there may be other division methods. For example, multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the displayed or discussed coupling or direct coupling or communication connection to each other can be through some interfaces. The indirect coupling or communication connection of devices or units can be in electrical, mechanical, or other forms.

[0174] The units described above as separate components may or may not be physically separated. The components displayed as units may or may not be physical units, that is, they can be located in one place or distributed to multiple network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0175] In addition, in each embodiment of the present application, the functional units can be integrated into one processing unit, or each unit can exist physically alone, or two or more units can be integrated into one unit. The above-mentioned integrated unit can be implemented in the form of hardware or in the form of a software functional unit.

[0176] If the integrated unit is implemented in the form of a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of the present application, in essence, or the part that contributes to the prior art, or all or part of the technical solution, can be embodied in the form of a software product. The computer software product is stored in a storage medium and includes multiple instructions for causing a computer device (which can be a personal computer, a server, or a network device, etc.) to execute all or part of the steps of the methods in the various embodiments of the present application. The foregoing storage medium includes: various media that can store programs, such as USB flash drives, mobile hard disks, read-only memory (ROM), random access memory (RAM), magnetic disks, or optical discs.

[0177] The preferred embodiments of the embodiments of the present application have been described above with reference to the accompanying drawings, and thus do not limit the scope of the rights of the embodiments of the present application. Any modifications, equivalent replacements, and improvements made by those skilled in the art without departing from the scope and essence of the embodiments of the present application shall be within the scope of the rights of the embodiments of the present application.

Claims

1. An intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation, characterized in that The system includes: a biphasic pulse electroporation module, a cardiomyocyte sensor, a microcontroller, a first-stage amplifier circuit, a second-stage amplifier circuit, a data acquisition card module, and a power supply module; The input end of the biphasic pulse electroporation module is connected to the microcontroller, and the output end of the biphasic pulse electroporation module is connected to the cardiomyocyte sensor. The biphasic pulse electroporation module is configured to receive the pulse signal sent by the microcontroller and convert the pulse signal into a biphasic pulse signal, so as to deliver the biphasic pulse signal to cardiomyocytes through the cardiomyocyte sensor; The cardiomyocyte sensor is used to collect the intracellular electrical signal of cardiomyocytes and transmit the intracellular electrical signal to the input end of the data acquisition card module through the sequentially connected first-stage amplifier circuit and the second-stage amplifier circuit; The output end of the data acquisition card module is connected to the microcontroller. The data acquisition card module is configured to output the received intracellular electrical signal to the microcontroller through an analog-to-digital converter, thereby completing the recording of the intracellular action potential of cardiomyocytes; The power supply module is used to supply power to the first-stage amplifier circuit, the second-stage amplifier circuit, and the biphasic pulse electroporation module.

2. The intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation according to claim 1, wherein The first-stage amplifier circuit includes: a first resistor, a second resistor, a third resistor, a fourth resistor, a first capacitor, a second capacitor, and a first operational amplifier; One end of the first capacitor is connected to the input end of the first-stage amplifier circuit; One end of the first resistor and one end of the second resistor are connected to the other end of the first capacitor; The other end of the first resistor is grounded; the other end of the second resistor is connected to the non-inverting input end of the first operational amplifier; One end of the third resistor is connected to the inverting input end of the first operational amplifier; the other end of the third resistor is connected to the output end of the first operational amplifier; One end of the fourth resistor is connected to the inverting input end of the first operational amplifier; the other end of the fourth resistor is grounded; One end of the second capacitor is connected to the inverting input end of the first operational amplifier; the other end of the second capacitor is connected to the output end of the first operational amplifier; The output end of the first operational amplifier is connected to the output end of the first-stage amplifier circuit.

3. The intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation according to claim 1, wherein The second-stage amplifier circuit includes a fifth resistor, a sixth resistor, a seventh resistor, an eighth resistor, a ninth resistor, a tenth resistor, a third capacitor, a fourth capacitor, a fifth capacitor, a sixth capacitor, a second operational amplifier, and a third operational amplifier; One end of the third capacitor is connected to the input end of the second-stage amplifier circuit; One end of the fifth resistor and one end of the sixth resistor are connected to the other end of the third capacitor; The other end of the fifth resistor is grounded; The other end of the sixth resistor and one end of the seventh resistor are connected to one end of the fourth capacitor; The other end of the fourth capacitor is connected to the output end of the second operational amplifier; The other end of the seventh resistor and one end of the fifth capacitor are connected to the non-inverting input end of the second operational amplifier; The other end of the fifth capacitor is grounded; the inverting input terminal of the second operational amplifier is connected to the output terminal of the second operational amplifier; One end of the eighth resistor is connected to the output terminal of the second operational amplifier; the other end of the eighth resistor is connected to the non-inverting input terminal of the third operational amplifier; One end of the ninth resistor is connected to the inverting input terminal of the third operational amplifier; the other end of the ninth resistor is connected to the output terminal of the third operational amplifier; One end of the tenth resistor is connected to the inverting input terminal of the third operational amplifier; the other end of the tenth resistor is grounded; One end of the sixth capacitor is connected to the inverting input terminal of the third operational amplifier; the other end of the sixth capacitor is connected to the output terminal of the third operational amplifier; The output terminal of the third operational amplifier is connected to the output terminal of the second-stage amplification circuit.

4. The intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation according to claim 1, wherein The microcontroller is an STM32 microcontroller.

5. The intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation according to claim 4, wherein The biphasic pulse electroporation module includes a fourth operational amplifier, a fifth operational amplifier, a sixth operational amplifier, an analog switch, and a multiplexer; The DAC pin of the STM32 microcontroller is connected to the input terminal of the fourth operational amplifier, for converting the pulse signal sent by the DAC pin into a positive-phase pulse signal; The output terminal of the fourth operational amplifier is connected to the analog switch, for transmitting the positive-phase pulse signal to the analog switch; The output terminal of the fourth operational amplifier is also connected to the input terminal of the fifth operational amplifier, for converting the positive-phase pulse signal into a negative-phase pulse signal through the fifth operational amplifier; The output terminal of the fifth operational amplifier is connected to the analog switch, for transmitting the negative-phase pulse signal to the analog switch; The PA5 pin and PA8 pin of the STM32 microcontroller are connected to the analog switch, for regulating the pulse widths of the positive-phase pulse signal and the negative-phase pulse signal to generate a biphasic pulse signal; The output terminal of the analog switch passes through the sixth operational amplifier and is connected to the multiplexer; The IO port pin of the STM32 microcontroller is connected to the multiplexer, for enabling control of the biphasic pulse electroporation module to send the biphasic pulse signal to cardiomyocytes through the cardiomyocyte sensor.

6. The intracellular action potential recording system of cardiomyocytes based on biphasic pulse electroporation according to claim 1, wherein The power supply module includes a power adapter and a power output module; The power adapter is used to convert the commercial power into a DC power supply with a first rated value; The power output module is used to convert the DC power supply with the first rated value into a DC power supply with a second rated value, a DC power supply with a third rated value, and a DC power supply with a fourth rated value; Wherein, the DC power supply with the second rated value and the DC power supply with the third rated value supply power to the first-stage amplification circuit; the DC power supply with the second rated value and the DC power supply with the third rated value supply power to the second-stage amplification circuit; the DC power supply with the fourth rated value supplies power to the biphasic pulse electroporation module.

7. The intracellular action potential recording system for cardiomyocytes based on biphasic pulse electroporation according to claim 6, characterized in that The first rated DC power supply is a +12V DC power supply; the second rated DC power supply is a +5V DC power supply; the third rated DC power supply is a -5V DC power supply; the fourth rated DC power supply is a +18V DC power supply.

8. A method for recording intracellular action potential of cardiomyocytes based on biphasic pulse electroporation, characterized in that, The method includes the following steps: Sending a pulse signal; wherein, the pulse signal is received by the biphasic pulse electroporation module and converted into a biphasic pulse signal, so as to issue the biphasic pulse signal to the cardiomyocytes through the cardiomyocyte sensor; Receiving the intracellular electrical signal of the cardiomyocytes to complete the recording of the intracellular action potential of the cardiomyocytes; wherein, the intracellular electrical signal is collected by the cardiomyocyte sensor and transmitted through a first-stage amplification circuit, a second-stage amplification circuit, and a data acquisition card module that are connected in sequence.

9. An electronic device, characterized in that, The electronic device includes a memory and a processor, the memory stores a computer program, and when the processor executes the computer program, it implements the method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation as claimed in claim 8.

10. A computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by the processor, it implements the method for recording the intracellular action potential of cardiomyocytes based on biphasic pulse electroporation as claimed in claim 8.

Images