A high frequency bipolar non- recoverable electroporation system and method

Abstract

The present application belongs to the field of biomedical engineering, and provides a high-frequency bipolar non-recoverable electroporation system and method. By combining pulse frequency, voltage amplitude and pulse width, suitable electric field parameters for perforating target cells are obtained. In the process of perforating target cells, pulse frequency, voltage amplitude and pulse width are monitored in real time. By analyzing the trend chart of the three data in the time subunit and the corresponding historical data trend chart, the fluctuation deviation value in the time subunit is obtained. Based on the fluctuation deviation value, adjustment and optimization are carried out. By real-time detection of pulse frequency, voltage amplitude and pulse width, the influence of cell perforation can be accurately mastered. By regulating the three data, it is ensured that the electric field parameters can always maintain the best state for cell perforation, and the damage to other cell tissues is reduced.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomedical engineering, and in particular relates to a high-frequency bipolar non-recoverable electroporation system and method. BACKGROUND

[0002] The high-frequency bipolar technology is to perform irreversible perforation on the cell membrane in cells through extremely short but strong composite electric field pulses, and unlike traditional other technologies, the high-frequency bipolar technology mainly relies on the way of cell apoptosis to realize the inactivation of cells, and can selectively kill the required inactivated cells, and causes less damage to the surrounding other tissues.

[0003] A Chinese patent application with the publication number CN105055015B discloses a non-recoverable electroporation system, which comprises an upper information management unit and a high-voltage pulse discharge main circuit, the upper information management unit transmits a control signal to the high-voltage pulse discharge main circuit through a high-frequency pulse generation control unit, a time sequence function control unit, a first optoelectronic isolation unit and a power amplifier circuit; and the high-voltage pulse discharge main circuit transmits a feedback signal back to the upper information management unit through a voltage and current detection circuit, a second optoelectronic isolation unit and a signal filtering unit.

[0004] In the prior art, only the inactivation of biological cells is realized, and in the inactivation process, the damage to the surrounding tissues is reduced, but the process of inactivating the cells is not monitored and optimized in real time, the optimal state cannot be ensured during the cell perforation work, and the inactivated cells are not evaluated, and the data of the inactivated cells can provide data support for subsequent experiments or treatment. SUMMARY

[0005] In order to make up for the deficiencies of the prior art and solve at least one technical problem in the background art.

[0006] The technical scheme adopted by the present application to solve the technical problems is: a high-frequency bipolar non-recoverable electroporation system and method, comprising:

[0007] Step one, based on the type of target cells, set the electric field parameters, the electric field parameters include pulse frequency, voltage amplitude and pulse width;

[0008] Step two, based on the pulse frequency, voltage amplitude and pulse width, process the three data to obtain the electric field parameter representative value, and determine whether it meets the electric perforation of the target cells;

[0009] A1, based on the historical data, process the pulse frequency in the historical data to obtain the pulse frequency representative value;

[0010] A2, based on the size of the target cells in the treatment area, process to obtain the voltage amplitude representative value;

[0011] A3, based on the inner and outer cell membranes in the target cells in the treatment area, obtaining a cell membrane conductivity representative value, and taking the cell membrane conductivity representative value as a pulse width representative value;

[0012] A4, based on the pulse frequency representative value, the voltage amplitude representative value and the pulse width representative value, obtaining an electric field parameter representative value through a formula;

[0013] In A4, the formula is: electric field parameter representative value = (pulse frequency representative value + voltage amplitude representative value + pulse width representative value) * preset proportionality coefficient;

[0014] Step three, real-time monitoring of the reaction between the electric field parameter and the tissue, obtaining feedback information, and adjusting the electric field parameter;

[0015] Step four, obtaining cell inactivation data in the treatment area, and evaluating the treatment area.

[0016] The application further illustrates that in A1, the pulse frequency of different voltage amplitudes and pulse widths in the historical data is processed, and the processing process is as follows:

[0017] A101, square difference values of the pulse frequency of different voltage amplitudes in the historical data are calculated to obtain pulse frequency representative values under different pulse voltage amplitudes;

[0018] A102, square difference values of the pulse frequency of different pulse widths in the historical data are calculated to obtain pulse frequency representative values under different pulse widths;

[0019] A103, the pulse frequency representative values under different pulse voltage amplitudes and the pulse frequency representative values under different pulse widths are summed and averaged to obtain an average value, and the average value is taken as the pulse frequency representative value.

[0020] The application further illustrates that in A2, the target cell image in the treatment area is detected and processed to obtain two-dimensional images of the cell nucleus, cytoplasm and cell membrane in the target cell, and based on the three images of the cell nucleus, cytoplasm and cell membrane in the target cell, a target cell size representative value is obtained, and the target cell size representative value is taken as the voltage amplitude representative value, and the specific processing process is as follows:

[0021] A201, the cell nucleus image in the target cell is converted into a gray image, and the edges of the cell nucleus image are outlined to calculate the area of the cell nucleus image;

[0022] A202, converting the cytoplasm image of the target cell into a gray image, and drawing lines on the edges of multiple organelles in the cytoplasm image, calculating the area of each organelle image, summing up the area of each organelle image to obtain the area of the cytoplasm image;

[0023] A203, converting the cell membrane image of the target cell into two gray images, and drawing lines on the edges of the two cell membrane images, calculating the area of each cell membrane image, summing up the area of the two cell membrane images to obtain the area of the cell membrane image;

[0024] A204, based on the sum of the three data of the nuclear image area, the cytoplasm image area and the cell membrane image area, the cell image area = the nuclear image area + the cytoplasm image area + the cell membrane image area, the cell image area is taken as the representative value of the target cell image size.

[0025] The application further illustrates that in A3, the inner and outer cell membranes in the target cell in the treatment area are processed to obtain the ion distribution state value in the cell membrane and the ion distribution state value outside the cell membrane, respectively, based on the ion distribution state value in the cell membrane and the ion distribution state value outside the cell membrane, the ion potential difference value inside and outside the cell is obtained, and the ion potential difference value inside and outside the cell is taken as the representative value of the cell membrane conductivity;

[0026] A301, dividing the area outside the cell membrane into several regions, converting each ion distribution in each region into a two-dimensional scatter plot to obtain the distribution state value of each ion in each region, summing up the distribution state values of multiple ions in each region to obtain the average value, taking the average value as the ion distribution state value of the region, and calculating the squared difference value of each region ion distribution state value, and taking the squared difference value as the ion distribution state value outside the cell membrane;

[0027] A302, dividing the area inside the cell membrane into several regions, converting each ion distribution in each region into a two-dimensional scatter plot to obtain the distribution state value of each ion in each region, summing up the distribution state values of multiple ions in each region to obtain the average value, taking the average value as the ion distribution state value of the region, and calculating the squared difference value of each region ion distribution state value, and taking the squared difference value as the ion distribution state value inside the cell membrane;

[0028] A303, based on the ion distribution state value outside the cell membrane and the ion distribution state value inside the cell membrane, the ion distribution state value outside the cell membrane and the ion distribution state value inside the cell membrane are processed, the absolute value of the difference is obtained, and the absolute value is multiplied by a preset proportion coefficient to obtain the ion potential difference value inside and outside the cell membrane.

[0029] The application further illustrates that in step three, according to real-time monitoring of the pulse frequency representative value, the pulse width representative value and the voltage amplitude representative value, the pulse frequency representative value trend graph, the pulse width representative value trend graph and the voltage amplitude representative value trend graph are obtained respectively, and the specific processing process is as follows:

[0030] B1, the pulse frequency representative value trend graph is compared with the historical pulse frequency representative value trend graph, in the time subunit, the data of the corresponding pulse frequency representative value trend graph and the historical pulse frequency representative value trend graph are subtracted, and the difference value is taken as the pulse frequency fluctuation deviation value;

[0031] B2, the pulse width representative value trend graph is compared with the historical pulse width representative value trend graph, in the time subunit, the data of the corresponding pulse width representative value trend graph and the historical pulse width representative value trend graph are subtracted, and the difference value is taken as the pulse width fluctuation deviation value;

[0032] B3, the voltage amplitude representative value trend graph is compared with the historical voltage amplitude representative value trend graph, in the time subunit, the data of the corresponding voltage amplitude representative value trend graph and the historical voltage amplitude representative value trend graph are subtracted, and the difference value is taken as the voltage amplitude fluctuation deviation value;

[0033] B4, based on the pulse frequency fluctuation deviation value, the pulse width fluctuation deviation value and the voltage amplitude fluctuation deviation value, the adjusted pulse frequency representative value trend graph, the pulse width representative value trend graph and the voltage amplitude representative value trend graph are compared with the corresponding historical value trend graph, and the operation is repeated.

[0034] The application further illustrates that in step four, the treatment area is divided into several areas, the cells in each area are image processed, and the required inactivated cells are marked with color to obtain the number of required inactivated cells, and the specific process is as follows:

[0035] C1, the size of the inactivated cells is compared with the size of the target cells to determine whether the cell morphology changes;

[0036] C2, based on the condition that the cell morphology does not change, the internal and external ion potential difference of the inactivated cells is obtained, and the internal and external ion potential difference of the inactivated cells is compared with the internal and external ion potential difference of the target cells to determine whether the cell membrane is damaged;

[0037] C3, the data of the inactivated cells is recorded, the data of the inactivated cells is compared with the data of the required inactivated cells, and the inactivation effect is evaluated.

[0038] The application further illustrates that in C1, the specific process is as follows:

[0039] C101, based on the calculation method of the target cell area, the area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the inactivated cell are obtained;

[0040] C102, based on the area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the inactivated cell, the three images are subtracted from the corresponding area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the target cell, the difference is summed and averaged, and the average value is taken as the representative value of the inactivated cell size;

[0041] C103, the representative value of the inactivated cell size is compared with the preset judgment cell area change threshold value, and the comparison process is as follows:

[0042] If the representative value of the inactivated cell size is greater than or equal to the preset judgment cell area change threshold value, it is determined that the cell morphology has changed, and it belongs to the inactivated cell;

[0043] If the representative value of the inactivated cell size is less than the preset judgment cell area change threshold value, it is determined that the cell morphology has not changed, and it does not belong to the inactivated cell.

[0044] The application further illustrates that in C2, the specific processing process is as follows:

[0045] C201, based on the calculation method of the ion potential difference inside and outside the target cell, the ion potential difference value inside and outside the inactivated cell is obtained;

[0046] C202, the ion potential difference value inside and outside the inactivated cell is subtracted from the corresponding ion potential difference value inside and outside the target cell, and the difference is taken as the representative value of the cell membrane inside and outside the cell;

[0047] C203, the representative value of the cell membrane inside and outside the cell is compared with the preset judgment cell membrane inside and outside the cell damage threshold value, and the comparison process is as follows:

[0048] If the representative value of the cell membrane inside and outside the cell is greater than or equal to the judgment cell membrane inside and outside the cell damage threshold value, it is determined that the cell membrane inside and outside the cell is damaged, and it belongs to the inactivated cell;

[0049] If the representative value of the cell membrane inside and outside the cell is less than the judgment cell membrane inside and outside the cell damage threshold value, it is determined that the cell membrane inside and outside the cell is not damaged, and it does not belong to the inactivated cell.

[0050] The application further illustrates that in C3, the comparison process is as follows:

[0051] If the data of the inactivated cell is consistent with the required inactivated cell, it is determined that the evaluation data is consistent;

[0052] If the data of the inactivated cells is inconsistent with the data of the desired inactivated cells, it is determined that the evaluation data is not in conformity.

[0053] The application further discloses that the data acquisition module sets the electric field parameters based on the type of the target cells, and the electric field parameters include the pulse frequency, the voltage amplitude and the pulse width.

[0054] The data processing and analysis module processes the three data of the pulse frequency, the voltage amplitude and the pulse width, obtains the representative value of the electric field parameters, and determines whether the electric field parameters meet the requirements of the electroporation of the target cells.

[0055] A1, based on the historical data, the pulse frequency in the historical data is processed to obtain the representative value of the pulse frequency.

[0056] A2, based on the size of the target cells in the treatment area, the voltage amplitude representative value is obtained by processing.

[0057] A3, based on the inner and outer cell membranes in the target cells in the treatment area, the cell membrane conductivity representative value is obtained by processing, and the cell membrane conductivity representative value is taken as the pulse width representative value.

[0058] A4, based on the three data of the pulse frequency representative value, the voltage amplitude representative value and the pulse width representative value, the electric field parameter representative value is obtained by a formula.

[0059] In A4, the formula is: electric field parameter representative value = (pulse frequency representative value + voltage amplitude representative value + pulse width representative value) × preset proportionality coefficient.

[0060] The monitoring and optimization module monitors the reaction between the electric field parameters and the tissues in real time, obtains feedback information, and adjusts the electric field parameters.

[0061] The self-evaluation module obtains the inactivation data of the cells in the treatment area, and evaluates the treatment area.

[0062] The application has the following beneficial effects:

[0063] 1. The high-frequency bipolar non-recoverable electroporation system and method of the present application, by combining the pulse frequency, voltage amplitude and pulse width three data, get the electric field parameters suitable for perforating the target cells, and in the process of perforating the target cells, real-time monitoring the pulse frequency, voltage amplitude and pulse width three data, by analyzing the three data trend chart in the time subunit and the corresponding historical data trend chart, get the fluctuation deviation value in the time subunit, based on the fluctuation deviation value for adjustment and optimization, through real-time detection of pulse frequency, voltage amplitude and pulse width three data, can accurately master the influence of cell perforation, and regulate the three data, ensure that the electric field parameters can keep the best state for cell perforation work all the time, reduce the damage to other cell tissues;

[0064] 2. The high-frequency bipolar non-recoverable electroporation system and method of the present application, by comparing the size of the inactivated cells and the size of the target cells, determining whether the cell morphology changes, based on the condition that the cell morphology does not change, obtaining the internal and external ion potential difference of the inactivated cells, comparing the internal and external ion potential difference of the inactivated cells with the internal and external ion potential difference of the target cells, determining whether the cell membrane is damaged, recording the data of the inactivated cells, comparing the data of the inactivated cells with the data of the required inactivated cells, evaluating the effect of the inactivated cells, through this method, not only can record and analyze a large amount of inactivated cell data to provide data support for subsequent experiments or treatment, but also can develop personalized inactivation strategies for specific types of cells or individuals, improve the pertinence of treatment. BRIEF DESCRIPTION OF DRAWINGS

[0065] The present application will be further described below with reference to the accompanying drawings.

[0066] Figure 1 is the flow chart of the first embodiment of the present application;

[0067] Figure 2 is the flow chart of the second embodiment of the present application;

[0068] Figure 3 is the system module diagram of the present application. DETAILED DESCRIPTION

[0069] In order to make the technical means, creative features, purposes and effects realized by the present application easy to understand, the present application will be further described below with reference to the specific embodiments. Embodiment one

[0070] As shown in Figure 1 , the high-frequency bipolar non-recoverable electroporation method of the embodiment of the present application comprises the following steps:

[0071] Step one, set the electric field parameters based on the type of target cells, the electric field parameters include pulse frequency, voltage amplitude and pulse width;

[0072] Step two, process the three data of pulse frequency, voltage amplitude and pulse width, get the representative value of electric field parameters, and determine whether it meets the electric perforation of target cells;

[0073] In step two, based on the type of target cells in the treatment area, the three data of pulse frequency, voltage amplitude and pulse width are processed to obtain the electric field parameters suitable for electric perforation of target cells;

[0074] A1, based on historical data, process the pulse frequency in the historical data to obtain the representative value of pulse frequency;

[0075] In A1, the pulse frequency of different voltage amplitudes and pulse widths in the historical data is processed, and the processing process is as follows:

[0076] A101, square difference value of pulse frequency of different voltage amplitudes in historical data is calculated to obtain the representative value of pulse frequency under different pulse voltage amplitudes;

[0077] A102, square difference value of pulse frequency of different pulse widths in historical data is calculated to obtain the representative value of pulse frequency under different pulse widths;

[0078] A103, the representative value of pulse frequency under different pulse voltage amplitudes and the representative value of pulse frequency under different pulse widths are summed and averaged, and the average value is taken as the representative value of pulse frequency;

[0079] A2, process based on the size of target cells in the treatment area to obtain the representative value of voltage amplitude;

[0080] In A2, the target cell image in the treatment area is detected and processed to obtain the two-dimensional images of the nucleus, cytoplasm and cell membrane in the target cell, and based on the three images of the nucleus, cytoplasm and cell membrane in the target cell, the representative value of the size of the target cell is obtained, which is taken as the representative value of the voltage amplitude, and the specific processing process is as follows:

[0081] A201, convert the nucleus image in the target cell into a gray image, and draw the edge of the nucleus image to calculate the area of the nucleus image;

[0082] In some specific embodiments, the gray images of the nucleus at different angles in the cell are obtained, the edge of each nucleus image is drawn, and the areas of the multiple nucleus images at different angles are summed and averaged, and the average value is taken as the area of the nucleus image;

[0083] A202, converting the cytoplasm image in the target cell into a gray image, and drawing lines on the edges of multiple organelles in the cytoplasm image, calculating the area of each organelle image, summing up the area of each organelle image to obtain the area of the cytoplasm image;

[0084] In some embodiments, by acquiring images of each organelle at different angles, drawing lines on the different angle images of the organelle and calculating the area, summing up the areas of multiple organelle images at different angles and taking the average value, the average value is taken as the area of the organelle image;

[0085] Based on the area of each cell image, the areas of multiple organelle images in the cytoplasm are summed and averaged, and the average value is taken as the area of the cytoplasm image;

[0086] A203, converting the cell membrane image in the target cell into two gray images, and drawing lines on the edges of the two cell membrane images, calculating the area of each cell membrane image, summing up the areas of the two cell membrane images to obtain the area of the cell membrane image;

[0087] Based on the above calculation of the area of the cell nucleus image, the area of the cytoplasm image and the area of the cell membrane image, in some embodiments, a two-dimensional coordinate system is established, in which the image edge lines are divided into a plurality of line segments, the length of each line segment is calculated, and the length and width values in the two-dimensional image are obtained by summing up the length of each line segment;

[0088] Based on the length and width values in the two-dimensional image, the area of the two-dimensional image is obtained according to the area formula: area=length x width;

[0089] A204, based on the sum of the area of the cell nucleus image, the area of the cytoplasm image and the area of the cell membrane image, the area of the cell image = the area of the cell nucleus image + the area of the cytoplasm image + the area of the cell membrane image, and the area of the cell image is taken as the representative value of the size of the target cell image;

[0090] A3, based on the treatment of the inner and outer cell membranes in the target cells in the treatment area, the representative value of the cell membrane conductivity is obtained, and the representative value of the cell membrane conductivity is taken as the representative value of the pulse width;

[0091] In A3, the inner and outer cell membranes in the target cells in the treatment area are treated to obtain the ion distribution state value inside the cell membrane and the ion distribution state value outside the cell membrane, respectively, and based on the two data of the ion distribution state value inside the cell membrane and the ion distribution state value outside the cell membrane, the ion potential difference value inside and outside the cell is obtained, and the ion potential difference value inside and outside the cell is taken as the representative value of the cell membrane conductivity;

[0092] A301, divide the extracellular membrane region into several regions, convert the distribution of each ion in each region into a two-dimensional scatter plot, obtain the distribution state value of each ion in each region, sum and average the distribution state values of multiple ions in each region, take the average value as the regional ion distribution state value, and calculate the squared difference value of each regional ion distribution state value as the ion distribution state value outside the cell membrane;

[0093] A302, divide the intracellular membrane region into several regions, convert the distribution of each ion in each region into a two-dimensional scatter plot, obtain the distribution state value of each ion in each region, sum and average the distribution state values of multiple ions in each region, take the average value as the regional ion distribution state value, and calculate the squared difference value of each regional ion distribution state value as the ion distribution state value inside the cell membrane;

[0094] A303, based on the ion distribution state value outside the cell membrane and the ion distribution state value inside the cell membrane, the formula is: obtain the ion potential difference E between the inside and outside of the cell membrane, Q represents the ion distribution state value outside the cell membrane, P represents the ion distribution state value inside the cell membrane, and a represents a preset proportion coefficient;

[0095] A4, based on the pulse frequency representative value, the voltage amplitude representative value and the pulse width representative value, the electric field parameter representative value is obtained by the formula;

[0096] In A4, the electric field parameter representative value W is obtained by the formula: W=(L+E+K)×β, L represents the pulse frequency representative value, K represents the voltage amplitude representative value, and β represents a preset proportion coefficient;

[0097] Step three, real-time monitor the reaction between the electric field parameter and the tissue, obtain feedback information, and adjust the electric field parameter;

[0098] In step three, according to the real-time monitoring and processing of the pulse frequency representative value, the pulse width representative value and the voltage amplitude representative value, the pulse frequency representative value trend graph, the pulse width representative value trend graph and the voltage amplitude representative value trend graph are obtained respectively, and the specific processing process is as follows:

[0099] B1, overlap and compare the pulse frequency representative value trend graph with the historical pulse frequency representative value trend graph, in the time subunit, subtract the data of the corresponding pulse frequency representative value trend graph from the historical pulse frequency representative value trend graph, and take the difference value as the pulse frequency fluctuation deviation value;

[0100] B2, Overlay and compare the pulse width representative value change trend chart with the historical pulse width representative value change trend chart. Within the time sub-unit, the difference between the corresponding pulse width representative value change trend chart and the historical pulse width representative value change trend chart is calculated, and the difference is used as the pulse width fluctuation deviation value.

[0101] B3. Overlay and compare the voltage amplitude representative value change trend chart with the historical voltage amplitude representative value change trend chart. Within the time sub-unit, subtract the data of the corresponding voltage amplitude representative value change trend chart from the historical voltage amplitude representative value change trend chart, and use the difference as the voltage amplitude fluctuation deviation value.

[0102] B4. Adjustments are made based on the pulse frequency fluctuation deviation, pulse width fluctuation deviation, and voltage amplitude fluctuation deviation. The adjusted pulse frequency representative value change trend chart, pulse width representative value change trend chart, and voltage amplitude representative value change trend chart are then compared with the corresponding historical value change trend charts, and this process is repeated.

[0103] The technical solution of Embodiment 1 of the present invention is as follows: By combining three data points—pulse frequency, voltage amplitude, and pulse width—suitable electric field parameters for perforating target cells are obtained. During the perforation of target cells, the three data points are monitored in real time. By analyzing the trend graph of the three data points within a time sub-unit and the corresponding historical data trend graph, the fluctuation deviation value within the time sub-unit is obtained. Based on the fluctuation deviation value, adjustments and optimizations are made. By real-time monitoring of the three data points—pulse frequency, voltage amplitude, and pulse width—the impact on cell perforation can be accurately grasped. By regulating the three data points, it is ensured that the electric field parameters can always maintain the optimal state for cell perforation, reducing damage to other cells and tissues. Example 2

[0104] Step 4: Obtain cell inactivation data within the treatment area and evaluate the treatment area;

[0105] like Figure 2 As shown, in step four, the treatment area is divided into several regions, the cells in each region are image-processed, and the cells to be inactivated are marked with color to obtain the required number of inactivated cells. The specific process is as follows:

[0106] C1, the size of inactivated cells is monitored and compared with the size of target cells to determine whether cell morphology has changed. The specific process is as follows:

[0107] C101, based on the method of calculating the area of ​​the target cell, yields the area of ​​the cell nucleus, the area of ​​the cytoplasm, and the area of ​​the cell membrane in the inactivated cell;

[0108] C102, based on the area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the cell after inactivation, subtracting the area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the corresponding target cell from the three images, summing the difference values and taking the average value as the representative value of the cell size after inactivation;

[0109] C103, comparing the representative value of the cell size after inactivation with a preset threshold value for determining the change in cell area, and the comparison process is as follows:

[0110] If the representative value of the cell size after inactivation is greater than or equal to the preset threshold value for determining the change in cell area, it is determined that the cell morphology has changed and belongs to the inactivated cell.

[0111] If the representative value of the cell size after inactivation is less than the preset threshold value for determining the change in cell area, it is determined that the cell morphology has not changed and does not belong to the inactivated cell.

[0112] C2, based on the case where the cell morphology has not changed, obtaining the internal and external ion potential difference of the cell after inactivation, comparing the internal and external ion potential difference of the cell after inactivation with the internal and external ion potential difference of the target cell, and determining whether the internal and external cell membrane of the cell is damaged.

[0113] C201, based on the method of obtaining the internal and external ion potential difference of the target cell, obtaining the internal and external ion potential difference value of the cell after inactivation.

[0114] C202, subtracting the internal and external ion potential difference value of the cell after inactivation from the corresponding internal and external ion potential difference value of the target cell, and taking the difference value as the representative value of the internal and external cell membrane.

[0115] C203, comparing the representative value of the internal and external cell membrane with a preset threshold value for determining the damage of the internal and external cell membrane, and the comparison process is as follows:

[0116] If the representative value of the internal and external cell membrane is greater than or equal to the threshold value for determining the damage of the internal and external cell membrane, it is determined that the internal and external cell membrane is damaged and belongs to the inactivated cell.

[0117] If the representative value of the internal and external cell membrane is less than the threshold value for determining the damage of the internal and external cell membrane, it is determined that the internal and external cell membrane is not damaged and does not belong to the inactivated cell.

[0118] C3, recording the data of the inactivated cell, comparing the data of the inactivated cell with the data of the required inactivated cell, and the specific process is as follows:

[0119] If the data of the inactivated cell is consistent with the data of the required inactivated cell, it is determined that the evaluation data is consistent.

[0120] If the data of the inactivated cell is not consistent with the data of the required inactivated cell, it is determined that the evaluation data is not consistent.

[0121] The second embodiment of the present application technical solution: by monitoring the size of the inactivated cells and the size of the target cells, it is determined whether the cell morphology has changed, based on the case that the cell morphology has not changed, the internal and external ion potential difference of the inactivated cells is obtained, the internal and external ion potential difference of the inactivated cells is compared with the internal and external ion potential difference of the target cells, it is determined whether the cell membrane is damaged, the data of the inactivated cells is recorded, the data of the inactivated cells is compared with the data of the required inactivated cells, and the effect of the inactivated cells is evaluated. Through this method, not only can a large amount of inactivated cell data be recorded and analyzed to provide data support for subsequent experiments or treatment, but also personalized inactivation strategies can be developed for specific types of cells or individuals, improving the targeting of treatment. Embodiment three

[0122] As shown in the figure, a high-frequency bipolar non-recoverable electroporation system includes: Figure 3

[0123] The data acquisition module sets the electric field parameters based on the type of target cells, including pulse frequency, voltage amplitude and pulse width;

[0124] The data processing and analysis module processes the three data of pulse frequency, voltage amplitude and pulse width to obtain the representative value of electric field parameters, and determines whether it meets the requirements of electroporation on target cells;

[0125] A1, based on historical data, the pulse frequency in the historical data is processed to obtain the representative value of pulse frequency;

[0126] A2, based on the size of the target cells in the treatment area, the representative value of voltage amplitude is obtained;

[0127] A3, based on the internal and external cell membranes in the target cells in the treatment area, the representative value of cell membrane conductivity is obtained, and the representative value of cell membrane conductivity is taken as the representative value of pulse width;

[0128] A4, based on the three data of pulse frequency representative value, voltage amplitude representative value and pulse width representative value, the electric field parameter representative value is obtained through the formula;

[0129] In A4, the formula is: electric field parameter representative value = (pulse frequency representative value + voltage amplitude representative value + pulse width representative value) x preset proportionality coefficient;

[0130] The monitoring and optimization module real-time monitors the reaction between the electric field parameters and the tissue, obtains feedback information, and adjusts the electric field parameters.

[0131] The self-evaluation module obtains the inactivation data of cells in the treatment area and evaluates the treatment area The self-evaluation module obtains the inactivation data of cells in the treatment area and evaluates the treatment area

[0132] The foregoing presents and describes the basic principles, main features and advantages of the present application. It should be understood by those skilled in the art that the present application is not limited to the above-mentioned embodiments, and the above-mentioned embodiments and descriptions in the specification are only to illustrate the principles of the present application. Without departing from the spirit and scope of the present application, various changes and improvements can be made to the present application, and these changes and improvements all fall within the scope of the present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. A high frequency bipolar non- recoverable electroporation system, characterized by: Comprising: a data acquisition module, based on the type of target cells, set the electric field parameters, including pulse frequency, voltage amplitude and pulse width; a data processing and analysis module, based on the pulse frequency, voltage amplitude and pulse width three data processing, get the electric field parameter representative value, and determine whether to meet the target cells for electroporation; A1, based on historical data, the pulse frequency in the historical data is processed to obtain the pulse frequency representative value; A2, based on the size of the target cells in the treatment area, the voltage amplitude representative value is obtained by processing; A3, based on the inner and outer cell membranes in the target cells in the treatment area, the cell membrane conductivity representative value is obtained, and the cell membrane conductivity representative value is taken as the pulse width representative value; A4, based on the pulse frequency representative value, the voltage amplitude representative value and the pulse width representative value three data, the electric field parameter representative value is obtained by formula; In A4, the formula is: electric field parameter representative value=(pulse frequency representative value+ voltage amplitude representative value+ pulse width representative value)× preset proportionality coefficient; a monitoring and optimization module, which monitors the reaction between the electric field parameters and the tissue in real time, obtains feedback information, and adjusts the electric field parameters; a self-evaluation module, which obtains cell inactivation data in the treatment area and evaluates the treatment area.

2. The high-frequency bipolar non-recoverable electroporation system according to claim 1, wherein: In A1, the pulse frequency of different voltage amplitudes and pulse widths in the historical data is processed, and the processing process is as follows: A101, square difference value is calculated for the pulse frequency of different voltage amplitudes in the historical data, and the pulse frequency representative value under different pulse voltage amplitudes is obtained; A102, square difference value is calculated for the pulse frequency of different pulse widths in the historical data, and the pulse frequency representative value under different pulse widths is obtained; A103, the pulse frequency representative value under different pulse voltage amplitudes and the pulse frequency representative value under different pulse widths are summed and averaged, and the average value is taken as the pulse frequency representative value.

3. The high-frequency bipolar non-recoverable electroporation system according to claim 1, wherein: In A2, the target cell image in the treatment area is detected and processed to obtain the two-dimensional images of the nucleus, cytoplasm and cell membrane in the target cell, and based on the three images of the nucleus, cytoplasm and cell membrane in the target cell, the target cell size representative value is obtained, which is taken as the voltage amplitude representative value, and the specific processing process is as follows: A201, the nucleus image in the target cell is converted into a gray image, and the edge of the nucleus image is outlined, and the area of the nucleus image is calculated; A202, the cytoplasm image in the target cell is converted into a gray image, and the edges of multiple organelles in the cytoplasm image are outlined, and the area of each organelle image is calculated, and the areas of each organelle image are summed to obtain the area of the cytoplasm image; A203, convert the cell membrane image in the target cell into two gray images, and draw the edges of the two cell membrane images, calculate the area of each cell membrane image, sum the areas of the two cell membrane images to obtain the area of the cell membrane image; A204, based on the sum of the three data of the nuclear image area, the cytoplasm image area and the cell membrane image area, the cell image area = the nuclear image area + the cytoplasm image area + the cell membrane image area, the cell image area is taken as the target cell size representative value.

4. The high-frequency bipolar non-recoverable electroporation system according to claim 1, wherein: In A3, the inner and outer cell membranes in the target cells in the treatment area are processed to obtain the ion distribution state value inside the cell membrane and the ion distribution state value outside the cell membrane, respectively, and based on the ion distribution state value inside the cell membrane and the ion distribution state value outside the cell membrane, the intracellular and extracellular ion potential difference value is obtained, which is taken as the cell membrane conductivity representative value; A301, divide the area outside the cell membrane into several regions, convert each ion distribution in each region into a two-dimensional scatter plot to obtain the distribution state value of each ion in each region, sum and average the distribution state values of multiple ions in each region, and take the average value as the regional ion distribution state value. Square difference value of each regional ion distribution state value, the square difference value is taken as the ion distribution state value outside the cell membrane; A302, divide the area inside the cell membrane into several regions, convert each ion distribution in each region into a two-dimensional scatter plot to obtain the distribution state value of each ion in each region, sum and average the distribution state values of multiple ions in each region, and take the average value as the regional ion distribution state value. Square difference value of each regional ion distribution state value, the square difference value is taken as the ion distribution state value inside the cell membrane; A303, based on the ion distribution state value outside the cell membrane and the ion distribution state value inside the cell membrane, the ion distribution state value outside the cell membrane and the ion distribution state value inside the cell membrane are subtracted to obtain the absolute value, and the absolute value is multiplied by a preset proportion coefficient to obtain the intracellular and extracellular ion potential difference value.

5. The high-frequency bipolar non- recoverable electroporation system according to claim 1, wherein: According to the real-time monitoring and processing of the pulse frequency representative value, the pulse width representative value and the voltage amplitude representative value, the pulse frequency representative value trend graph, the pulse width representative value trend graph and the voltage amplitude representative value trend graph are obtained respectively. The specific processing process is as follows: B1, overlap and compare the pulse frequency representative value trend graph with the historical pulse frequency representative value trend graph, and in the time subunit, subtract the data of the corresponding pulse frequency representative value trend graph from the historical pulse frequency representative value trend graph. The difference value is taken as the pulse frequency fluctuation deviation value; B2, superimposedly comparing the pulse width representative value change trend graph with the historical pulse width representative value change trend graph, and in the time subunit, subtracting the data of the corresponding pulse width representative value change trend graph from the historical pulse width representative value change trend graph, and taking the difference value as the pulse width fluctuation deviation value; B3, superimposedly comparing the voltage amplitude representative value change trend graph with the historical voltage amplitude representative value change trend graph, and in the time subunit, subtracting the data of the corresponding voltage amplitude representative value change trend graph from the historical voltage amplitude representative value change trend graph, and taking the difference value as the voltage amplitude fluctuation deviation value; B4, based on the pulse frequency fluctuation deviation value, the pulse width fluctuation deviation value and the voltage amplitude fluctuation deviation value, adjusting the adjusted pulse frequency representative value change trend graph, the pulse width representative value change trend graph and the voltage amplitude representative value trend graph, and superimposedly comparing the corresponding historical value change trend graph, and repeatedly operating.

6. The high-frequency bipolar non- recoverable electroporation system according to claim 1, wherein: The treatment area is divided into several regions, the cells in each region are image processed, and the required inactivated cells are marked with color to obtain the number of required inactivated cells, and the specific process is as follows: C1, comparing the size of the inactivated cells with the size of the target cells to determine whether the cell morphology has changed; C2, based on the case that the cell morphology has not changed, obtaining the internal and external ion potential difference of the inactivated cells, comparing the internal and external ion potential difference of the inactivated cells with the internal and external ion potential difference of the target cells to determine whether the cell membrane is damaged; C3, recording the data of the inactivated cells, comparing the data of the inactivated cells with the data of the required inactivated cells, and evaluating the inactivation effect.

7. A high-frequency bipolar non- recoverable electroporation system according to claim 6, characterized in that: In C1, the specific process is as follows: C101, based on the method of obtaining the area of the target cell nucleus, the area of the cytoplasm and the area of the cell membrane are obtained; C102, based on the area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the inactivated cells, the three images are subtracted from the corresponding area of the nucleus, the area of the cytoplasm and the area of the cell membrane in the target cells, the difference value is summed and averaged, and the average value is taken as the representative value of the size of the inactivated cells; C103, comparing the representative value of the size of the inactivated cells with the preset judgment cell area change threshold value, and the comparison process is as follows: If the representative value of the size of the inactivated cells is greater than or equal to the preset judgment cell area change threshold value, it is determined that the cell morphology has changed and belongs to the inactivated cells; If the representative value of the size of the inactivated cells is less than the preset judgment cell area change threshold value, it is determined that the cell morphology has not changed and does not belong to the inactivated cells.

8. The high-frequency bipolar non- recoverable electroporation system according to claim 6, characterized in that: In C2, the specific processing process is as follows: C201, based on the method of obtaining the internal and external ion potential difference of the target cell, the internal and external ion potential difference value of the inactivated cell is obtained; C202, subtracting the internal and external ion potential difference value of the inactivated cell from the corresponding internal and external ion potential difference value of the target cell, and taking the difference value as the representative value of the cell membrane; C203, comparing the representative value of the intracellular and extracellular cell membrane with the preset threshold value of judging the damage of the intracellular and extracellular cell membrane, and the comparison process is as follows: If the representative value of the intracellular and extracellular cell membrane is greater than or equal to the threshold value of judging the damage of the intracellular and extracellular cell membrane, it is determined that the intracellular and extracellular cell membrane is damaged, which belongs to the inactivated cell; If the representative value of the intracellular and extracellular cell membrane is less than the threshold value of judging the damage of the intracellular and extracellular cell membrane, it is determined that the intracellular and extracellular cell membrane is not damaged, which does not belong to the inactivated cell.

9. The high-frequency bipolar non- recoverable electroporation system according to claim 6, characterized in that: In C3, the comparison process is as follows: If the data of the inactivated cell is consistent with the required inactivated cell, it is determined that the evaluation data is consistent; If the data of the inactivated cell is not consistent with the required inactivated cell, it is determined that the evaluation data is not consistent.

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