Apparatus and method for electroporation

Abstract

An apparatus and method for electroporation of cells is disclosed. The apparatus includes a memory for storing instructions, a high-voltage module for applying a voltage to cells, a low-voltage module including a user interface, and a system board connected to the high-voltage module, the low-voltage module, and a single-board computer, the single-board computer including at least one processor for controlling the high-voltage module and the low-voltage module. The system board includes an emergency stop button and electronic circuitry. The electronic circuitry receives a signal from the emergency stop button, latches the signal from the emergency stop button, blocks a logic signal that controls the voltage provided to the meter output, stops the voltage applied to the cells, and disables the at least one processor from providing commands to the high-voltage module.

Description

[Technical Field]

[0001] Priority This application claims priority to U.S. Provisional Application No. 63 / 404,157, filed September 6, 2022, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates generally to devices, structures, and methods for electroporation. In particular, embodiments of the present disclosure relate to an inventive and unconventional device and method for providing electroporation of cells in a cell suspension from a fluid sample, the device integrating a display and touch screen, a computer, application of high voltage to cells in a cell suspension from a fluid sample, and a non-software-based emergency stop function. [Background technology]

[0003] Electroporation systems can apply a voltage to cells in a fluid cell suspension via an electric field to increase the permeability of the cell membrane, allowing chemicals, drugs, or nucleic acids (DNA or RNA) to be introduced into the cells. The application of the voltage can result in an electric field according to a pulse duration schedule or protocol. Electroporation systems can be used in medical, testing, and microbiology applications.

[0004] Current electroporation systems may be standalone systems designed only for transfection and may require the purchase of a separate desktop or laptop computer to process results from the current electroporation system. For example, current electroporation systems may only provide raw data to operators or users, and operators or users of current electroporation systems may need to purchase an additional desktop or laptop computer to analyze and generate plots from the current electroporation system. Non-integrated electroporation systems that require the purchase of a separate desktop or laptop computer may additionally cause compatibility issues between the desktop or laptop computer's operating system and the electroporation system. For example, current electroporation systems may provide operators or users with separate software to install on the desktop or laptop computer to process raw data generated by the current electroporation system, and the software may help generate, identify, and analyze plots of transfected cells in a fluid cell suspension. In other examples, the software provided for installation on the desktop or laptop computer may be incompatible with or not up-to-date with the existing operating system, which may force operators or users to maintain older versions of the desktop or laptop computer's operating system. Additionally, current electroporation systems may require the use of a separate storage medium to receive the results of the transfection process in the form of a file containing raw data, which may be used in a desktop or laptop computer to process the file for results. The use of a separate storage medium may also require the use of software to process the results in the file.

[0005] The lack of integration of computer functions in current electroporation systems significantly increases costs to operators or users, as they may be forced to purchase a standalone electroporation system and a desktop or laptop computer and incur the cost of converting the desktop or laptop computer's current operating system to an older operating system that is compatible with the software provided with the electroporation system. Furthermore, the lack of integration of computer functions can increase the difficulty of operating these systems, as operators or users may have to rely on the system's operating manual to understand the function of outdated mechanical buttons or switches. For example, current electroporation systems may not utilize a display screen to guide the operator or user through the transfection process or provide the status of the transfection process. For example, current electroporation system manufacturers may rely on standalone software installed on the desktop or laptop computer to provide the operator or user with electroporation status, but these statuses may be delayed due to the lack of integration between the electroporation system, the desktop or laptop computer, and the software.

[0006] Furthermore, current non-integrated electroporation systems with desktop or laptop computers may not provide safety features that allow the operator or user to immediately terminate the transfection process if the operator or user discovers a dangerous condition due to a malfunctioning electroporation system, an incorrect fluid sample ready to be processed, a flammable fluid sample, or the spillage of a conductive sample into or out of the electroporation system, which may create a dangerous situation for the operator or user. For example, the operator or user may be forced to unplug all power sources to the electroporation system or disconnect the desktop or laptop computer from the electroporation system, which may create a very dangerous situation due to the possibility of electrocution or fire. While non-integrated electroporation systems may allow the operator or user to use commands on the desktop or laptop computer, such as the space bar on the keyboard, this implementation of a non-integrated electroporation system using a desktop or laptop computer may not be instantaneous, as the operator may have to go through a series of menus before the software recognizes that using the space bar on the keyboard may mean stopping the electroporation process. Furthermore, the application of the space bar to terminate the electroporation process may be software dependent, which may be limited by errors, delays, and logical sequence steps that may increase the time to execute a shutdown of the electroporation system. In addition, software-based space bar applications to terminate the electroporation process may be susceptible to errors in communication between the desktop or laptop computer and the current electroporation system.

[0007] The lack of a means to immediately terminate the transfection process increases costs to the operator and user due to lost electroporation systems, lost desktop or laptop computers, lost fluid samples, or serious injury to the operator and user. Summary of the Invention [Problem to be solved by the invention]

[0008] Therefore, there is a need for improved devices and methods for electroporation of fluids that integrate computer functionality, apply higher electric field strengths (volts per meter), accommodate larger volumes of fluid up to one liter (1 L), accommodate larger culture sizes, and incorporate a non-software-based emergency stop function to immediately terminate the electroporation process to protect the fluid sample, the electroporation device, and the operator and user. [Means for solving the problem]

[0009] One aspect of the present disclosure is directed to an apparatus for electroporation of cells in a cell suspension in a fluid. The apparatus may include a memory for storing instructions, a high-voltage module including an instrument output configured to apply a voltage to cells in the cell suspension in a fluid, a low-voltage module including a user interface device, a system board connected to the high-voltage module, the low-voltage module, and a single-board computer, the single-board computer including at least one processor configured to (i) send instructions to control the high-voltage module and the low-voltage module, and (ii) receive instructions from the user interface device to adjust the voltage applied to cells in the cell suspension in a fluid by the instrument output, and an emergency stop button coupled to the system board. The system board may further include electronic circuitry that may be configured to execute instructions for performing steps including receiving a signal from the emergency stop button to stop the voltage applied by the meter output to the cells in the cell suspension in the fluid, latching the signal from the emergency stop button, blocking a logic signal that controls the voltage supply to the meter output, terminating the voltage applied by the meter output to the cells in the suspension in the fluid, and disabling the at least one processor from providing commands to the high voltage module.

[0010] Another aspect of the present disclosure is directed to a method for performing safety steps to terminate electroporation of cells in a cell suspension in a fluid, the method may include receiving a signal from an emergency stop button to terminate a voltage applied to the cells in the cell suspension in the fluid by a meter output, latching the signal from the emergency stop button, blocking a logic signal that controls the voltage supply to the meter output, terminating the voltage applied to the cells in the cell suspension in the fluid by the meter output, and disabling at least one processor from providing commands to a high voltage module.

[0011] Yet another aspect of the present disclosure is directed to an apparatus for electroporation of cells in a cell suspension in a fluid. The apparatus may include a memory for storing instructions, a high-voltage module including an instrument output configured to apply a voltage to cells in the cell suspension in a fluid, a low-voltage module including a user interface device, a system board connected to the high-voltage module, the low-voltage module, and a single-board computer, the single-board computer including at least one processor configured to (i) send instructions to control the high-voltage module and the low-voltage module, and (ii) receive instructions from the user interface device to adjust the voltage applied to the cells in the cell suspension in a fluid by the instrument output, an emergency stop button coupled to the system board, and a D-flip-flop within the system board, the D-flip-flop may have a preset input pin connected to the emergency stop button, an output pin and a clock input pin connected to the low-voltage module, and an inverted output pin connected to the high-voltage module. A D flip-flop in the system board may be configured to execute instructions to perform steps including receiving a signal from the emergency stop button to stop the voltage applied to the cells in the cell suspension in the fluid by the meter output; latching the signal from the emergency stop button; blocking a logic signal that controls the voltage supply to the meter output; terminating the voltage applied to the cells in the cell suspension in the fluid by the meter output; and disabling at least one processor from providing commands to the high voltage module.

[0012] Other systems, devices, and methods are also discussed herein. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 is a schematic block diagram illustrating an exemplary embodiment of an integrated electroporation device having a non-software-based emergency stop, consistent with disclosed embodiments. [Figure 2]FIG. 1 is a schematic block diagram illustrating an example non-software based emergency stop function consistent with disclosed embodiments. [Figure 3] FIG. 10 is a diagram of a display and touch screen graphical user interface for resetting an emergency stop signal, consistent with a disclosed embodiment. [Figure 4] 1A-1C are exemplary diagrams of an integrated electroporation system and its components in static and flow electroporation configurations consistent with disclosed embodiments. [Figure 5] FIG. 10 is an exemplary diagram of a processing assembly interfacing with a meter output in a static electroporation configuration, consistent with disclosed embodiments. [Figure 6] FIG. 10 is an exemplary diagram of a processing assembly interfacing with an instrument output in a flow electroporation configuration, consistent with disclosed embodiments. [Figure 7] FIG. 1 is an exemplary diagram of an electroporation system having low-voltage component packaging and a display and touch screen consistent with disclosed embodiments. [Figure 8] FIG. 1 is an illustrative block diagram of an exemplary method for electroporation of cells in a cell suspension in a fluid, consistent with some embodiments of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0014] The following detailed description refers to the accompanying drawings. Wherever possible, the same reference numbers are used in the drawings and the following description to refer to the same or similar parts. Several exemplary embodiments are described herein; however, modifications, adaptations, and other implementations are possible. For example, substitutions, additions, or modifications may be made to the components and steps illustrated in the drawings, and the exemplary methods described herein may be modified by substituting, rearranging, deleting, or adding steps to the disclosed methods. Therefore, the following detailed description is not limited to the disclosed embodiments and examples. Instead, the appropriate scope of the present invention is defined by the appended claims.

[0015] Embodiments of the present disclosure are directed to electroporation devices and methods configured to apply a maximum voltage to cells in a cell suspension in a fluid sample to increase transfection yield by increasing the permeability of the cell membrane for the introduction of chemical or biological samples. Furthermore, embodiments of the present disclosure are directed to electroporation devices and methods configured to transfect cells in a cell suspension for a volume of fluid, e.g., 1 L, containing a culture size of, e.g., 200 billion cells. Furthermore, embodiments of the present disclosure are directed to electroporation devices and methods configured to integrate computer functions, including a display and touchscreen, for ease of use, without the need for a standalone desktop or laptop computer. Furthermore, embodiments of the present disclosure are directed to electroporation devices and methods configured to include emergency and safety stop functions that provide for immediate halt of the electroporation process, rather than being software-based, to reduce cell destruction, device destruction, and operator or user safety.

[0016] FIG. 1 is a schematic block diagram illustrating an exemplary embodiment of an integrated electroporation device with a non-software-based emergency stop consistent with disclosed embodiments. As shown in FIG. 1, system 100 may include one or more devices (components) that may comprise system 100. System 100 may include a single-board computer (SBC) 102 (referred to herein as SBC 102), which may include one or more memory storage devices (not shown in FIG. 1), one or more processors 103 (referred to herein as processor 103), a system board (SB) 104 (referred to herein as SB 104), a high-voltage (HV) module 106 (referred to herein as HV module 106), and a low-voltage (LV) module 108 (referred to herein as LV module 108). Processor 103 and / or SBC 102 may control, manage, and / or collect data within system 100. Processor 103 may be an application-specific integrated circuit (ASIC) or a general-purpose processor. Processor 103 may include two or more processors. For example, the processors may be arranged in parallel, serial, or both to process all or a portion of the computer instructions to be processed. In one embodiment, the processor 103 may reside within the SBC 102 and / or in a dedicated circuit board separate from the SBC 102. The SB 104, the HV module 106, and the LV module 108 may each include components. The SB 104 may be connected to the SB 102 and the processor 103, the HV module 106, and the LV module 108. The processor 103 and / or the SBC 102 may control, manage, and / or collect data from the HV module 106 and the LV module 108 (including their components) via communication with the SB 104. In one embodiment, the processor 103 and / or the SBC 102 may be connected to the HV module 106 and its components and the LV module 108 and its components.A power entry module (PEM) 110 (referred to herein as PEM 110), which may be a further included component of system 100, may generate a 24-volt (24V) direct current (DC) voltage rail to power SBC 102, processor 103, SB 104, LV module 108, and HV module 106. SB 104 may connect PEM 110 to SBC 102 and its components, processor 103, HV module 106 and its components, and LV module 108 and its components. Processor 103 and / or SBC 102 may manage and / or control the voltage applied to components within HV module 106 and / or LV module 108. SB 104 may gate the 24V DC voltage rail from PEM 110 using one or more DC relays (not shown in FIG. 1 ) to power components within SB 104 and HV module 106. Similarly, the LV module 108 may include one or more DC relays (not shown in FIG. 1 ) that gate a 24V DC voltage rail from the PEM 110 to provide power to the SBC 102, the processor 103, and components internal to the LV module 108. In another component, the SB 104 may include one or more DC relays that gate a 24V DC voltage rail from the PEM 110 to provide power to its components and to the HV module 106, the LV module 108, the processor 103, and components within the SBC 102. In yet another embodiment, the PEM 110 may include one or more DC relays that gate a 24V DC voltage rail to provide power to the SBC 102 and its components, the processor 103, the SB 104 and its components, the HV module 106 and its components, and the LV module 108 and its components. In yet another embodiment, the PEM 110 may be directly connected to one or more DC relays or power sources that gate a 24V DC voltage rail to power the SBC 102 and its components, the processor 103, the SB 104 and its components, the HV module 106 and its components, and the LV module 108 and its components.

[0017] The HV module 106 may include a high voltage board 112 (referred to herein as HV board 112), an isolated low voltage (LV) DC power supply 114 (referred to herein as LV DC power supply 114), a high voltage power supply 116 (referred to herein as HV power supply 116), at least one capacitor bank 118 including one or more capacitors, a high voltage switch 119 (referred to herein as HV switch 119), a pulse modulator (PM) board 120 (referred to herein as PM board 120), an instrument output 122, a processing assembly (PA) 124 (referred to herein as PA 124), and a calibration port 126, all of which constitute components of the HV module 106.

[0018] The HV board 112 may gate the isolated LV DC power supply 114 from the PEM 110 using a DC relay (not shown in FIG. 1 ) to provide power to the HV power supply 116. The PEM 110 may provide power to the isolated LV DC power supply 114 to provide power to the HV power supply 116. The HV board 112 may manage and / or adjust the amount of power required to the HV power supply 116 based on the required power demand dictated by the processor 103 and / or the SBC 102. The processor 103 and / or the SBC 102 may be connected to the HV board 112 via the SB 104. The HV power supply 116 may then charge at least one capacitor bank 118 to maintain or quickly generate the amount of voltage required for electroporation. The required voltage may range between -1000 volts and +1000 volts, and the required voltage may be ±600 volts, ±700 volts, ±800 volts, ±900 volts, ±1000 volts, and / or greater. The at least one capacitor bank 118 may include one or more capacitors to generate the required voltage. In one embodiment, the at least one capacitor bank 118 may include four capacitors capable of generating the required voltage. In another embodiment, the at least one capacitor bank may include one or more capacitors to generate the required voltage.

[0019] The PM board 120 may generate positive and / or negative pulse profiles or waves that can be implemented by the HV switch 119 to the instrument output 122. The HV power supply 116 may draw power from the LV DC power supply 114 to charge at least one capacitor bank 118 to energize the HV switch 119. The PM board 120 may generate positive and / or negative pulse profiles or waves to the HV switch 119. The positive or negative pulse profiles or waves from the SBC 102, processor 103, SB 104, and / or PM board 120 may be based on cells in a fluidic cell suspension contained within a chamber of the PA 124 that the SBC 102 may identify. The PM board 120 may implement or apply a pulse width wave generated by the SBC 102 and / or processor 103 that corresponds to, results in, and / or is equal to the required amount of voltage to be applied to the instrument output 122 via the HV switch 119. In another embodiment, the PM board 120 may generate a pulse width wave. In yet another embodiment, the processor 103 may generate a pulse width wave. In one embodiment, the PM board 120, the SBC 102, and / or the processor 103 may generate a pulse width. The voltage at the meter output 122 is applied to a chamber containing cells in a cell suspension contained in the fluid sample in the PA 124. The pulse width wave may be a combination of a positive pulse profile or wave and a negative pulse profile or wave, which may correspond to, result in, and / or be equal to the required amount of voltage to be applied to the chamber containing cells in a cell suspension from the fluid sample in the PA 124 via the HV switch 119 and the meter output 122. In another embodiment, the PM board 120 may generate positive and negative pulse profiles or waves to implement a voltage that corresponds to, result in, and / or is equal to the required amount of voltage to be applied by the HV switch 119 to the chamber containing cells in a cell suspension in the fluid sample in the PA 124 via the meter output 122.

[0020] The processor 103 and / or SBC 102 may apply a protocol that may include the timing or duration and precise amplitude and profile of the pulse width wave. The processor 103 and / or SBC 102 may command, manage, and / or regulate the HV board 112, the HV switch 119, the PM board 120, and the meter output 122. In one embodiment, the processor 103 and / or SBC 102 may also detect that there may be no fluid in the chamber of the PA 124 or that there may be no PA assembly connected to the meter output 122.

[0021] The meter output 122 may be configured to provide two modes of operation: static electroporation and flow electroporation. The meter output 122 may have a knob or other device that an operator or user can use to set whether the system 100 operates as static electroporation (small-scale) or flow electroporation (large-scale). The processor 103 and / or SBC 102 may detect the position of the knob or other device on the meter output 122 based on the user's input to command or instruct the HV module 106 to generate the correct timing or length of time and the correct amplitude and profile of the pulse width wave. A static electroporation configuration may set the meter output 122 to interface with a single PA 124 via a male / female "T-slot," in which only single or multiple cells in a cell suspension from a fluid sample (multi-well) in the chamber of the PA 124 may be electroporated for a certain period of time, depending on the concentration of cells in the cell suspension within the volume of the fluid sample. A flow electroporation configuration can configure the instrument output 122 to interface with a single PA 124 via a pair of diagonally opposed male / female banana plugs, where multiple fluid samples or wells containing cells in cell suspension within the PA 124 can be electroporated automatically and / or autonomously, such that the processor 103 and / or SBC 102 can instruct the HV module 106 to execute a protocol with one or more bursts of several microseconds, each separated by a pause time for the transfer of fluid for the next sample of fluid inside the PA 124. The protocol with one or more microsecond bursts separated by a pause time can be a pulse-width wave. Flow electroporation can be a faster, more efficient method of transfecting large quantities of cells in cell suspension in a fluid equivalent to 1 L within 30 minutes.

[0022] A user may connect a metering device to the calibration port 126 to have the processor 103 and / or SBC 102 simulate a desired pulse-width wave from the metering device, where the HV board 112 may instruct the HV power supply 116 to charge at least one capacitor bank 118 to generate a desired voltage amount for the PM board 120 to generate a pulse-width wave at the meter output 122 via the HV switch 119 based on the pulse-width wave from the user's metering device, but there may not be a PA 124 interfacing with the metering output 122. The calibration port 126 may be used to calibrate the system 100 once or periodically before operation of the electroporation device. In one embodiment, the processor 103 and / or SBC 102 may be connected to the PM board 120 via the SB 104. In another embodiment, the processor 103 and / or SBC 102 may be connected to the PM board 120 via a connection of the SB 104 to the HV board 112. In yet another embodiment, the processor 103 and / or the SBC 102 may be connected to the PM board 120 via a connection of the SB 104 to the HV board 112 and the HV switch 119. In another embodiment, the processor 103 and / or the SBC 102 may be connected to the HV board 112, the HV power supply 116, at least one capacitor bank 118, the HV switch 119, the PM board 120, the meter output 122, the PA 124, and the calibration port 126 via a connection of the SB 104 to the HV module 106.

[0023] The LV module 108 may include an isolated LV DC power supply 144, a display and touch screen 146 or user interface, a barcode reader 148, one or more fans 150 (referred to herein as fans 150), one or more speakers 152 (referred to herein as speakers 152), a power button 154, one or more Universal Serial Bus 3.5 ports 156 (referred to herein as USB 3.0 ports 156), a light emitting diode (LED) board 158 (referred to herein as LED board 158), an inlet pump 160, an inlet valve 162, an outlet pump 164, and an outlet valve 166, all of which may constitute components of the LV module 108.

[0024] SB 104 may gate isolated LV DC power supply 114 from PEM 110 using one or more DC relays (not shown in FIG. 1 ). In another embodiment, isolated LV DC power supply 114 may gate PEM 110 using one or more DC relays (not shown in FIG. 1 ) from LV module 108. In one embodiment, processor 103 and / or SBC 102 may manage, control, and / or collect data from isolated LV DC power supply 144, display and touch screen 146, barcode reader 148, fan 150, speaker 152, power button 154, USB 3.0 port 156, LED board 158, inlet pump 160, inlet valve 162, outlet pump 164, and outlet valve 166 via SB 104 connections. In another embodiment, the SB 104 may be connected to all components within the LV module 108, and the processor 103 and / or SBC 102 may manage and / or control the isolated LV DC power supply 144, the display and touch screen 146, the barcode reader 148, the fan 150, the speaker 152, the power button 154, the USB 3.0 port 156, the LED board 158, the inlet pump 160, the inlet valve 162, the outlet pump 164, and the outlet valve 166.

[0025] In the exemplary embodiment, a display and touchscreen 146 is used. However, other embodiments may alternatively implement other known user interface devices. For example, the display and touchscreen 149 may instead be implemented using a non-touchscreen display and alternative means of user input, such as a keyboard, mouse, programmable softkeys, hard-coded buttons, etc.

[0026] The processor 103 and / or SBC 102 may collect transfection data related to cells in the cell suspension from the fluid sample inside the chamber of the PA 124 for storage in one or more storage devices (not shown in FIG. 1 ) in the system 100. The processor 103 and / or SBC 102 may enable an operator or user to manipulate the data on the display and touch screen 146. The SBC 102 may provide the operator or user with computer functionality in the system 100, where a standalone desktop or laptop computer may not be necessary. The SBC 102 may command, manage, and / or regulate the display and touch screen 146, barcode reader 148, fan 150, speaker 152, power button 154, USB 3.0 port 156, LED board 158, inlet pump 160, inlet valve 162, outlet pump 164, and outlet valve 166. An operator or user may scan the PA 124 containing the fluid sample into the barcode reader 148 in order for the processor 103 and / or SBC 102 to identify and associate a required pulse-width wave profile to apply to cells in a cell suspension from the fluid sample based on a protocol selected by the operator or user. The scanned information, which may be read by the barcode reader 148, may be stored in one or more storage devices for later identification and association with results in the SBC 102 and / or operational data generated by the operator or user at the display and touchscreen 146. The processor 103 and / or SBC 102 may receive one or more commands from the operator or user via the display and touchscreen 146 to execute a particular required pulse-width wave profile for application via the instrument output 122 to cells in a cell suspension from the fluid sample.

[0027] The processor 103 and / or SBC 102 may operate a fan 150 to cool components within the LV module 108. The processor 103 and / or SBC 102 may use a speaker 152 to communicate results to an operator or user based on input from the display and touchscreen 146. The processor 103 and / or SBC 102 may detect via the SB 104 that an operator or user may have pressed a power button 154 so that the processor 103 and / or SBC 102 may power on the system 100. The operator or user may store saved results from the SBC 102 and operational data generated on the display and touchscreen 146 in one or more storage devices for transfer to an external storage device via the USB 3.0 port 156. The power button 154 may be illuminated in one or more colors, such as blue, yellow, red, green, or orange, via the processor 103 and / or SBC 102 using the LED board 158, where a first color may indicate that the system 100 is “on” and a second color may indicate that the system 100 is “off.” Additionally, the processor 103 and / or SBC 102 may use the LED board 158 to illuminate a hollow rectangular portion around the meter output 122 in one or more colors, such as blue, yellow, red, green, or orange. In another embodiment, the processor 103 and / or SBC 102 may illuminate a hollow rectangular portion around the meter output 122 in one or more colors, such as blue, yellow, red, green, or orange. The color of the hollow rectangular portion around the meter output 122 may indicate the progress or status of the electroporation process to an operator or user. Additionally, the processor 103 and / or SBC 102 may change the color of the hollow rectangular shaped portion around the meter output 122 .

[0028] The processor 103 and / or SBC 102 may determine the type of fluid and the viscosity of the fluid within the PA 124 chamber, where a high number of cells in the cell suspension may indicate a high level of viscosity in the fluid, which may require high pressure to move the fluid within the chamber of the PA 124 during flow electroporation. The processor 103 and / or SBC 102 may directly use and / or command the use of the inlet pump 160 and the outlet pump 164. The inlet pump 160 and the outlet pump 164 may be used by circulating air around the chamber of the PA 124 to create positive or negative air pressure to move or draw a fluid sample into or out of the chamber of the PA 124. For example, the inlet pump 160 may be used to create negative or positive air pressure by drawing air out of the chamber within the PA 124 or vice versa, which may draw a fluid sample from a filled 1 L bag into the chamber within the PA 124. After the electroporation process is performed, the outlet pump 164 can generate positive or negative air pressure by blowing air out of the chamber in the PA124 or vice versa, which can draw the fluid sample from the chamber in the PA124 into an empty or filled 1 L bag containing the transfected cells in cell suspension in the fluid sample.

[0029] Processor 103 and / or SBC 102 may use inlet valve 162 or outlet valve 166. Processor 103 and / or SBC 102 may precisely control the amount of fluid circulating from the filled 1 L bag to a chamber in PA 124 and out to the empty or filling 1 L bag based on processor 103 and / or SBC 102 identifying the type and viscosity of the fluid. For example, according to inlet pump 160 and outlet pump 164, processor 103 and / or SBC 102 may activate inlet valve 162 and outlet valve 166 to stop fluid from flowing from the filled 1 L bag to a chamber in PA 124 and out to the empty or filling 1 L bag, even when inlet pump 160 and / or outlet pump 164 may be exerting large positive and / or negative pressures because inlet valve 162 and outlet valve 166 may compress one or more tubes coupled to PA 124 to prevent fluid movement. The processor 103 and / or SBC 102 may accelerate or decelerate the flow of fluid from the filled 1 L bag to the chamber of the PA 124 and into the empty or filling 1 L bag even though the inlet pump 160 and / or outlet pump 164 may be exerting low levels of positive and / or negative pressure, as the inlet valve 162 and outlet valve 166 may compress or release one or more tubes coupled to the PA 124 to slow or accelerate the movement of fluid.

[0030] An operator or user may momentarily press emergency stop (E-stop) button 168 (referred to herein as E-stop button) to immediately stop the electroporation process or the pulse width wave that may be generated by PM board 120 and / or executed by HV board 112 via HV switch 119, where a D flip-flop (not shown in FIG. 1 ) in SB 104 may override, disable, block, and / or bypass processor 103 and / or SBC 102 and directly stop or terminate HV board 112, HV switch 119, and PM board 120 in HV module 106. The D flip-flop may override, disable, block, and / or bypass processor 103 and / or SBC 102 to manage and / or control HV board 106 and its components, and the D flip-flop may still utilize processor 103 and / or SBC 102 to manage and / or control LV module 108 and its components. Additionally, a D flip-flop within the SB 104 may communicate with the processor 103 and / or SBC 102 via the SBC 102's USB 3.0 port to display on the display and touchscreen 146 a status that the E-stop button 168 may have been pressed by an operator or user. The E-stop 168 may override, disable, block, and / or bypass the processor 103 and / or SBC 102 for immediate termination of any electroporation in the HV module 106. Additionally, the E-stop 168 may be implemented to stop the high voltage during the electroporation process for emergency situations where near-instant deactivation may be advantageous. For example, the E-stop 168 may be used to prevent destruction of cells in a cell suspension from a fluid sample, to prevent the system 100 from being destroyed, to prevent any leaking fluid from electrocuting an operator or user, or to prevent an explosion of a flammable fluid sample.Additionally, the E-stop 168 and the D flip-flop in the SB 104 may not rely on software on the system 100 to perform immediate termination of the electroporation process in the HV module 106 when overriding, disabling, blocking, and / or bypassing the processor 103 and / or SBC 102. The E-stop button 168 may be illuminated with a red LED light by the processor 103 and / or SBC 102.

[0031] 2 is a schematic block diagram illustrating an exemplary non-software-based emergency stop function consistent with disclosed embodiments. System 200 may include an E-stop button 202 (see also E-stop button 168 in FIG. 1 ) (E-stop button 168 in FIG. 1 may be the same as E-stop button 202 in FIG. 2 ), an SB 204 (see also SB 104 in FIG. 1 ), a D flip-flop 206, a gate 208, a pulse-forming circuit 210, a PM board 212 (see also PM board 120 in FIG. 1 ), an HV switch 214 (see also HV switch 119 in FIG. 1 ), an SBC 216 (see also SBC 102 in FIG. 1 ), and a display and touchscreen 218 (see also display and touchscreen 146 in FIG. 1 ), all of which may constitute components of system 200. System 200 may be a subcomponent of system 100 in FIG. 1 . The SB 204, the D flip-flop 206, the gate 208, and / or the pulse forming circuit 210 may be electronic circuits.

[0032] The E-stop button 202 is connected to a D flip-flop 206. The D flip-flop 206 may be located inside the SB 204. The D flip-flop 206 is connected to a preset input pin (see Equation 1).

[0033]

number

[0034] , input pin (D), clock input pin (CLK), and clear input pin (see [Equation 2])

[0035]

number

[0036] , power supply pin (VCC), output pin (Q), and inverted output pin (see [Equation 3])

[0037]

number

[0038] To avoid relying on the clocking function of the D flip-flop 206, the input pin (D) may be connected to ground so that the rising edge of the clock input pin (CLK) can be used to reset the D flip-flop 206, the ground pin (GND) may be connected to ground, the power supply pin (VCC) may be connected to a voltage level (+V) required to operate the D flip-flop 206, and the clear input pin (see Equation 4)

[0039]

number

[0040] can also be tied to a logically high voltage level and thus disabled.

[0041] Furthermore, the E-stop button 202 may be configured such that when a user momentarily presses the E-stop button 202, a momentary signal is applied to the preset input pin (see Equation 5).

[0042]

number

[0043] (the instantaneous signal can be held, stored, and / or maintained by the D flip-flop 206)

[0044]

number

[0045] , where the preset input signal may be a logic "1" value. In other embodiments, the preset input signal may be a low voltage or ground connection. Preset Input Pin (see Equation 7)

[0046]

number

[0047] can react immediately when the preset input signal can be pulled low (assigned a logical value of "0") or pulled high (assigned a logical value of "1"), unlike a conventional input pin (D) (not connected to ground but connected to an input signal) which can be asynchronous and only react on the rising edge of the clock signal on the clock input pin (CLK).

[0048]

number

[0049] may be assigned a logical value of "1", where the system 200 may latch onto the signal received from the E-stop button 202, even if the operator or user may subsequently press the E-stop button 202.

[0050] In some embodiments, it may be advantageous for the system 200 to output a latch signal via two separate outputs, where a first output may be a hardware safety mechanism and a second output (see also logic signal) may be a software safety mechanism. The latch signal for the software safety mechanism (second output) may be output directly to the output pin (Q). The latch signal for the hardware safety mechanism (first output) may be output to the inverted output pin (see Equation 9).

[0051]

number

[0052] In a hardware safety mechanism, system 200 may rely solely on D flip-flop 206 to directly deactivate power in HV module 106 (see FIG. 1 ), where D flip-flop 206 may also override, disable, block, and / or bypass logic signals from processor 103 and / or SBC 216 via SB 204. An advantage of a hardware safety mechanism is that it does not rely on software commands and / or logic signals that may be susceptible to timing or logic errors (software delays) in processor 103 and / or SBC 216 that may prevent immediate deactivation of power (electroporation) in HV module 106, where a resulting and / or equal combination of positive and negative pulse profiles or waves corresponding to the required voltage amount may not be immediately applied to a chamber containing cells in a cell suspension from a fluid sample in PA 124 via the chain of PM board 212, HV switch 214, and instrument output 122 (shown in FIG. 1 ). In a software safety mechanism, system 200 may rely on both processor 103 and / or SBC 216 and D flip-flop 206, where D flip-flop 206 may send a latch signal to processor 103 and / or SBC 216 to display a message to an operator or user via display and touchscreen 218 that the electroporation process may have terminated, or D flip-flop 206 may receive input from an operator or user via processor 103 controlled by SBC 216 from display and touchscreen 218 requesting reactivation of the electroporation process. In one embodiment, D flip-flop 206 may receive input from an operator or user via SBC 216 from display and touchscreen 218 requesting reactivation of the electroporation process.In another embodiment, the D flip-flop 206 may receive input from an operator or user via the display and touchscreen 218 through the processor 103 requesting reactivation of the electroporation process. Although the software safety mechanism may be susceptible to timing or logic errors (software delays) in the processor 103 and / or SBC 216 logic signals for operation of components within the LV module 108 and / or HV module 106, those software delays may not inadvertently affect the D flip-flop 206's direct deactivation, override, disable, block, and / or bypass of power for electroporation in the HV module 106.

[0053] Preset input pin (see [Equation 10])

[0054]

number

[0055] The latched signal from may be output directly to the output pin (Q) as a logic "1" which may be communicated to the SBC board 216, where the processor 103 may cause the display and touch screen 218 to display a message to the operator or user that the E-stop function may be active.

[0056]

number

[0057] The latch signal from the inverted output pin (see Equation 12) can be sent as a logic value of “0” to the HV module 106 and / or PM board 212 and HV switch 214 to stop the execution and / or generation of the pulse width wave at the instrument output 122.

[0058]

number

[0059] , where D flip-flop 206 may directly deactivate, override, disable, block, and / or bypass processor 103 and / or SBC 216 from managing and / or controlling HV module 106 and its components of FIG. 1. In another embodiment, gate 208 receives as a first input a logic "1" from its inverted output pin (see Equation 13)

[0060]

number

[0061] Preset input pin (see Equation 14)

[0062]

number

[0063] 1 , and a second input of “1” from pulse forming circuit 210. A first input of “0” combined with a first input of “1” may cause gate 208 to output a logic value of “0” to PM board 212. The output logic value of “0” from gate 208 to PM board 212 may disable or stop HV switch 214 from applying the generated positive and / or negative pulse profiles or waves from PM board 212 to meter output 122 of FIG. 1 . In yet another embodiment, the output logic value of “0” from gate 208 to PM board 212 may disable or stop PM board 212 from generating positive and / or negative pulse profiles or waves, and HV switch 214 may not apply a voltage to meter output 122.

[0064] The display of a message on the display and touch screen 218 may request the user to reset the latched signal from the E-stop 202 to "0" by pressing a graphical user interface (GUI) icon on the display and touch screen 218. A logical "1" value from the user requesting to reset the E-stop button 202 signal may be sent to the clock input pin (CLK), which in combination with input pin (D), which is always set to a logical "0", may be sent to the preset input pin (see Equation 15).

[0065]

number

[0066] to "0". In one embodiment, a signal from the user requesting to reset the E-stop button 202 may be received by the display and touch screen 218, sent to the SBC 216 and / or processor 103, and sent from the SBC 216 and / or processor 103 to a clock input pin (CLK).

[0067]

number

[0068] is reset to a logic value of "0", the output pin (Q) may have a logic value of "0", which may indicate to the SBC 216 and / or processor 103 that the E-stop button 202 may not be pressed and normal operation may continue as described in FIG. 1. Furthermore, the inverted output pin (see Equation 17) of the D flip-flop 206

[0069]

number

[0070] is the preset input pin (see Equation 18)

[0071]

number

[0072] may have a logical value of "1" in response to being reset to a logical value of "0".

[0073]

number

[0074] may allow the processor 103 and / or SBC 216 to instruct the PM board 212 to generate a pulse width wave that is executed by the HV switch 214, where the D flip-flop 206 can no longer deactivate, override, disable, block, and / or bypass the processor 103 and / or SBC 216, requiring the user to reset the E-stop button 202. In another embodiment, the gate 208 may, in normal operation, connect the inverting output pin (see Equation 20)

[0075]

number

[0076] The D flip-flop 206 may receive a logic value of “1” from a first input from the E-stop button 202 and a logic value of “1” from the pulse forming circuit 210 from a second input, which may cause the gate 208 to output a logic value of “1” to the PM board 212. With the output being a logic value of “1” from the gate 208, the PM board 212 and the HV switch 214 may resume normal operation, with the processor 103 and / or the SBC 216 now instructing the PM board 212 to generate a pulse width wave executed by the HV switch 214, and the D flip-flop 206 now requesting that the user reset the E-stop button 202 so that it can no longer override, disable, block, and / or bypass the processor 103 and / or the SBC 216.

[0077] 3 shows a diagram of a display and touchscreen graphical user interface for resetting the E-stop signal, consistent with disclosed embodiments. GUI 300 on display and touchscreen 218 of FIG. 2 (see also display and touchscreen 146 of FIG. 1) may display a message 302 asking, "Reset E-stop signal?" An operator or user may choose to touch or press GUI icon 304, which, when pressed 306 by the user, may cause E-stop 202 to reset to "0." A logic value of "1" from the user requesting that the E-stop button 202 signal be reset may be sent to the clock input pin (CLK), which, combined with input pin (D), which is always set to a logic value of "0" or ground, may be sent to the preset input pin (see Equation 21).

[0078]

number

[0079] may be reset to "0".

[0080] FIG. 4 is an exemplary diagram of an integrated electroporation system and its components in static and flow electroporation configurations consistent with disclosed embodiments. FIG. 4 includes views 402A and 402B of system 400 (see also systems 100 and 200). In view 402A, system 400 may have a bezel subassembly 404 having a "T" shape with a thick, long horizontal portion and a thin, short vertical portion. Bezel subassembly 404 may include a display and touchscreen 406 (see also display and touchscreen 146 in FIG. 1 and display and touchscreen 218 in FIG. 2) in the center of the thick, long horizontal portion and a meter output 408 (see also meter output 122 in FIG. 1) in the center of the thin, short vertical portion. Meter output 408 is surrounded by a hollow rectangular portion 410, which may be illuminated by LED board 158 from FIG. 1. The meter output 408 in view 402A may be configured for static electroporation, where a female "T-slot" 412 of the meter output 408 may interface with a male "T-slot" (not shown in FIG. 4) in the PA 124 of FIG. 1. The female "T-slot" 412 may be illuminated to indicate to an operator or user that the static electroporation configuration may be active. A knob 414 may be recessed beneath the meter output 408 and a thin, short vertical portion of the bezel assembly 404. The knob 414 may be positioned by the operator or user by rotating the knob 414 clockwise to engage the static electroporation configuration. In view 402B, the meter output 408 may have the knob 408 rotated counterclockwise by an operator or user to engage a flow electroporation configuration, where a pair of diagonally opposed female banana plugs 418 of the meter output 408 may interface with a pair of diagonally opposed male banana plugs (not shown in FIG. 4) on the PA 124 of FIG. 1. The pair of diagonally opposed female banana plugs 418 may be illuminated to indicate to the operator or user that the flow electroporation configuration may be active.

[0081] In view 402B of FIG. 4, the system 400 may have a lower fascia subassembly 420 that interfaces with the lower peripheral edge or profile of the bezel subassembly 404. The lower fascia subassembly 420 may include, from view 402A, a barcode reader 422 (see also barcode reader 148 in FIG. 1), an E-stop button 424 (see also E-stop button 168 in FIG. 1 and E-stop button 202 in FIG. 2), a power button 426 (see also power button 154 in FIG. 1), a USB 3.0 port 428 (see also USB 3.0 port 156 in FIG. 1), an inlet pump 430 (see also inlet pump 160 in FIG. 1) having a knob 432 for attaching the tubing of the PA 124 in a flow electroporation configuration, an inlet valve 434 (see also inlet valve 162 in FIG. 1), an outlet pump 436 (see also outlet pump 164 in FIG. 1) having a knob 438 for attaching the tubing of the PA 124 in a flow electroporation configuration, and an outlet valve 440 (see also outlet valve 166 in FIG. 1).

[0082] In view 402A of Figure 4, system 400 may have a left panel 442 that interfaces the left edge or profile of the thickened portion of bezel subassembly 404 with lower fascia subassembly 420. Left panel 442 may include a left hook subassembly 444 in the stowed configuration. In view 402B of Figure 4, system 400 may have a right panel 446 that interfaces the right edge of the thickened portion of bezel subassembly 404 with lower fascia subassembly 420. Right panel 446 may include a right side hook subassembly 448 in the stowed configuration.

[0083] The E-stop button 424 in view 402A may be located in the upper right corner of the lower fascia subassembly 420 in view 402B, in line with the female “T-slot” 412. The E-stop button 424 may be located in a similar location, such as an equivalent location in the left corner of the lower fascia subassembly 420, in other embodiments. Placing the E-stop button 424 in this or a similar location may have certain advantages. For example, the placement of the E-stop button 424 may allow easy access without interfering with one or more tubes of the PA 124, away from potential leaks from the tubes and / or PA 124. This placement may also position or locate the E-stop button 424 closer to the eye level of an operator or user, making it easier to identify the E-stop button 424 in an emergency.

[0084] FIG. 5 is an exemplary diagram of a processing assembly interfacing with a meter output in a static electroporation configuration, consistent with disclosed embodiments. FIG. 5 shows a close-up view of system 500 (see also system 100 of FIG. 1 , system 200 of FIG. 2 , and system 400 of FIG. 4 ), in which PA 502 (see also PA 124 of FIG. 1 ) may be configured for static electroporation. PA 502 may interface with meter output 504 (see also meter output 122 of FIG. 1 and meter output 408 of FIG. 4 ), in which female “T-slot” 412 in FIG. 4 may interface with male “T-slot” (not shown in FIG. 5 ) of meter output 504. A hollow rectangular-shaped portion 506 (see also hollow rectangular-shaped portion 410 of FIG. 4 ) around meter output 504 may be illuminated by LED board 158 from FIG. 1 to indicate to an operator or user that system 500 may be in a static electroporation configuration.

[0085] Figure 6 is an exemplary diagram of a processing assembly interfacing with a meter output in a flow electroporation configuration, consistent with disclosed embodiments. Figure 6 illustrates a system 600 (see also system 100 of Figure 1, system 200 of Figure 2, system 400 of Figure 4, and system 500 of Figure 5) using views 602A, 602B, and 602C. View 602A of Figure 6 includes a bezel subassembly 604 (see also bezel subassembly 404 of Figure 4) having a display and touchscreen 606 (see also display and touchscreen 146 of Figure 1, display and touchscreen 218 of Figure 2, and display and touchscreen 406 of Figure 4) and a hollow rectangular-shaped portion 608 (see also hollow rectangular-shaped portion 410 of Figure 4 and hollow rectangular-shaped portion 506 of Figure 5) around a meter output 504 (not shown in Figure 6).

[0086] View 602A in FIG. 6 shows a lower fascia subassembly 610 (see also lower fascia subassembly 420 in FIG. 4) that includes an inlet pump 612 (see also inlet pump 160 in FIG. 1 and inlet pump 430 in FIG. 4), an inlet valve 614 (see also inlet valve 162 in FIG. 1 and inlet valve 434 in FIG. 4), an outlet pump 616 (see also outlet pump 164 in FIG. 1 and outlet pump 436 in FIG. 4), and an outlet valve 618 (see also outlet valve 166 in FIG. 1 and outlet valve 440 in FIG. 4).

[0087] In one embodiment, a PA configured for flow electroporation may include a filled 1 L bag 620, an inlet tube 622, a chamber 624, an inlet / outlet tube 626, an inlet / outlet tube 628, an outlet tube 630, and an empty or filling 1 L bag 632. The filled 1 L bag 620 may be connected to the inlet tube 622, which may be connected to the inlet valve 614. The inlet tube 622 may enter the chamber 624, and the inlet / outlet tube 626 may exit the chamber 624 and be connected to the inlet pump 612. The inlet / outlet tube 626 may exit the inlet pump 612, and an end orifice of the inlet / outlet tube 626 may be exposed to air.

[0088] Additionally, an inlet / outlet tube 628 may exit chamber 624 and be connected to outlet pump 616. Inlet / outlet tube 628 may exit outlet pump 616, and an end orifice of inlet / outlet tube 628 may be exposed to air. Additionally, outlet tube 630 may exit chamber 624 and be connected to outlet valve 618. Outlet tube 630 may exit outlet valve 618 for connection to an empty or filling 1 L bag 632.

[0089] During operation, inlet pump 612 pressurizes inlet / outlet tube 626, chamber 624, and inlet tube 622 to blow air out the end orifice of inlet / outlet tube 626, which may cause fluid from filled 1 L bag 620 to flow through inlet tube 622, regulated by valve 614, and into chamber 624. Similarly, outlet pump 612 pressurizes inlet / outlet tube 628, chamber 624, and outlet tube 630 to blow air out the end orifice of inlet / outlet tube 628, which may cause fluid within chamber 624 to flow through outlet tube 630, regulated by valve 618, and into empty or filling 1 L bag 632. The fluid within chamber 624 may be electroporated by meter output 504. A pair of diagonally opposed male banana plugs (not shown in FIG. 6) from chamber 624 may interface with a pair of diagonally opposed female banana plugs 418 (not shown in FIG. 6) on instrument output 504. SB 204 may manage and regulate fluid flow in chamber 624 by controlling inlet pump 612, inlet valve 614, outlet pump 616, and outlet valve 618. For example, SB 204 may cause inlet pump 612 and outlet pump 616 to draw air through the end orifices of inlet / outlet tubing 626 and the outlet orifice of inlet / outlet tubing 628, respectively, to prevent fluid from flowing in or out of chamber 624 during electroporation, where inlet valve 614 may prevent fluid flow in inlet tubing 622 and outlet valve 618 may prevent fluid flow in outlet tubing 630. SB 204 can determine, via current sensor 134, the appropriate amount of fluid in chamber 624 for electroporation.

[0090] View 602B in Figure 6 shows a left panel 634 (see also left panel 442 in Figure 4) with a left hook subassembly 636 (see also left hook subassembly 444 in Figure 4) in an unfolded configuration, where the left hook subassembly 634 can support a filled 1 L bag 620 in view 602A. View 602C in Figure 6 shows a right panel 638 (see also left panel 446 in Figure 4) with a right hook subassembly 640 (see also left hook subassembly 448 in Figure 4) in an unfolded configuration, where the right hook subassembly 640 can support an empty or filling 1 L bag 632 in view 602A.

[0091] FIG. 7 is an exemplary diagram of an electroporation system with low-voltage component packaging and a display and touchscreen consistent with disclosed embodiments. It shows a subsystem 700 with the LV module 108 of FIG. 1 and the SB 104 of FIG. 1. The subsystem 700 includes a bezel subassembly 702 (see also bezel subassembly 404 of FIG. 4 and bezel subassembly 606 of FIG. 6), a lower fascia subassembly 704 (see also lower fascia subassembly 420 of FIG. 4 and lower fascia subassembly 610 of FIG. 6), a front panel subassembly 706, and a display and touchscreen 708 (see also display and touchscreen 146 of FIG. 1, display and touchscreen 148 of FIG. 2). 4 and 606 of FIG. 6), SBC 710 (see also SBC 102 of FIG. 1 and SBC 212 of FIG. 2), barcode reader 712 (see also barcode reader 148 of FIG. 1 and barcode reader 422 of FIG. 4), and meter output 714 (see also meter output 122 of FIG. 1, meter output 408 of FIG. 4, and meter output 504 of FIG. 5). 1, an LED board 716 (see also LED board 158 in FIG. 1), an outlet pump 718 (see also outlet pump 164 in FIG. 1, outlet pump 436 in FIG. 4, and outlet pump 616 in FIG. 6), an outlet valve 720 (see also outlet valve 166 in FIG. 1, outlet valve 440 in FIG. 4, and outlet valve 618 in FIG. 6), an inlet pump 722 (see also inlet pump 160 in FIG. 1, inlet pump 430 in FIG. 4, and inlet pump 612 in FIG. 6), and an inlet valve 724 (see also inlet pump 160 in FIG. 1, inlet pump 430 in FIG. 4, and inlet pump 612 in FIG. 6). 4 and inlet valve 614 in FIG. 6), a power button 726 (see also power button 154 in FIG. 1 and power button 426 in FIG. 4), a USB 3.0 port 728 (see also USB 3.0 port 156 in FIG. 1 and USB 3.0 port 156 in FIG. 4), and an E-stop 730 (see also E-stop button 168 in FIG. 1, E-stop button 202 in FIG. 2 and E-stop button 424 in FIG. 4).

[0092] As shown in FIG. 7 , a bezel subassembly 702 and a lower fascia subassembly 704 may be attached to the front panel subassembly 706 facing the front panel subassembly. A display and touch screen 708 may be attached to the bezel subassembly 702. An SBC 710 may be attached to the upper left side of the front panel subassembly 706. A barcode reader 712 may be attached to the upper right side of the lower fascia subassembly 704 and may be attached below the SBC 710 on the front panel subassembly 706. A gauge output 714 may be attached to the bezel subassembly 702. An LED board 716 may be attached to the gauge output 714 facing the front panel subassembly 706. An outlet pump 718 may be attached to the lower left side of the lower fascia subassembly 704 and the lower left side of the front panel subassembly 706. An outlet valve 720 may be attached to the right of the outlet pump 718 in the center of the lower fascia subassembly 704. An inlet pump 722 may be mounted to the lower right of the lower fascia subassembly 704 and the lower right of the front panel subassembly 706. An inlet valve 724 may be mounted to the left of the inlet 722 and to the right of the outlet valve 720 in the center of the lower fascia subassembly 704. A power button 726 may be mounted to the lower center of the lower fascia subassembly below the outlet valve 720. A USB 3.0 port 728 may be mounted to the right of the power button 726 in the center of the lower fascia subassembly 704 below the inlet valve 724. An E-stop button 730 may be mounted to the upper left of the fascia subassembly 704.

[0093] 8 shows an exemplary block diagram of an exemplary method for electroporation of cells in a fluid cell suspension, consistent with some embodiments of the present disclosure. Method 800, as shown in FIG. 8, may be performed by a system board that may receive a signal from E-stop button 168 to stop the voltage applied by meter output 122 to the cells in the fluid cell suspension, at block 802. This signal may be a momentary signal generated the moment the button on the other signaling device is activated and may act as an interruption of the electroporation process for the cells in the fluid cell suspension.

[0094] In block 804, the method 800 may be executed by a system board that may latch a signal from the E-stop button 168. The signal may be latched when the input momentary signal is received without waiting for a clock signal. The signal may remain latched even when the input momentary signal is no longer being received.

[0095] In block 806, the method 800 may be performed by a system board that may block logic signals that may control the voltage supply to the meter outputs. At least one processor may send logic signals to control the power supply to the meter outputs.

[0096] In block 808, the method 800 may be performed by a system board that may terminate the voltage applied to the cells in the cell suspension in fluid by the meter output 122. This termination may occur nearly instantaneously without waiting for further input from the processor 103.

[0097] In block 810, the method 800 may be executed by a system board that may disable the processor 103 from providing instructions to the high voltage module 106 to apply a voltage to cells in a cell suspension in a fluid via the meter output 122.

[0098] In block 812, the method 800 may be executed by the processor 103, which may send a message to the user via the display and touchscreen device 146 in the low voltage module 108 to reset the signal from the E-stop 168. This signal may be sent after block 808 is executed, independently of block 810. Blocks 808 and 810 may occur in parallel.

[0099] While the present disclosure has been shown and described with reference to specific embodiments thereof, it will be understood that the present disclosure can be implemented without modification in other environments. The foregoing description has been presented for illustrative purposes. It is not exhaustive and is not limited to the precise form or embodiment disclosed. Modifications and adaptations will be apparent to those skilled in the art from consideration of the specification and practice of the disclosed embodiments. Additionally, while aspects of the disclosed embodiments are described as being stored in memory, those skilled in the art will understand that these aspects can also be stored on other types of computer-readable media, such as a secondary storage device, e.g., a hard disk or CD ROM, or other forms of RAM or ROM, USB media, DVD, Blu-ray, or other optical drive media.

[0100] Computer programs based on the specification and disclosed methods are within the skill of an experienced developer. Various programs or program modules can be created using any of the techniques known to those skilled in the art and can be designed in combination with existing software. For example, program sections or program modules can be designed in or with the .NET Framework, the .NET Compact Framework (and related languages ​​such as Visual Basic, C, etc.), Java, C++, Objective-C, HTML, HTML / AJAX combinations, XML, or HTML, including Java applets.

[0101] Furthermore, while exemplary embodiments have been described herein, a range of embodiments having equivalent elements, modifications, omissions, combinations (e.g., of aspects across various embodiments), adaptations, and / or alterations will be understood by those skilled in the art based on this disclosure. Any limitations in the claims should be interpreted broadly based on the language used in the claims and should not be limited to the examples described herein or during the prosecution of this application. Examples should be construed as non-exclusive. Furthermore, the steps of the disclosed methods may be modified in any manner, including changing the order of steps and / or inserting or deleting steps. Accordingly, it is intended that the specification and examples be considered exemplary only, with the true scope and spirit being indicated by the appended claims and their full scope of equivalents. [Explanation of symbols]

[0102] 100 systems 102 Single Board Computer (SBC), SBC 103 processors 104 System board (SB), SB 106 High Voltage (HV) Module, HV Module 108 Low Voltage (LV) Module, LV Module 110 Power Entry Module (PEM), PEM 112 High voltage board, HV board 114 Isolated low voltage (LV) DC power supply, LV DC power supply 116 High voltage power supply 116, HV power supply 118 Capacitor Bank 119 High voltage switch 119, HV switch 120 Pulse Modulator (PM) Board, PM Board 122 Instrument Output 124 Processing Assembly (PA), PA 126 Calibration Port 134 Current Sensor 144 Isolated LV DC power supply 146 Displays and Touchscreens 148 Barcode Reader 150 fans 152 Speaker 154 Power button 156 Universal Serial Bus 3.5 ports, USB 3.0 ports 158 Light Emitting Diode (LED) Board, LED Board 160 Inlet pump 162 Inlet valve 164 Outlet Pump 166 Outlet Valve 168 Emergency stop (E-stop) button, E-stop button, E-stop 200 systems 202 E-stop button 204SB 206 D Flip-Flop Gate 208 210 Pulse forming circuit 212 PM Board 214 HV switch 216 SBC 218 Displays and Touchscreens 300 GUI 302 Message 304 GUI icons 400 System 402A View 402B Views 404 Bezel Sub-Assembly 406 Display and Touchscreen 408 Instrument Output 410 Hollow rectangular part 412 Scalpel "T-slot" 414 Nobu 416 Nobu 418 female banana plug 420 Lower Fascia Sub-Assembly 422 Barcode Reader 424 E-stop button 426 Power Button 428 USB 3.0 ports 430 Inlet Pump 432 Nobu 434 Inlet Valve 436 Outlet Pump 438 Nobu 440 Outlet Valve 442 Left Panel 444 Left Hook Sub-Assembly 446 Right Panel 448 Right Side Hook Sub-Assembly 500 Systems 502 PA 504 Instrument Output 600 System 602A View 602B Views 602C View 604 Bezel Sub-Assembly 606 Display and Touchscreen 608 Hollow rectangular part 610 Lower Fascia Sub-Assembly 612 Inlet pump 614 Inlet valve 616 Outlet Pump 618 Outlet Valve 620 filled 1L bags 622 Inlet tube 624 Chamber 626 Inlet / Outlet Tube 628 Inlet / Outlet Tube 630 Outlet Tube 632 empty or filled 1L bags 634 Left side panel 636 Left Hook Sub-Assembly 638 Right Panel 640 Right Hook Sub-Assembly 700 Subsystem 702 Bezel Sub-Assembly 704 Lower Fascia Sub-Assembly, Fascia Sub-Assembly 706 Front Panel Subassembly 708 Display and Touchscreen 710 SBC 712 Barcode reader 714 Instrument Output 716 LED board 718 Outlet Pump 720 Outlet Valve 722 Inlet Pump 724 Inlet Valve 726 Power Button 728 USB 3.0 ports 730 E-stop

Claims

1. 1. An apparatus for electroporation of cells in a cell suspension in a fluid, said apparatus comprising: a memory for storing instructions; a high voltage module including an instrument output configured to apply a voltage to cells in a cell suspension in a fluid; a low voltage module including a user interface device; a system board connected to the high voltage module, the low voltage module, and a single board computer, the single-board computer includes at least one processor configured to (i) send instructions to control the high voltage module and the low voltage module, and (ii) receive instructions from the user interface device to adjust the voltage applied to the cells in the cell suspension in the fluid by the meter output; and an emergency stop button coupled to the system board; Equipped with The system board receiving a signal from the emergency stop button to stop the voltage applied to the cells in the cell suspension in the fluid by the meter output; latching the signal from the emergency stop button; blocking a logic signal that controls the voltage supply to the meter output; terminating the voltage applied to the cells in suspension in the fluid by the meter output; disabling the at least one processor from providing commands to the high voltage module; further comprising electronic circuitry configured to execute instructions for performing steps including: Device.

2. 10. The apparatus of claim 1, wherein the high voltage module includes a high voltage board and a pulse modulator board configured to implement a positive pulse wave at the meter output.

3. 3. The apparatus of claim 2, wherein the high voltage board and the pulse modulator board are configured to generate a positive pulse wave using at least one bank of capacitors.

4. 10. The apparatus of claim 1, wherein the high voltage module includes a high voltage board and a pulse modulator board configured to implement a negative pulse wave at the meter output.

5. 5. The apparatus of claim 4, wherein the high voltage board and the pulse modulator board generate the negative pulse wave using at least one bank of capacitors.

6. 10. The apparatus of claim 1, wherein a single-board computer is configured to process electroporation results of the cells in the cell suspension in the fluid.

7. The apparatus of claim 1 , wherein the single-board computer is configured to manage the user interface.

8. 10. The apparatus of claim 1, wherein the high voltage module includes a calibration port configured to simulate positive and negative pulse waves at the meter output.

9. 2. The device of claim 1, wherein the electronic circuit includes a D flip-flop that connects to the emergency stop button via a preset pin and terminates the voltage applied to the cells in the cell suspension in the fluid by the meter output via an inverted output pin.

10. 10. The apparatus of claim 1, wherein the electronic circuit includes a D flip-flop connected to the processor via an output pin, and the at least one processor connects to the user interface device via a clock input pin for resetting the signal from the emergency stop button.

11. 1. A method for performing a safety step to terminate electroporation of cells in a cell suspension in a fluid, said method comprising: receiving a signal from the emergency stop button to stop the voltage applied by the meter output to the cells in the cell suspension in the fluid; latching the signal from the emergency stop button; blocking a logic signal that controls the voltage supply to the meter output; terminating the voltage applied to cells in the cell suspension in the fluid by the meter output; disabling the at least one processor from providing commands to a high voltage module; A method comprising:

12. 12. The method of claim 11, wherein the high voltage module includes a high voltage board and a pulse modulator board for implementing a positive pulse wave at the meter output.

13. The method of claim 12 , wherein the high voltage board and the pulse modulator board generate the positive pulse wave using at least one bank of capacitors.

14. 12. The method of claim 11, wherein the high voltage module includes a high voltage board and a pulse modulator board for implementing a negative pulse wave at the meter output.

15. 15. The method of claim 14, wherein the high voltage board and the pulse modulator board use at least one bank of capacitors to generate the negative pulse wave.

16. The method of claim 11 , wherein the single-board computer is configured to manage the user interface.

17. The method of claim 11 , wherein the high voltage module includes a calibration port configured to simulate positive and negative pulse waves at the meter output.

18. 12. The method of claim 11, wherein the electronic circuit includes a D flip-flop connecting to the at least one processor via an output pin, and the at least one processor connecting to a user interface device via a clock input pin for resetting the signal from the emergency stop button.

19. 12. The method of claim 11, wherein the electronic circuit includes a D flip-flop that uses an output pin connected to a single-board computer that manages a user interface, a clock input pin connected to the single-board computer that manages the user interface, and an input pin for resetting the signal from the emergency stop button.

20. 1. An apparatus for electroporation of cells in a cell suspension in a fluid, said apparatus comprising: a memory for storing instructions; a high voltage module including an instrument output configured to apply a voltage to cells in a cell suspension in a fluid; a low voltage module including a user interface device; a system board connected to the high voltage module, the low voltage module, and a single board computer, the single-board computer includes at least one processor configured to (i) send instructions to control the high voltage module and the low voltage module, and (ii) receive instructions from the user interface device to adjust the voltage applied to the cells in the cell suspension in the fluid by the meter output; and an emergency stop button coupled to the system board; a D flip-flop in the system board, the D flip-flop having a preset input pin connected to the emergency stop button, an output pin and a clock input pin connected to the low voltage module, and an inverted output pin connected to the high voltage module; Equipped with The D flip-flop is receiving a signal from the emergency stop button to stop the voltage applied by the meter output to cells in the cell suspension of the fluid; latching the signal from the emergency stop button; blocking a logic signal that controls the voltage supply to the meter output; terminating the voltage applied to the cells in suspension in the fluid by the meter output; disabling the at least one processor from providing commands to the high voltage module; configured to execute instructions to perform steps including: Device.

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