Apparatus and method for measuring electroporation ablation performance using high frequency bidirectional pulses

Abstract

An object of the present invention is to provide an apparatus and method for measuring electroporation ablation performance using high-frequency bidirectional pulses. The apparatus includes a pulse generation mechanism including a bipolar pulse generator and a pulse control unit for generating high-frequency bidirectional pulses to a cell suspension of a target cell cluster; a traversal analysis device for traversing each combination of values ​​of a target pulse frequency, a predetermined electric field strength, and a predetermined duration for a target cell cluster consisting of a certain total number of living cells at a predetermined site, and predicting each corresponding cell inhibition rate and each low-damage ablation performance level; and a best-selection analysis mechanism for selecting the combination of values ​​with the best ablation performance based on the predicted data. This invention introduces an effective and automated design of an electroporation ablation system, and simultaneously uses an artificial intelligence mode and a traversal mode based on numerical simulation to obtain each optimized ablation parameter combination for ablation target structures of different sites and different volumes.

Description

[Technical Field]

[0001] The present invention is in the field of radio frequency bidirectional pulse electroporation, and more particularly relates to an apparatus and method for measuring electroporation ablation performance using radio frequency bidirectional pulses. [Background technology]

[0002] Compared with millimeter-wave bipolar pulses, microwave bipolar pulses have a superior ablation effect when used to ablate various tissues, such as cell clusters in porcine liver. Although microwave bipolar pulses are slightly less effective at increasing the rate of cell necrosis in tissues than millimeter-wave bipolar pulses, they act on mitochondria to disrupt the mitochondrial membrane potential, thereby producing apoptotic proteins such as BAX, BCL-2, caspase-3, and caspase-9.

[0003] In other words, the effect of cell inhibition by millimeter-wave-level high-frequency bipolar pulses is reflected only in the specific value of the cell necrosis rate, whereas the effect of cell inhibition by microwave-level high-frequency bipolar pulses is also reflected in the specific value of the cell necrosis rate, i.e., the form of cell death after the cells in the affected area undergo ablation treatment includes not only necrosis but also apoptosis.

[0004] For example, Patent Document 1 proposes a high-frequency bipolar irreversible electroporation system comprising an upper-level information management module, a lower-level control module, and a bipolar high-voltage pulse discharge circuit. The upper-level information management module receives set operating parameters and transmits them to the lower-level control module, which generates a control signal and transmits it to the bipolar high-voltage pulse discharge circuit to generate a bipolar high-voltage pulse. Compared with conventional unipolar irreversible electroporation pulse sequences, this invention can better and more uniformly raise the induced membrane potential of densely packed cells to the simulated electroporation threshold, thereby achieving a more uniform ablation area. This invention can achieve good tumor ablation effects and tumor growth inhibition goals, and is safe and effective.

[0005] For example, Patent Document 2 proposes a high-frequency irreversible electroporation device. The device includes a power converter circuit, an energy storage circuit, an isolated high-frequency converter output circuit, a signal controller, electrodes, and a power supply. The output terminal of the power supply is connected to the power converter circuit, which is connected to the electrodes via the energy storage circuit and the isolated high-frequency converter output circuit. The signal controller is connected to the control input terminals of the energy storage circuit and the isolated high-frequency converter output circuit, respectively, and controls the pulse output signal of the isolated high-frequency converter output circuit through the signal controller to discharge the electrodes. This invention induces irreversible electrical breakdown in living cells, improves the uniformity of irreversible point perforation, and reduces muscle contraction caused by nerve stimulation due to electroporation. It can be used for in vivo or ex vivo experimental research on solid tumors.

[0006] However, the performance and automation level of the prior art high-frequency bipolar pulse-based electroporation technology scheme still have room for improvement, and for structures to be ablated at different locations or volumes, it is necessary to perform complex numerical combinations and tedious and extensive tests on a large number of ablation parameters, such as pulse frequency, electric field strength, and ablation time, to obtain the corresponding optimal ablation parameter combination. Obviously, such an operation method is prone to a tedious and time-consuming test and analysis process. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Chinese Patent Application Publication No. 110946642A [Patent Document 2] Chinese Patent Application Publication No. 106877729A Summary of the Invention [Problem to be solved by the invention]

[0008] To solve the above technical problems, the present invention proposes an electroporation ablation performance measurement device using bidirectional high-frequency pulses. Based on bipolar high-frequency pulses, the electroporation ablation system is designed efficiently and automatically. The hardware platform allows for customization of different predictors for different regions and volumes of target structures for ablation. A vast number of ablation parameter combinations, such as the numerical simulation analysis mode in the MATLAB® toolbox, can be used to achieve reliable prediction of each ablation result data. A customized selection mechanism can then be used to obtain optimized ablation parameter combinations for different regions and volumes of target structures for ablation. This allows for rapid provision of optimal low-damage ablation schemes for different regions and volumes of target structures, ensuring ablation effectiveness while minimizing damage to living cells within the structures. [Means for solving the problem]

[0009] Specifically, in a first aspect of the present invention, an electroporation ablation performance measurement device using a high-frequency bidirectional pulse is proposed. The electroporation ablation performance measurement device using a high-frequency bidirectional pulse comprises: a pulse generating mechanism comprising a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate the high frequency bidirectional pulses to a cell suspension of target cell clusters; The target cell cluster, which is a structure composed of multiple living cells from the same site, is flushed twice with 1 to 2 mL of phosphate buffered saline, and then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After digestion for 2 minutes, the target cell cluster is centrifuged in the centrifuge. The supernatant is discarded, and 1640 medium is added to the target cell cluster to grow the target cell cluster at a cell density of 5 x 10. 5 mL -1 a suspension preparation mechanism for preparing the cell suspension; an ablation mechanism for performing an ablation treatment of a predetermined electric field strength on the cell suspension using the high-frequency bidirectional pulse having a target pulse frequency, and obtaining a treated solution after the ablation treatment has reached a predetermined duration; a data measurement device for measuring the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and for determining the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; a network construction device for constructing a deep feedforward network that performs ablation performance prediction using a target pulse frequency used in the ablation treatment using the high-frequency bidirectional pulse, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster as multiple input data of the deep feedforward network, and a cell suppression rate and a low-damage ablation performance level obtained in the ablation treatment as two output data of the deep feedforward network; a sequential learning device connected to the network construction device, for performing one learning operation on the deep feedforward network using a target pulse frequency, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster to be used in each ablation process as multiple input data for the deep feedforward network, and using the cell inhibition rate and low-damage ablation performance level obtained in each ablation process as two output data for the deep feedforward network, and outputting the deep feedforward network after a predetermined number of learning operations as an intelligent predictor; a traversal analysis device connected to the sequential learning device, for traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration for a target cell cluster consisting of a certain total number of living cells at a predetermined site, and for executing the predictive operation of the intelligent predictor for each combination of values, to obtain each cell inhibition rate and each low-damage ablation performance level corresponding to each combination of values, the traversal being based on a numerical simulation process; A best selection analysis mechanism connected to the traversal analysis device for selecting the best ablation performance numerical combination as the optimized ablation numerical combination for a target cell cluster consisting of a certain total number of live cells at a specified site based on each cell inhibition rate corresponding to each numerical combination and each low-damage ablation performance level.

[0010] Specifically, in a second aspect of the present invention, a method for measuring electroporation ablation performance using high-frequency bidirectional pulses is proposed, which comprises: using a pulse generating mechanism comprising a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate high frequency bidirectional pulses to a cell suspension of target cell clusters; The system is equipped with a physiological saline flushing device, a digestion treatment device, a centrifuge, a robot arm, a timing device, a positioning device, a first preparation container, a second preparation container, and a culture preparation device. Target cell clusters, which are structures composed of multiple living cells from the same location, are flushed twice with 1 to 2 mL of phosphate-buffered physiological saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells are centrifuged in a centrifuge, the supernatant is discarded, and 1640 medium is added to culture the target cell clusters at a cell density of 5 × 10 5 mL -1 using a suspension preparation mechanism to prepare a cell suspension of using an ablation mechanism to ablate the cell suspension with a high frequency bidirectional pulse having a target pulse frequency at a predetermined electric field strength, and obtain a treated solution after the ablation has reached a predetermined duration; using a data measurement device to measure the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and to determine the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; Using a network construction device to construct a deep feedforward network for performing ablation performance prediction, the deep feedforward network receives as input data a target pulse frequency to be used in the ablation treatment using high-frequency bidirectional pulses, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster, and the deep feedforward network receives as output data a percentage of cell inhibition obtained in the ablation treatment and a low-damage ablation performance level; using an iterative learning device connected to the network construction device, for performing one learning operation on the deep feedforward network using the target pulse frequency, the predetermined electric field strength, the predetermined duration, and the total number of living cells in the target cell cluster to be used in each ablation treatment as multiple input data of the deep feedforward network, and the cell inhibition rate and the low-damage ablation performance level obtained in each ablation treatment as two output data of the deep feedforward network, and outputting the deep feedforward network after a predetermined number of learning operations as an intelligent predictor; using a traversal analyzer connected to the iterative learning device, for a target cell cluster consisting of a certain total number of living cells at a predetermined site, to traverse each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration, and to perform a predictive operation of an intelligent predictor for each combination of values, to obtain each percentage of cell inhibition and each low-damage ablation performance level corresponding to each combination of values, the traversal being based on a numerical simulation process; A step of using a best selection analysis mechanism connected to the traversal analysis device to select the best ablation performance numerical combination as the optimized ablation numerical combination for a target cell cluster consisting of a certain total number of live cells at a specified site based on each cell inhibition rate corresponding to each numerical combination and each low-damage ablation performance level. [Effects of the Invention]

[0011] From this, it is clear that the present invention has at least the following four important inventive points: (1) A custom-designed ablation treatment system is used, which includes a pulse generating mechanism, a suspension preparation mechanism, and an ablation treatment mechanism, wherein the pulse generating mechanism includes a bipolar pulse generator and a pulse control unit, and the suspension preparation mechanism includes a saline flushing device, a digestion treatment device, a centrifuge, a robot arm, a timing device, a positioning device, a first preparation container, a second preparation container, and a culture preparation device, thereby realizing targeted ablation treatment of a structure consisting of multiple living cells in the same location, thereby improving the performance and automation level of electroporation ablation treatment using high-frequency bidirectional pulses; (2) A deep feedforward network is established for predicting ablation performance, using the target pulse frequency used in the high-frequency bidirectional pulse ablation process, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the structure as multiple input data of the deep feedforward network, and the cell inhibition rate and low-damage ablation performance level obtained in the ablation process as binary output data of the deep feedforward network, and a sequential learning operation is performed on the deep feedforward network to obtain an intelligent predictor that can be used to predict ablation performance; (3) For a structure consisting of a certain total number of living cells at a predetermined location, traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration, and performing a predictive operation of an intelligent predictor for each combination of values ​​to obtain each cell inhibition rate and each low-damage ablation performance level corresponding to each combination of values, and the traversal operation is based on a numerical simulation process, thereby realizing an efficient and reliable replacement of a huge amount of testing; (4) Based on the respective cell inhibition rates and respective low-damage ablation performance levels corresponding to each numerical combination, a numerical combination showing the best ablation processing performance is selected as the optimized ablation numerical combination for a structure consisting of a certain total number of living cells at a specified site. By using a target priority mechanism that determines the numerical combination with the highest cell inhibition rate and the highest low-damage ablation performance level at the same cell inhibition rate as the optimal ablation performance numerical combination, optimized ablation numerical combinations with the best ablation processing performance can be obtained for structures of different types and volumes, thereby avoiding the need for a tedious and time-consuming testing and analysis process.

[0012] In order to clearly describe the embodiments of the present invention or the technical means in the prior art, the accompanying drawings that need to be used to depict the embodiments will be briefly described below. Obviously, the accompanying drawings described below are only some embodiments of the present invention, and those skilled in the art can obtain other accompanying drawings based on these accompanying drawings without any creative activity. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a schematic diagram illustrating the inventive concept of the apparatus and method for measuring electroporation ablation performance using high frequency bidirectional pulses of the present invention. FIG. [Figure 2] 1 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a first embodiment of the present invention. [Figure 3] FIG. 10 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a second embodiment of the present invention. [Figure 4] FIG. 10 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a third embodiment of the present invention. [Figure 5] FIG. 10 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a fourth embodiment of the present invention. [Figure 6] FIG. 10 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a fifth embodiment of the present invention. [Figure 7] 10 is a flowchart showing steps of a method for measuring electroporation ablation performance using high-frequency bidirectional pulses according to a sixth embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0014] Hereinafter, embodiments of the device and method for measuring electroporation ablation performance using high-frequency bidirectional pulses according to the present invention will be described in detail with reference to the accompanying drawings.

[0015] As shown in FIG. 1, the specific inventive concept of the device and method for measuring electroporation ablation performance using high-frequency bidirectional pulses of the present invention is as follows: Step 1: Build a custom-designed ablation system, the ablation system comprising a pulse generating mechanism, a suspension preparation mechanism, and an ablation mechanism, the custom design being used to improve the performance and automation level of electroporation ablation using high frequency bidirectional pulses; Step 2: Construct an intelligent predictor for predicting ablation performance, which takes a target pulse frequency used in the high-frequency bidirectional pulse ablation process, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the structure as multiple input data, and takes the cell inhibition rate and low-damage ablation performance level obtained in the ablation process as two output data, and the intelligent predictor is based on a deep feedforward network and has completed a predetermined number of network learning rounds, thereby ensuring the reliability of the prediction by the intelligent predictor; Step 3: For a structure consisting of a certain total number of living cells at a predetermined site, the intelligent predictor is executed to traverse each combination of the target pulse frequency, the predetermined field strength, and the predetermined duration, and obtain the ablation performance of each prediction. The traversal operation is based on a numerical simulation mode similar to the MATLAB toolbox, Step 4: For a structure consisting of a certain total number of living cells at a specified site, the combination of target pulse frequency, specified electric field strength, and specified duration corresponding to the best ablation performance from each predicted ablation performance is selected as the optimized ablation value combination for the custom structure, effectively replacing the tedious and extensive ablation testing and analysis process.

[0016] The key points of this invention are that the custom-structured ablation processing system optimizes the hardware foundation of the ablation processing, and by constructing an intelligent predictor, provides a reliable simulation model for effective replacement of the subsequent testing and analysis process, and by using traversal processing based on numerical simulation, it is possible to avoid the complicated and time-consuming comparison of ablation parameters, and obtain polynomial optimized ablation parameters corresponding to various structures with rapid speed and reliable accuracy.

[0017] Hereinafter, the present invention will be specifically described with reference to each embodiment of the present invention.

[0018] (First embodiment) FIG. 2 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using high-frequency bidirectional pulses according to a first embodiment of the present invention, the device comprising a pulse generation mechanism, a suspension preparation mechanism, an ablation processing mechanism, a data measurement mechanism, a network construction mechanism, a sequential learning mechanism, a traversal analysis mechanism, and a best selection analysis mechanism; the pulse generating mechanism comprises a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate high frequency bidirectional pulses to the cell suspension of the target cell clusters; Specifically, the specific pulse generation mode of the bipolar pulse generator is to first apply a positive pulse, then apply a negative pulse with the same pulse width after a delay of 2 μs, and then apply a positive pulse after another delay of 2 μs, repeating this process until the total high level time is 100 μs (50 μs for each positive and negative pulse), and then forming a set of pulse trains. This application uses a bipolar pulse format, that is, it is composed of a positive pulse and a negative pulse, and the frequency has the same parameters, so it is defined as a high-frequency bipolar pulse, Specifically, the bipolar pulse generator may be developed in the laboratory itself, and the electrode cup is 4 mm wide. The voltage and current feedback signals are collected by a WavePro 760Zi-A oscilloscope, and a PPE-5 kV high voltage probe and a Pearson 6600 current coil are equipped. The suspension preparation mechanism includes a saline flushing device, a digestion treatment device, a centrifuge, a robot arm, a timing device, a positioning device, a first preparation container, a second preparation container, and a culture preparation device. The target cell clusters are flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution with a mass concentration of 0.5 g / L, and after 2 minutes of digestion, centrifuged in a centrifuge. The supernatant is discarded, and 1640 medium is added to the suspension to obtain a cell density of 5 × 10. 5 mL -1 a cell suspension comprising: a target cell cluster; a structure composed of a plurality of living cells in the same region; The ablation processing mechanism performs ablation processing of a predetermined electric field strength on the cell suspension using a high-frequency bidirectional pulse having a target pulse frequency, and obtains a processed solution after the ablation processing has reached a predetermined duration; the data measurement device is used to measure the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and to determine the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; The network construction device is used to construct a deep feedforward network for performing ablation performance prediction, using a target pulse frequency used in the ablation treatment using high-frequency bidirectional pulses, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster as multiple input data of the deep feedforward network, and a cell suppression rate and a low-damage ablation performance level obtained in the ablation treatment as two output data of the deep feedforward network; For example, the deep feedforward network may include a single input layer, a single output layer, and multiple hidden layers between the single input layer and the single output layer; the number of hidden layers between the single input layer and the single output layer is positively correlated with the total number of live cells in the target cell cluster; The sequential learning device is connected to the network construction device, and is used to perform one learning operation on the deep feedforward network using a target pulse frequency used in each ablation process, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster as multiple input data for the deep feedforward network, and a cell suppression rate and a low-damage ablation performance level obtained in each ablation process as two output data for the deep feedforward network, and to output the deep feedforward network that has undergone a predetermined number of learning operations as an intelligent predictor. a traversal analysis device connected to the sequential learning device, for a target cell cluster consisting of a certain total number of living cells at a predetermined site, traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration, and performing a predictive operation of an intelligent predictor for each combination of values ​​to obtain each cell inhibition rate and each low-damage ablation performance level corresponding to each combination of values, wherein the traversal is based on a numerical simulation process; a best selection analysis mechanism connected to the traversal analysis device, for selecting a best ablation performance value combination as an optimized ablation value combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each value combination; Determining the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells includes taking the sum of the percentage of necrotic cells and the percentage of apoptotic cells as the percentage of cell inhibition obtained by the ablation treatment, dividing the percentage of apoptotic cells by the percentage of cell inhibition to obtain an ablation performance ratio, and determining the low-damage ablation performance level proportional to the ablation performance ratio; Selecting the best ablation performance value combination as an optimized ablation value combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site based on each percentage of cell inhibition and each low-damage ablation performance level corresponding to each value combination includes selecting the value combination with the highest corresponding percentage of cell inhibition as an optimized ablation value combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site; The network construction device, the sequential learning device, the traversal analysis device, and the best selection analysis mechanism can be implemented using different types of SOC chips or CPLD chips.

[0019] Next, a specific configuration of the electroporation ablation performance measurement device using high-frequency bidirectional pulses according to the present invention will be described in detail with reference to each embodiment of the present invention.

[0020] (Second embodiment) 3 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to a second embodiment of the present invention. Unlike the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the first embodiment of the present invention, the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the second embodiment of the present invention further comprises a synchronous drive mechanism connected to the saline flushing device, digestion treatment device, centrifuge, robot arm, timing device and positioning device in the suspension preparation mechanism, The synchronous drive mechanism is used to realize synchronous control of the movements among the saline flushing device, the digestion treatment device, the centrifuge, the robot arm, the timing device and the positioning device.

[0021] (Third embodiment) 4 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to a third embodiment of the present invention. Unlike the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the first embodiment of the present invention, the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the third embodiment of the present invention further comprises a real-time notification mechanism connected to the best selection analysis mechanism for receiving and notifying a combination of optimized ablation values ​​for a target cell cluster consisting of a certain total number of living cells at a predetermined site; Specifically, the instant notification mechanism includes a voice playback unit for announcing an optimized ablation value combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site in a voice playback mode; In particular, the instant notification mechanism includes a screen display unit for notifying, in a screen display mode, a combination of optimized ablation values ​​for a target cell cluster consisting of a certain total number of living cells at a predetermined site.

[0022] (Fourth embodiment) 5 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to a fourth embodiment of the present invention. Unlike the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the first embodiment of the present invention, the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the fourth embodiment of the present invention further comprises an information storage chip connected to the traversal analysis device for storing each predictor parameter of the intelligent predictor, An information storage chip is connected to the traversal analysis device and is used to store each predictor parameter of the intelligent predictor, including that the information storage chip is either a static storage chip, a TF memory, or an MMC memory.

[0023] (Fifth embodiment) 6 is a schematic diagram showing the internal structure of an electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to a fifth embodiment of the present invention. Unlike the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the first embodiment of the present invention, the electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to the fifth embodiment of the present invention further includes parallel communication interfaces connected to the network construction device, the traversal analysis device, and the best selection analysis mechanism, respectively; the parallel communication interface is used to realize parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism; The parallel communication interface being used to realize parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism includes that the realized parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism is 16-bit parallel data communication; The parallel communication interface being used to realize parallel data communication between the network construction device, the traversal analysis device and the best selection analysis mechanism includes using a serial control interface to connect to the parallel communication interface and to control the real-time communication mode of the parallel communication interface.

[0024] In any one of the embodiments of the present invention, the electroporation ablation performance measurement device using high frequency bidirectional pulses is The network construction device for constructing a deep feedforward network for performing ablation performance prediction, using a target pulse frequency used in ablation treatment using high-frequency bidirectional pulses, a predetermined electric field strength, a predetermined duration, and the total number of living cells in a target cell cluster as multiple input data of the deep feedforward network, and a percentage of cell inhibition obtained in the ablation treatment and a low-damage ablation performance level as two output data of the deep feedforward network, includes a reference value of the target pulse frequency being 71.4 kHz, a reference value of the predetermined electric field strength being 0.2 MV / m, and a reference value of the predetermined duration being 12 hours; Controlling the pulse generation mode of the bipolar pulse generator to generate high frequency bidirectional pulses to the cell suspension of the target cell clusters includes having a frequency of the high frequency bidirectional pulses greater than or equal to 71.4 kHz.

[0025] In any one of the embodiments of the present invention, the electroporation ablation performance measurement device using high frequency bidirectional pulses is The target cell clusters were flushed twice with 1-2 mL of phosphate-buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells to a cell density of 5 × 10 5 mL -1 a suspension preparation mechanism for preparing the cell suspension, the mechanism comprising the saline flushing device for flushing the target cell clusters twice with 1 to 2 mL of phosphate buffered saline; The target cell clusters were flushed twice with 1-2 mL of phosphate-buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells to a cell density of 5 × 10 5 mL -1 a suspension preparation mechanism for preparing a cell suspension, the mechanism being connected to the timer and including the digestion treatment device for digesting the target cell clusters after flushing with a pancreatic enzyme solution having a mass concentration of 0.5 g / L for 2 minutes, the timer being used to time the digestion treatment device; The target cell clusters were flushed twice with 1-2 mL of phosphate-buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells to a cell density of 5 × 10 5 mL -1 The suspension preparation mechanism for preparing the cell suspension further comprises the centrifuge for centrifuging the solution after digestion for 2 minutes.

[0026] In any one of the embodiments of the present invention, the electroporation ablation performance measurement device using high frequency bidirectional pulses is The target cell clusters were flushed twice with 1-2 mL of phosphate-buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells to a cell density of 5 × 10 5 mL -1a positioning device connected to the robot arm for providing positioning data for the robot arm; and a robot arm for clamping a first preparation container storing a solution after 2 minutes of digestion, a second preparation container storing a solution from which the supernatant has been discarded, or target cell clusters after flushing, The target cell clusters were flushed twice with 1-2 mL of phosphate-buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells to a cell density of 5 × 10 5 mL -1 The suspension preparation mechanism for preparing the cell suspension of 1640 medium was added to the second preparation vessel containing the discarded supernatant solution to obtain a cell density of 5 × 10 5 mL -1 The cell culture preparation device is provided for preparing a cell suspension of the above.

[0027] (Sixth embodiment) 7 is a flowchart showing steps of a method for measuring electroporation ablation performance using high-frequency bidirectional pulses according to a sixth embodiment of the present invention. The method for measuring electroporation ablation performance using high-frequency bidirectional pulses comprises the following steps: using a pulse generating mechanism comprising a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate high frequency bidirectional pulses to a cell suspension of target cell clusters; Specifically, the specific pulse generation mode of the bipolar pulse generator is to first apply a positive pulse, then apply a negative pulse with the same pulse width after a delay of 2 μs, and then apply a positive pulse after another delay of 2 μs, repeating this process until the total high level time is 100 μs (50 μs for each positive and negative pulse), and then forming a set of pulse trains. This application uses a bipolar pulse format, that is, it is composed of a positive pulse and a negative pulse, and the frequency has the same parameters, so it is defined as a high-frequency bipolar pulse, Specifically, the bipolar pulse generator may be developed in the laboratory itself, and the electrode cup is 4 mm wide. The voltage and current feedback signals are collected by a WavePro 760Zi-A oscilloscope, and a PPE-5 kV high voltage probe and a Pearson 6600 current coil are equipped. The system is equipped with a physiological saline flushing device, a digestion treatment device, a centrifuge, a robot arm, a timing device, a positioning device, a first preparation container, a second preparation container, and a culture preparation device. Target cell clusters, which are structures composed of multiple living cells from the same location, are flushed twice with 1 to 2 mL of phosphate-buffered physiological saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells are centrifuged in a centrifuge, the supernatant is discarded, and 1640 medium is added to culture the target cell clusters at a cell density of 5 × 10 5 mL -1 using a suspension preparation mechanism to prepare a cell suspension of using an ablation mechanism to ablate the cell suspension with a high frequency bidirectional pulse having a target pulse frequency at a predetermined electric field strength, and obtain a treated solution after the ablation has reached a predetermined duration; using a data measurement device to measure the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and to determine the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; Using a network construction device to construct a deep feedforward network for performing ablation performance prediction, the deep feedforward network receives as input data a target pulse frequency to be used in the ablation treatment using high-frequency bidirectional pulses, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster, and the deep feedforward network receives as output data a percentage of cell inhibition obtained in the ablation treatment and a low-damage ablation performance level; For example, the deep feedforward network may include a single input layer, a single output layer, and multiple hidden layers between the single input layer and the single output layer; the number of hidden layers between the single input layer and the single output layer is positively correlated with the total number of live cells in the target cell cluster; using an iterative learning device connected to the network construction device, for performing one learning operation on the deep feedforward network using the target pulse frequency, the predetermined electric field strength, the predetermined duration, and the total number of living cells in the target cell cluster to be used in each ablation treatment as multiple input data of the deep feedforward network, and the cell inhibition rate and the low-damage ablation performance level obtained in each ablation treatment as two output data of the deep feedforward network, and outputting the deep feedforward network after a predetermined number of learning operations as an intelligent predictor; a traversal analysis device connected to the iterative learning device, for a target cell cluster consisting of a certain total number of living cells at a predetermined site, traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration, and performing a predictive operation of an intelligent predictor for each combination of values ​​to obtain each percentage of cell inhibition and each low-damage ablation performance level corresponding to each combination of values, the traversal being based on a numerical simulation process; using a best selection analysis mechanism connected to the traversal analysis device for selecting a best ablation performance value combination as an optimized ablation value combination for a target cell cluster consisting of a certain total number of live cells at a predetermined site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each value combination; Determining the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells includes taking the sum of the percentage of necrotic cells and the percentage of apoptotic cells as the percentage of cell inhibition obtained by the ablation treatment, dividing the percentage of apoptotic cells by the percentage of cell inhibition to obtain an ablation performance ratio, and determining the low-damage ablation performance level proportional to the ablation performance ratio; Selecting the best ablation performance value combination as the optimized ablation value combination for a target cell cluster consisting of a certain total number of viable cells at a given site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each value combination. selecting the value combination with the highest corresponding percentage of cell inhibition as the optimized ablation value combination for a target cell cluster consisting of a certain total number of viable cells at a predetermined site; The network construction device, the sequential learning device, the traversal analysis device, and the best selection analysis mechanism can be implemented using different types of SOC chips or CPLD chips.

[0028] In addition, the present invention can highlight the significant technical advances of the present invention by citing the following technical content: Selecting the best ablation performance numerical combination as an optimized ablation numerical combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each numerical combination further includes, when the cell inhibition rate corresponding to two or more numerical combinations is the highest, selecting the numerical combination with the highest corresponding low-damage ablation performance level as the optimized ablation numerical combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site; For a target cell cluster consisting of a certain total number of living cells at a certain location, traversing each numerical combination of a target pulse frequency, a predetermined electric field strength, and a predetermined duration, executing a predictive operation of an intelligent predictor for each numerical combination, and obtaining each cell inhibition rate and each low-damage ablation performance level corresponding to each numerical combination, in the predictive operation of the intelligent predictor for each numerical combination, the target pulse frequency, the predetermined electric field strength, the predetermined duration, and a certain total number are used as multiple input data for the intelligent predictor, executing the intelligent predictor, and obtaining the cell inhibition rate and low-damage ablation performance level output from the intelligent predictor, which are the predicted results of the predictive operation of the intelligent predictor.

[0029] As will be explained later, in this specification, relational terms such as "first," "second," etc. are used to distinguish one entity or operation from another without necessarily requiring or implying the existence of any actual relationship or order between those entities or operations. Furthermore, the terms "comprise," "comprises," or any other variant thereof are intended to cover a non-exclusive inclusion, whereby a process, method, article, or device comprising a set of elements includes not only those elements but also other elements not expressly listed or inherent in such process, method, article, or device. Absent more limitations, elements qualified by the phrase "comprises a..." do not exclude the inclusion of other identical elements in the process, method, article, or device that includes the element.

[0030] Each embodiment of this specification will be described in a progressive manner, and each embodiment will mainly describe the differences from other embodiments, and similar or similar parts between the embodiments may be referred to each other. In particular, the device / electronic device / computer-readable recording medium / computer program product embodiments are basically similar to the method embodiments, so the description thereof will be relatively simple, and the relevant parts may be referred to the description of the method embodiments.

[0031] The above description is only a preferred embodiment of the present invention, and is not intended to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention are also included in the protection scope of the present invention.

Claims

1. An apparatus for measuring electroporation ablation performance using high frequency bidirectional pulses, a pulse generating mechanism comprising a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate the high frequency bidirectional pulses to a cell suspension of target cell clusters; The target cell cluster, which is a structure composed of multiple living cells from the same site, is flushed twice with 1 to 2 mL of phosphate buffered saline, and then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After digestion for 2 minutes, the target cell cluster is centrifuged in the centrifuge. The supernatant is discarded, and 1640 medium is added to the target cell cluster to grow the target cell cluster at a cell density of 5 x 10. ５ mL －１ a suspension preparation mechanism for preparing the cell suspension; an ablation mechanism for performing an ablation treatment of a predetermined electric field strength on the cell suspension using the high-frequency bidirectional pulse having a target pulse frequency, and obtaining a treated solution after the ablation treatment has reached a predetermined duration; a data measurement device for measuring the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and for determining the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; a network construction device for constructing a deep feedforward network that performs ablation performance prediction using a target pulse frequency used in the ablation treatment using the high-frequency bidirectional pulse, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster as multiple input data of the deep feedforward network, and a cell suppression rate and a low-damage ablation performance level obtained in the ablation treatment as two output data of the deep feedforward network; a sequential learning device connected to the network construction device, for performing one learning operation on the deep feedforward network using a target pulse frequency, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster to be used in each ablation process as multiple input data for the deep feedforward network, and using the cell inhibition rate and low-damage ablation performance level obtained in each ablation process as two output data for the deep feedforward network, and outputting the deep feedforward network after a predetermined number of learning operations as an intelligent predictor; a traversal analysis device connected to the sequential learning device, for traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration for a target cell cluster consisting of a certain total number of living cells at a predetermined site, and for executing the predictive operation of the intelligent predictor for each combination of values, to obtain each cell inhibition rate and each low-damage ablation performance level corresponding to each combination of values, the traversal being based on a numerical simulation process; a best selection analysis mechanism connected to the traversal analysis device for selecting a best ablation performance value combination as an optimized ablation value combination for a target cell cluster consisting of a certain total number of living cells at a predetermined site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each value combination; An electroporation ablation performance measurement device using high-frequency bidirectional pulses, comprising:

2. Determining the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells includes taking the sum of the percentage of necrotic cells and the percentage of apoptotic cells as the percentage of cell inhibition obtained by the ablation treatment, dividing the percentage of apoptotic cells by the percentage of cell inhibition to obtain an ablation performance ratio, and determining the low-damage ablation performance level proportional to the ablation performance ratio; Selecting the best ablation performance value combination as the optimized ablation performance value for a target cell cluster consisting of a certain total number of living cells at a predetermined site based on each percentage of cell inhibition and each low-damage ablation performance level corresponding to each value combination includes selecting the value combination with the highest corresponding percentage of cell inhibition as the optimized ablation performance value for a target cell cluster consisting of a certain total number of living cells at a predetermined site.

2. The electroporation ablation performance measurement device using high-frequency bidirectional pulses according to claim 1.

3. the suspension preparation mechanism further comprises a synchronous drive mechanism connected to the saline flushing device, the digestion treatment device, the centrifuge, the robot arm, the timing device, and the positioning device, respectively, for realizing synchronous control of the operations among the saline flushing device, the digestion treatment device, the centrifuge, the robot arm, the timing device, and the positioning device; 3. The electroporation ablation performance measurement device using high frequency bidirectional pulses according to claim 2.

4. and a real-time notification mechanism connected to the best selection analysis mechanism for receiving and notifying a combination of optimized ablation values ​​for a target cell cluster consisting of a certain total number of living cells at a predetermined site.

3. The electroporation ablation performance measurement device using high frequency bidirectional pulses according to claim 2.

5. further comprising an information storage chip connected to the traversal analyzer for storing predictor parameters of each of the intelligent predictors; 3. The electroporation ablation performance measurement device using high frequency bidirectional pulses according to claim 2.

6. a parallel communication interface connected to the network construction device, the traversal analysis device, and the best selection analysis mechanism, the parallel communication interface is used to realize parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism; The parallel communication interface being used to realize parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism includes that the realized parallel data communication between the network construction device, the traversal analysis device, and the best selection analysis mechanism is 16-bit parallel data communication.

3. The electroporation ablation performance measurement device using high frequency bidirectional pulses according to claim 2.

7. The network construction device for constructing a deep feedforward network for performing ablation performance prediction using a target pulse frequency, a predetermined electric field strength, a predetermined duration, and the total number of living cells in a target cell cluster as a plurality of input data of the deep feedforward network, and a cell inhibition rate and a low-damage ablation performance level obtained in the ablation process as two output data of the deep feedforward network, includes a reference value of the target pulse frequency of 71.4 kHz, a reference value of the predetermined electric field strength of 0.2 MV / m, and a reference value of the predetermined duration of 12 hours; controlling the pulse generation mode of the bipolar pulse generator to generate the high frequency bidirectional pulses to the cell suspension of the target cell clusters includes the frequency of the high frequency bidirectional pulses being greater than or equal to 71.4 kHz; 7. The electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to claim 2.

8. The target cell clusters were flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells at a cell density of 5 × 10 ５ mL －１ a suspension preparation mechanism for preparing a cell suspension comprising the saline flushing device for flushing the target cell clusters twice with 1 to 2 mL of phosphate buffered saline; The target cell clusters were flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells at a cell density of 5 × 10 ５ mL －１ a digestion treatment device connected to the timer and configured to digest target cell clusters after flushing with a pancreatic enzyme solution having a mass concentration of 0.5 g / L for 2 minutes, the digestion treatment device being used to time the digestion treatment device; The target cell clusters were flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells at a cell density of 5 × 10 ５ mL －１ The suspension preparation mechanism for preparing the cell suspension further comprises the centrifuge for centrifuging the solution after digestion for 2 minutes.

7. The electroporation ablation performance measurement device using a high-frequency bidirectional pulse according to claim 2.

9. The target cell clusters were flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells at a cell density of 5 × 10 ５ mL －１ a positioning device connected to the robot arm for providing positioning data for the robot arm; and a robot arm for clamping a first preparation container storing a solution after 2 minutes of digestion, a second preparation container storing a solution from which the supernatant has been discarded, or target cell clusters after flushing, The target cell clusters were flushed twice with 1-2 mL of phosphate buffered saline, then digested with a pancreatic enzyme solution at a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells were centrifuged in a centrifuge, the supernatant was discarded, and 1640 medium was added to the cells at a cell density of 5 × 10 ５ mL －１ The suspension preparation mechanism for preparing the cell suspension of 1640 medium was added to the second preparation vessel containing the solution from which the supernatant had been discarded, and the cell density was set to 5×10 ５ mL －１ The culture preparation device is provided for preparing a cell suspension of 9. The electroporation ablation performance measurement device using high frequency bidirectional pulses according to claim 8.

10. A method for measuring electroporation ablation performance using high frequency bidirectional pulses, comprising: using a pulse generating mechanism comprising a bipolar pulse generator and a pulse control unit connected to the bipolar pulse generator for controlling a pulse generating mode of the bipolar pulse generator to generate high frequency bidirectional pulses to a cell suspension of target cell clusters; The apparatus is equipped with a physiological saline flushing device, a digestion treatment device, a centrifuge, a robot arm, a timing device, a positioning device, a first preparation container, a second preparation container, and a culture preparation device. Target cell clusters, which are structures composed of multiple living cells from the same site, are flushed twice with 1 to 2 mL of phosphate buffered physiological saline, then digested with a pancreatic enzyme solution with a mass concentration of 0.5 g / L. After 2 minutes of digestion, the cells are centrifuged in a centrifuge, the supernatant is discarded, and 1640 medium is added to culture the target cell clusters at a cell density of 5 x 10. ５ mL －１ using a suspension preparation mechanism to prepare a cell suspension of using an ablation mechanism to ablate the cell suspension with a high frequency bidirectional pulse having a target pulse frequency at a predetermined electric field strength, and obtain a treated solution after the ablation has reached a predetermined duration; using a data measurement device to measure the total number of viable cells, the percentage of necrotic cells, and the percentage of apoptotic cells remaining in the treatment solution after the ablation treatment has reached a predetermined duration, and to determine the percentage of cell inhibition and the low-damage ablation performance level obtained by the ablation treatment based on the percentage of necrotic cells and the percentage of apoptotic cells; using a network construction device to construct a deep feedforward network for performing ablation performance prediction, using a target pulse frequency used in the high-frequency bidirectional pulse ablation treatment, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster as multiple input data of the deep feedforward network, and a percentage of cell inhibition obtained in the ablation treatment and a low-damage ablation performance level as two output data of the deep feedforward network; using an iterative learning device connected to the network construction device, for performing one learning operation on the deep feedforward network using the target pulse frequency, a predetermined electric field strength, a predetermined duration, and the total number of living cells in the target cell cluster used in each ablation process as multiple input data of the deep feedforward network, and the cell inhibition rate and the low-damage ablation performance level obtained in each ablation process as two output data of the deep feedforward network, and outputting the deep feedforward network after a predetermined number of learning operations as an intelligent predictor; a traversal analysis device connected to the iterative learning device, for a target cell cluster consisting of a certain total number of living cells at a predetermined site, traversing each combination of values ​​of a target pulse frequency, a predetermined field strength, and a predetermined duration, and performing a predictive operation of an intelligent predictor for each combination of values ​​to obtain each percentage of cell inhibition and each low-damage ablation performance level corresponding to each combination of values, the traversal being based on a numerical simulation process; using a best selection analysis mechanism connected to the traversal analysis device for selecting the best ablation performance value combination as an optimized ablation value combination for a target cell cluster consisting of a certain total number of live cells at a predetermined site based on each cell inhibition rate and each low-damage ablation performance level corresponding to each value combination; A method for measuring electroporation ablation performance using high-frequency bidirectional pulses, comprising:

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