Pulse electrotransfection instrument with impedance detection function

Abstract

The utility model discloses a pulse electrotransfection instrument with an impedance detection function. The pulse electrotransfection instrument comprises a pulse generator, a transfection electrode cup and an oscilloscope, the pulse generator provided by the utility model generates a high-voltage pulse transmitted to the load resistor and the transfection electrode cup, the high-voltage pulse is utilized to carry out electrotransfection on a cell solution in the electrode cup, and meanwhile, the oscilloscope is utilized to carry out impedance measurement on the cell solution; after transfection and measurement are completed, transfection parameter classification can be carried out on cells with different impedances according to transfection effects and measurement results. According to the utility model, bipolar pulses can be output during electrotransfection, so that the survival rate of transfected cells is improved, meanwhile, the impedance measurement function can be completed, transfection parameter classification of cells with different impedances is realized, and the optimal transfection parameters of one type of cells can be summarized through a small amount of transfection experiments.

Description

Technical Field

[0001] The utility model relates to the technical field of cell electrophysiology, and particularly to a pulsed electroporator with impedance detection function. Background Art

[0002] Electroporation, also known as electrotransfection, is a powerful and efficient technique for introducing nucleic acids, proteins, and other molecules into various cells by transiently increasing the permeability of cell membranes through the action of high-intensity electric fields, enabling the absorption of exogenous molecules in the surrounding medium.

[0003] The deficiencies in the existing electroporation pulse parameters severely limit the efficiency of electroporation, thus restricting the development of gene therapy. For the special field of high-voltage ultrashort pulse electric fields, the dielectric parameters required for studying the cell electrophysiological response mechanism are still measured under steady-state, low-frequency, or sinusoidal waveforms, which are significantly different from high-voltage transient electric fields and cannot accurately guide the biomedical mechanisms of nuclear perforation and gene electroporation. Moreover, unipolar pulses require a large amount of energy and have a high current peak, causing greater damage to cells. At the same time, there is a wide variety of electroporation pulse parameter combinations. Summarizing the optimal transfection parameters for all different cells requires a large number of transfection experiments, which is extremely time-consuming and laborious. Summary of the Utility Model

[0004] The purpose of the utility model is to provide a pulsed electroporator with impedance detection function, including a pulse generator, a transfection electrode cup, and an oscilloscope.

[0005] The pulse generator is used to generate bipolar voltage pulses.

[0006] The pulse generator is electrically connected to the signal input end of the transfection electrode cup to transmit the bipolar voltage pulses to the transfection electrode cup.

[0007] The transfection electrode cup is used to hold the cell solution to be transfected.

[0008] The oscilloscope is electrically connected to the signal output end of the transfection electrode cup to receive the impedance signal from the transfection electrode cup.

[0009] The pulsed electroporator measures the impedance of the cell solution to obtain the optimal transfection parameters for cells with different impedances.

[0010] Furthermore, the circuit topology of the pulsed electroporator is as follows:

[0011] Denote the end where the positive pole of the DC power supply DC1 is located as end A, the end where the negative pole is located as end B, and end B is grounded.

[0012] Denote the end where the positive pole of the DC power supply DC2 is located as end C, the end where the negative pole is located as end D, and end D is grounded.

[0013] Terminal A is connected to the anode of diode D1, and the cathode of diode D1 is connected to energy storage capacitor C1 and then to terminal B.

[0014] The cathode of diode D1 is connected to the drain of main switch S1, the source of main switch S1 is connected to the drain of tail-cut switch S2, and the source of tail-cut switch S2 is connected to terminal B.

[0015] The cathode of diode D1 is connected to the anode of diode D2, and the cathode of diode D2 is connected to energy storage capacitor C2 and then to the source of main switch S1.

[0016] The cathode of diode D2 is connected to the drain of main switch S3, the source of main switch S3 is connected to the drain of tail-cut switch S4, and the source of tail-cut switch S4 is connected to the source of main switch S1.

[0017] Terminal C is connected to the anode of diode D3, and the cathode of diode D3 is connected to energy storage capacitor C3 and then to terminal D.

[0018] The cathode of diode D3 is connected to the drain of main switch S5, the source of main switch S5 is connected to the drain of tail-cut switch S6, and the source of tail-cut switch S6 is connected to terminal D.

[0019] The cathode of diode D3 is connected to the anode of diode D4, and the cathode of diode D4 is connected to energy storage capacitor C4 and then to the source of main switch S5.

[0020] The cathode of diode D4 is connected to the drain of main switch S7, the source of main switch S7 is connected to the drain of tail-cut switch S8, and the source of tail-cut switch S8 is connected to the source of main switch S5.

[0021] The source of main switch S3 is connected to load R L and then connected to the source of main switch S7.

[0022] The source of main switch S3 is sequentially connected to the transfection electrode cup and the current probe of the oscilloscope and then connected to the source of main switch S7.

[0023] The source of main switch S3 is connected to the voltage probe of the oscilloscope and then connected to the source of main switch S7.

[0024] Furthermore, the main switches S1, S3, S5, S7, the tail-cut switches S2, S4, S6, S8 all adopt MOSFET switches, and their gates are floating.

[0025] Furthermore, the operating states of the pulse generator include a parallel charging mode, a series discharging mode, and a tail-cut switch operating mode.

[0026] Further, when the pulse generator operates in the parallel charging mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The DC power supply DC1 charges the energy storage capacitors C1 and C2 through the diodes D1 and D2. The DC power supply DC2 charges the energy storage capacitors C3 and C4 through the diodes D3 and D4.

[0027] Further, the voltages on the energy storage capacitors C1 and C2 are equal to the output voltage of the DC power supply DC1.

[0028] The voltages on the energy storage capacitors C3 and C4 are equal to the output voltage of the DC power supply DC2.

[0029] Further, when the pulse generator operates in the series discharge mode, the main switches S1, S3, S5, and S7 are turned on, and the tail-cut switches S2, S4, S6, and S8 are turned off. The energy storage capacitors C1, C2, C3, and C4 discharge to the load.

[0030] Further, when the pulse generator operates in the tail-cut switch operation mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The tail-cut switches provide a path for the stray capacitance to discharge.

[0031] The technical effects of the present utility model are beyond doubt. The pulse generator provided by the present utility model generates high-voltage pulses that are transmitted to the load resistor and the transfection electrode cup. These high-voltage pulses are used to electrotransfect the cell solution in the electrode cup. At the same time, the impedance of the cell solution is measured by an oscilloscope. After transfection and measurement are completed, the transfection parameters can be classified for cells with different impedances based on the transfection effect and the measurement results.

[0032] The present utility model can output bipolar pulses during electrotransfection, which improves the survival rate of transfected cells. At the same time, it can complete the function of impedance measurement, realize the classification of transfection parameters for cells with different impedances, and summarize the optimal transfection parameters for a type of cells through a small number of transfection experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 is a schematic structural diagram of the present utility model;

[0034] Figure 2 is a topological structure diagram of a two-stage bipolar Marx pulse generator with tail-cut switches;

[0035] Figure 3It is the equivalent circuit diagram of the charging process of the Marx pulse generator with a two-stage bipolar switch with a tail cut;

[0036] Figure 4 It is the equivalent circuit diagram of the discharging process of the Marx pulse generator with a two-stage bipolar switch with a tail cut;

[0037] Figure 5 It is the equivalent circuit diagram of the working process of the tail cut switch of the Marx pulse generator with a two-stage bipolar switch with a tail cut;

[0038] Figure 6 It is the timing diagram of the control signal and the output pulse waveform of the present invention. Specific embodiments

[0039] The present invention will be further described below in conjunction with embodiments, but it should not be understood that the above-mentioned subject matter scope of the present invention is limited to the following embodiments. Without departing from the above technical idea of the present invention, various substitutions and changes made according to ordinary technical knowledge and customary means in the art shall be included within the protection scope of the present invention.

[0040] Embodiment 1:

[0041] See Figures 1 to 6 , a pulse electroporator with an impedance detection function, comprising a pulse generator, a transfection electrode cup, and an oscilloscope.

[0042] The pulse generator is used to generate bipolar voltage pulses.

[0043] The pulse generator is electrically connected to the signal input end of the transfection electrode cup to transmit bipolar voltage pulses to the transfection electrode cup.

[0044] The transfection electrode cup is used to load the cell solution to be transfected.

[0045] The oscilloscope is electrically connected to the signal output end of the transfection electrode cup to receive the impedance signal from the transfection electrode cup.

[0046] The pulse electroporator measures the impedance of the cell solution to obtain the optimal transfection parameters for cells with different impedances.

[0047] Embodiment 2:

[0048] A pulse electroporator with an impedance detection function, the main technical content is shown in Embodiment 1. Further, the circuit topology of the pulse electroporator is as follows:

[0049] Denote the end where the positive pole of the DC power supply DC1 is located as end A, and the end where the negative pole is located as end B, and end B is grounded.

[0050] Denote the end where the positive pole of the DC power supply DC2 is located as the C end, and the end where the negative pole is located as the D end, and the D end is grounded.

[0051] The A end is connected to the anode of the diode D1, and the cathode of the diode D1 is connected to the energy storage capacitor C1 and then connected to the B end.

[0052] The cathode of the diode D1 is connected to the drain of the main switch S1, the source of the main switch S1 is connected to the drain of the tail cut switch S2, and the source of the tail cut switch S2 is connected to the B end.

[0053] The cathode of the diode D1 is connected to the anode of the diode D2, and the cathode of the diode D2 is connected to the energy storage capacitor C2 and then connected to the source of the main switch S1.

[0054] The cathode of the diode D2 is connected to the drain of the main switch S3, the source of the main switch S3 is connected to the drain of the tail cut switch S4, and the source of the tail cut switch S4 is connected to the source of the main switch S1.

[0055] The C end is connected to the anode of the diode D3, and the cathode of the diode D3 is connected to the energy storage capacitor C3 and then connected to the D end.

[0056] The cathode of the diode D3 is connected to the drain of the main switch S5, the source of the main switch S5 is connected to the drain of the tail cut switch S6, and the source of the tail cut switch S6 is connected to the D end.

[0057] The cathode of the diode D3 is connected to the anode of the diode D4, and the cathode of the diode D4 is connected to the energy storage capacitor C4 and then connected to the source of the main switch S5.

[0058] The cathode of the diode D4 is connected to the drain of the main switch S7, the source of the main switch S7 is connected to the drain of the tail cut switch S8, and the source of the tail cut switch S8 is connected to the source of the main switch S5.

[0059] The source of the main switch S3 is connected to the load R L and then connected to the source of the main switch S7.

[0060] The source of the main switch S3 is successively connected to the transfection electrode cup and the current probe of the oscilloscope and then connected to the source of the main switch S7.

[0061] The source of the main switch S3 is connected to the voltage probe of the oscilloscope and then connected to the source of the main switch S7.

[0062] Example 3:

[0063] A pulse electroporation instrument with impedance detection function, the main technical content is as described in any one of Examples 1 to 2. Further, the main switches S1, S3, S5, S7, the tail cut switches S2, S4, S6, S8 all adopt MOSFET switches, and the gates are floating.

[0064] Example 4:

[0065] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Examples 1 to 3. Further, the working states of the pulse generator include a parallel charging mode, a series discharging mode, and a tail-cut switch working mode.

[0066] Example 5:

[0067] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Examples 1 to 4. Further, when the pulse generator works in the parallel charging mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The DC power supply DC1 charges the energy storage capacitors C1 and C2 through the diodes D1 and D2. The DC power supply DC2 charges the energy storage capacitors C3 and C4 through the diodes D3 and D4.

[0068] Example 6:

[0069] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Examples 1 to 5. Further, the voltage on the energy storage capacitors C1 and C2 is equal to the output voltage of the DC power supply DC1.

[0070] The voltage on the energy storage capacitors C3 and C4 is equal to the output voltage of the DC power supply DC2.

[0071] Example 7:

[0072] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Examples 1 to 6. Further, when the pulse generator works in the series discharging mode, the main switches S1, S3, S5, and S7 are turned on, and the tail-cut switches S2, S4, S6, and S8 are turned off. The energy storage capacitors C1, C2, C3, and C4 discharge to the load.

[0073] Example 8:

[0074] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Examples 1 to 7. Further, when the pulse generator works in the tail-cut switch working mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The tail-cut switches provide a loop for the stray capacitance to discharge.

[0075] Example 9:

[0076] A pulse electroporation instrument with impedance detection function, the main technical content is shown in any one of Embodiments 1 to 8. Further, the method adopted for impedance measurement includes the two-electrode voltammetry method.

[0077] The two-electrode voltammetry method calculates the impedance modulus value by calculating the ratio of the peak-to-peak voltage to the peak-to-peak current at both ends of the cell solution.

[0078] Embodiment 10:

[0079] See Figures 1 to 6 , a pulse electroporation instrument with impedance detection function, includes a pulse generator, a transfection electrode cup, and an oscilloscope.

[0080] The pulse generator is used to generate bipolar voltage pulses.

[0081] The pulse generator is electrically connected to the signal input end of the transfection electrode cup to transmit the bipolar voltage pulse to the transfection electrode cup.

[0082] The transfection electrode cup is used to load the cell solution to be transfected.

[0083] The oscilloscope is electrically connected to the signal output end of the transfection electrode cup to receive the impedance signal from the transfection electrode cup.

[0084] The pulse electroporation instrument measures the impedance of the cell solution to obtain the optimal transfection parameters for cells with different impedances.

[0085] The pulse generator generates high-voltage pulses transmitted to the load resistor and the transfection electrode cup. Using this high-voltage pulse to perform electroporation on the cell solution in the electrode cup, and at the same time measuring the impedance of the cell solution through the oscilloscope. After the transfection and measurement are completed, the transfection parameters of cells with different impedances can be classified according to the transfection effect and measurement results.

[0086] Embodiment 11:

[0087] A pulse electroporation instrument with impedance detection function, the main technical content is shown in Embodiment 10. Further, the circuit topology of the pulse electroporation instrument is as follows:

[0088] Denote the end where the positive pole of the DC power supply DC1 is located as end A, and the end where the negative pole is located as end B, and end B is grounded.

[0089] Denote the end where the positive pole of the DC power supply DC2 is located as end C, and the end where the negative pole is located as end D, and end D is grounded.

[0090] End A is connected to the anode of diode D1, and the cathode of diode D1 is connected to energy storage capacitor C1 and then connected to end B.

[0091] The cathode of diode D1 is connected to the drain of main switch S1. The source of main switch S1 is connected to the drain of tail-cut switch S2. The source of tail-cut switch S2 is connected to terminal B.

[0092] The cathode of diode D1 is connected to the anode of diode D2. The cathode of diode D2 is connected to energy storage capacitor C2 and then connected to the source of main switch S1.

[0093] The cathode of diode D2 is connected to the drain of main switch S3. The source of main switch S3 is connected to the drain of tail-cut switch S4. The source of tail-cut switch S4 is connected to the source of main switch S1.

[0094] Terminal C is connected to the anode of diode D3. The cathode of diode D3 is connected to energy storage capacitor C3 and then connected to terminal D.

[0095] The cathode of diode D3 is connected to the drain of main switch S5. The source of main switch S5 is connected to the drain of tail-cut switch S6. The source of tail-cut switch S6 is connected to terminal D.

[0096] The cathode of diode D3 is connected to the anode of diode D4. The cathode of diode D4 is connected to energy storage capacitor C4 and then connected to the source of main switch S5.

[0097] The cathode of diode D4 is connected to the drain of main switch S7. The source of main switch S7 is connected to the drain of tail-cut switch S8. The source of tail-cut switch S8 is connected to the source of main switch S5.

[0098] The source of main switch S3 is connected to load R L and then connected to the source of main switch S7.

[0099] The source of main switch S3 is successively connected to the transfection electrode cup and the current probe of the oscilloscope and then connected to the source of main switch S7.

[0100] The source of main switch S3 is connected to the voltage probe of the oscilloscope and then connected to the source of main switch S7.

[0101] Example 12:

[0102] A pulsed electroporation instrument with impedance detection function, the main technical content is shown in any one of Examples 10 to 11. Further, the main switches S1, S3, S5, S7, tail-cut switches S2, S4, S6, S8 all adopt MOSFET switches and their gates are floating.

[0103] Example 13:

[0104] A pulse electroporator with impedance detection function, the main technical content can be found in any one of Embodiments 10 to 12. Further, the operating states of the pulse generator include a parallel charging mode, a series discharging mode, and a tail-cut switch operating mode.

[0105] Embodiment 14:

[0106] A pulse electroporator with impedance detection function, the main technical content can be found in any one of Embodiments 10 to 13. Further, when the pulse generator operates in the parallel charging mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The DC power supply DC1 charges the energy storage capacitors C1 and C2 through the diodes D1 and D2. The DC power supply DC2 charges the energy storage capacitors C3 and C4 through the diodes D3 and D4.

[0107] Embodiment 15:

[0108] A pulse electroporator with impedance detection function, the main technical content can be found in any one of Embodiments 10 to 14. Further, the voltage on the energy storage capacitors C1 and C2 is equal to the output voltage of the DC power supply DC1.

[0109] The voltage on the energy storage capacitors C3 and C4 is equal to the output voltage of the DC power supply DC2.

[0110] Embodiment 16:

[0111] A pulse electroporator with impedance detection function, the main technical content can be found in any one of Embodiments 10 to 15. Further, when the pulse generator operates in the series discharging mode, the main switches S1, S3, S5, and S7 are turned on, and the tail-cut switches S2, S4, S6, and S8 are turned off. The diodes are cut off due to reverse bias, and the energy storage capacitors C1, C2, C3, and C4 discharge to the load.

[0112] Embodiment 17:

[0113] A pulse electroporator with impedance detection function, the main technical content can be found in any one of Embodiments 10 to 16. Further, when the pulse generator operates in the tail-cut switch operating mode, the main switches S1, S3, S5, and S7 are turned off, and the tail-cut switches S2, S4, S6, and S8 are turned on. The tail-cut switches provide a loop for the stray capacitance to discharge, which greatly shortens the falling edge of the load output waveform.

[0114] Embodiment 18:

[0115] A pulse electroporator with impedance detection function, the main technical content is shown in any one of Embodiments 10 to 17. Further, the method adopted for impedance measurement includes two-electrode voltammetry.

[0116] The two-electrode voltammetry calculates the impedance modulus value by calculating the ratio of the peak-to-peak voltage to the peak-to-peak current at both ends of the cell solution.

[0117] Embodiment 19:

[0118] See Figures 1 to 6 , a pulse electroporator with impedance detection function, the main technical content includes:

[0119] See Figure 1 , the positive and negative electrodes of the pulse generator are respectively connected to the positive and negative electrodes of the load resistor R L . The electrode cups are connected in parallel to the load resistor. The voltage probe of the oscilloscope is connected in parallel across the two ends of the electrode cup, and the current probe of the oscilloscope is connected in series between the negative electrode of the electrode cup and the negative electrode of the pulse generator.

[0120] See Figure 2 , the upper part of the circuit is the positive-pulse generating circuit, and the lower part of the circuit is the negative-pulse generating circuit, which are respectively connected to the positive and negative electrodes of the load resistor R L The positive / negative pulse generating circuit is composed of two-stage energy storage modules connected in parallel, separated by unidirectionally conducting diodes D1, D2, D3, D4, and a 1 kV power supply is provided by DC high-voltage power supplies DC1 and DC2. The whole circuit can generate bipolar voltage pulses (2 kV) twice that of the DC high-voltage power supply. Semiconductor switches (main switches S1, S3, S5, S7, tail-cut switches S2, S4, S6, S8) are connected to an external signal generator, and corresponding control signals are input to control the switch conduction time, thereby controlling the pulse length output to the load resistor.

[0121] Working principle:

[0122] The pulse generator generates high-voltage pulses that are transmitted to the load resistor and the transfection electrode cup. The high-voltage pulses are used to perform electroporation on the cell solution in the electrode cup. At the same time, the impedance of the cell solution is measured through an oscilloscope. After transfection and measurement are completed, transfection parameters can be classified for cells with different impedances based on the transfection effect and measurement results.

[0123] The instrument can be roughly divided into two parts, one is the pulse generator part, and the other is the impedance measurement part.

[0124] In the pulse generator section, the working principle of the Marx pulse generator with a tail-cut switch can be summarized as: "parallel charging, series discharging". The turn-off of the semiconductor switch is achieved by inputting a control signal to the semiconductor switch using a signal generator. Taking the positive pulse as an example:

[0125] Parallel charging process. The main switches S1 and S3 of each stage are turned off, while the tail-cut switches S2 and S4 of each stage are in the conducting state. The high-voltage DC power supply charges the main capacitors C1 and C2 through the diodes D1 and D2. The capacitor voltage on each stage is equal to the output voltage of the high-voltage DC power supply: V_in = V_C1 = V_C2.

[0126] Series discharging process. When the parallel energy storage capacitors of each stage are charged, the main switches S1 and S3 of each stage are triggered to conduct, and the tail-cut switches S2 and S4 of each stage are turned off. The diodes D1 and D2 are cut off due to reverse bias. The capacitors of each stage and the main switch form a series circuit to discharge the load. At this time, a positive pulse of 2V_in will be formed on the load.

[0127] Working process of the tail-cut switch. When the main switches of each stage in the circuit are turned off, the originally turned-off tail-cut switches of each stage are triggered to conduct. At this time, the tail-cut switch provides a low-impedance loop for the stray capacitance to discharge quickly, which greatly shortens the falling edge of the load output waveform.

[0128] In the impedance measurement section, the working principle of impedance measurement is the two-electrode voltammetry method. The voltage probe is connected in parallel across the two ends of the electrode cup, and the current probe is connected in series between the negative electrode of the electrode cup and the negative electrode of the pulse generator. The impedance modulus value is calculated by measuring the ratio of the peak-to-peak voltage to the peak-to-peak current across the cell solution using an oscilloscope.

Claims

1. A pulse electroporator with impedance detection function, characterized in that, It includes a pulse generator, a transfection electrode cup, and an oscilloscope; The pulse generator is used to generate bipolar voltage pulses; The pulse generator is electrically connected to the signal input end of the transfection electrode cup to transmit the bipolar voltage pulses to the transfection electrode cup; The transfection electrode cup is used to load the cell solution to be transfected; The oscilloscope is electrically connected to the signal output end of the transfection electrode cup to receive the impedance signal from the transfection electrode cup; The pulsed electroporator measures the impedance of the cell solution to obtain the optimal transfection parameters for cells with different impedances.

2. The pulsed electroporation instrument with impedance detection function according to claim 1, wherein, The circuit topology of the pulsed electroporator is as follows: Denote the end where the positive pole of the DC power supply DC1 is located as terminal A, the end where the negative pole is located as terminal B, and terminal B is grounded; Denote the end where the positive pole of the DC power supply DC2 is located as terminal C, the end where the negative pole is located as terminal D, and terminal D is grounded; Terminal A is connected to the anode of diode D1, and the cathode of diode D1 is connected to energy storage capacitor C1 and then connected to terminal B; The cathode of diode D1 is connected to the drain of main switch S1, the source of main switch S1 is connected to the drain of tail cut switch S2, and the source of tail cut switch S2 is connected to terminal B; The cathode of diode D1 is connected to the anode of diode D2, and the cathode of diode D2 is connected to energy storage capacitor C2 and then connected to the source of main switch S1; The cathode of diode D2 is connected to the drain of main switch S3, the source of main switch S3 is connected to the drain of tail cut switch S4, and the source of tail cut switch S4 is connected to the source of main switch S1; Terminal C is connected to the anode of diode D3, and the cathode of diode D3 is connected to energy storage capacitor C3 and then connected to terminal D; The cathode of diode D3 is connected to the drain of main switch S5, the source of main switch S5 is connected to the drain of tail cut switch S6, and the source of tail cut switch S6 is connected to terminal D; The cathode of diode D3 is connected to the anode of diode D4, and the cathode of diode D4 is connected to energy storage capacitor C4 and then connected to the source of main switch S5; The cathode of diode D4 is connected to the drain of main switch S7, the source of main switch S7 is connected to the drain of tail cut switch S8, and the source of tail cut switch S8 is connected to the source of main switch S5; The source of the main switch S3 is connected to the load R L and then connected to the source of the main switch S7; The source of main switch S3 is sequentially connected to the transfection electrode cup and the current probe of the oscilloscope and then connected to the source of main switch S7; The source of main switch S3 is connected to the voltage probe of the oscilloscope and then connected to the source of main switch S7.

3. The pulse electrotransfection instrument with impedance detection function according to claim 2, characterized in that, The main switches S1, S3, S5, S7, and the tail cut switches S2, S4, S6, S8 all adopt MOSFET switches, and the gates are floating.

4. The pulse electrotransfection instrument with impedance detection function according to claim 2, characterized in that, The working states of the pulse generator include a parallel charging mode, a series discharging mode, and a tail cut switch working mode.

5. The pulse electrotransfection instrument with an impedance detection function according to claim 4, wherein, When the pulse generator works in the parallel charging mode, main switches S1, S3, S5, S7 are turned off, and tail cut switches S2, S4, S6, S8 are turned on. The DC power supply DC1 charges energy storage capacitors C1 and C2 through diodes D1 and D2; the DC power supply DC2 charges energy storage capacitors C3 and C4 through diodes D3 and D4.

6. The pulse electrotransfection instrument with impedance detection function according to claim 5, characterized in that, The voltages on the energy storage capacitors C1 and C2 are equal to the output voltage of the DC power supply DC1; The voltages on the energy storage capacitors C3 and C4 are equal to the output voltage of the DC power supply DC2.

7. The pulsed electroporation instrument with impedance detection function according to claim 4, characterized in that, When the pulse generator operates in the series discharge mode, the main switches S1, S3, S5, and S7 are turned on, and the tail cut switches S2, S4, S6, and S8 are turned off. The energy storage capacitors C1, C2, C3, and C4 discharge the load.

8. A pulse electrotransfection instrument with an impedance detection function according to claim 4, characterized in that, When the pulse generator operates in the tail cut switch operation mode, the main switches S1, S3, S5, and S7 are turned off, and the tail cut switches S2, S4, S6, and S8 are turned on. The tail cut switches provide a loop for the stray capacitance to discharge.

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