Apparatus and method for electroporation

Abstract

A device and method for electroporation of cells is disclosed. The device includes a memory storing instructions, a high voltage module applying a voltage to the cells, a low voltage module including a user interface, a system board connected to the high voltage module, the low voltage module, and a single board computer, and the single board computer including at least one processor to control the high voltage module and the low voltage module. The system board includes an emergency stop button and an electronic circuit. The electronic circuit receives a signal from the emergency stop button, latches the signal from the emergency stop button, blocks a logic signal that controls the voltage delivered to the instrument output, terminates the voltage applied to the cell, and disables the at least one processor from providing a command to the high voltage module.

Description

[0001] Priority

[0002] This application claims priority to U.S. Provisional Application No. 63 / 404,157, filed on September 6, 2022, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure generally relates to devices, configurations, and methods for electroporation. In particular, embodiments of the present disclosure relate to creative and unconventional devices and methods for providing electroporation of cells in a cell suspension from a fluid sample, where the device integrates a display and touchscreen, a computer, applying a high voltage to cells in a cell suspension from a fluid sample, and a non-software-based emergency stop feature. Background Art

[0004] An electroporation system can apply a voltage to cells in a cell suspension in a fluid through an electric field to increase the permeability of the cell membrane, thereby allowing chemicals, drugs, or nucleic acids (DNA or RNA) to be introduced into the cells. The application of the voltage can cause the electric field to follow a pulse duration schedule or experimental protocol. Electroporation systems can be used in medical, testing, and microbiology applications.

[0005] Current electroporation systems can be stand-alone systems that are only designed for transfection, which may require purchasing a separate desktop or laptop computer to process the results from the current electroporation system. For example, an operator or user of a current electroporation system may need to additionally purchase a desktop or laptop computer to analyze and generate plots from the current electroporation system, where the current electroporation system may only provide raw data to the operator or user. A non-integrated electroporation system that requires purchasing a separate desktop or laptop computer may additionally have problems with compatibility between the operating system of the desktop or laptop computer and the electroporation system. For example, a current electroporation system can provide separate software to an operator or user to install on a desktop or laptop computer to process the raw data generated by the current electroporation system, where the software can help generate plots, identify, and analyze transfected cells in a cell suspension in a fluid. In other cases, the software provided for installation on a desktop or laptop computer may be incompatible with or not up-to-date with the existing operating system, which may force the operator or user to maintain an older version of the operating system of the desktop or laptop computer. In addition, current electroporation systems may need to use a separate storage medium to receive the results of the transfection process in the form of a file containing the raw data, and this separate storage medium can in turn be used on a desktop or laptop computer for the file to process the results. Using a separate storage medium may also require using software to process the results in the file.

[0006] The non-integration of computer functionality in current electroporation systems significantly increases the cost to the operator or user, as they may be forced to purchase a separate electroporation system and a desktop or laptop computer, and incur costs due to converting the current operating system on the desktop or laptop computer to an older operating system compatible with the software provided by the electroporation system. Additionally, the non-integration of computer functionality may also increase the difficulty of operating those systems, as the operator or user 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 may not provide the status of the transfection process. For instance, manufacturers of current electroporation systems may rely on separate software installed on a desktop or laptop computer to provide the electroporation status to the operator or user; however, due to the non-integration between the electroporation system, the desktop or laptop computer, and the software, these statuses may be delayed.

[0007] Furthermore, current electroporation systems that are non-integrated with a desktop or laptop computer may not provide safety features that allow the operator or user to immediately terminate the transfection process in hazardous situations where the operator or user may discover a malfunctioning electroporation system, a fluid sample prepared for processing that is incorrect, a combustible fluid sample, or a leak of conductive fluid samples either inside or outside the electroporation system (which may pose a critical situation to the operator or user). For example, the operator or user may be forced to unplug the power cord of the electroporation system or disconnect the desktop or laptop computer from the electroporation system together, which may also create a highly dangerous situation due to the risk of electric shock or fire. Non-integrated electroporation systems may allow the operator or user to use a command on the desktop or laptop computer (such as the space bar of the keyboard) to terminate the electroporation process; however, this implementation using a desktop or laptop computer with a non-integrated electroporation system may not be instantaneous, as the operator may have to navigate through a series of menus before the software recognizes that the use of the space bar on the keyboard may mean terminating the electroporation process. Additionally, the space bar application for terminating the electroporation process can be software-dependent, where the software may be limited by errors, delays, and the steps of the logical sequence, which may increase the time taken to terminate the operation of the electroporation system. Moreover, the software-based space bar application for terminating the electroporation process may be vulnerable to communication errors between the desktop or laptop computer and the current electroporation system.

[0008] The lack of means to immediately terminate the transfection process increases the cost to the operator and user, as the electroporation system is damaged, the desktop or laptop computer is damaged, the fluid sample is lost, or serious harm is caused to the operator and user.

[0009] Accordingly, there is a need for improved devices and methods for fluid electroporation that integrate computer functionality, apply higher field strengths in volts per meter, apply to higher volumes of fluid up to 1 liter (1L), apply to larger culture sizes, and incorporate a non-software-based emergency stop feature to immediately terminate the electroporation process to protect the fluid sample, the electroporation device, and the operator and user. Summary of the Invention

[0010] One aspect of the present disclosure relates to a device for electroporating cells in a cell suspension in a fluid. The device may include: a memory 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 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 by the instrument output to cells in a cell suspension in a fluid; and an emergency stop button connected to the system board. The system board may further include an electronic circuit configured to run instructions to perform steps including: receiving a signal from the emergency stop button to stop the voltage applied by the instrument output to cells in a cell suspension in a fluid, latching the signal from the emergency stop button, blocking a logic signal controlling voltage delivery to the instrument output, terminating the voltage applied by the instrument output to cells in the suspension in the fluid; and disabling the at least one processor from providing instructions to the high voltage module.

[0011] Another aspect of the present disclosure relates to a method of performing safety steps for terminating the electroporation of cells in a cell suspension in a fluid. The method may include the steps of: receiving a signal from the emergency stop button to stop the voltage applied by the instrument output to cells in a cell suspension in a fluid, latching the signal from the emergency stop button, blocking a logic signal controlling voltage delivery to the instrument output, terminating the voltage applied by the instrument output to cells in a cell suspension in a fluid, and disabling the at least one processor from providing commands to the high voltage module.

[0012] Another aspect of the present disclosure relates to an apparatus for electroporating cells in a cell suspension in a fluid. The apparatus may include: a memory 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 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 by the instrument output to cells in a cell suspension in a fluid; an emergency stop button connectable to the system board; and a D flip - flop in the system board, the D flip - flop may have a preset input pin connected to the emergency stop button, an output pin connected to the low voltage module, a clock input pin, and an inverted output pin connected to the high voltage module. The D flip - flop in the system board may be configured to run instructions to perform steps including: receiving a signal from the emergency stop button to stop the voltage applied by the instrument output to cells in a cell suspension in a fluid; latching the signal from the emergency stop button; blocking the logic signal controlling the voltage delivery to the instrument output; terminating the voltage applied by the instrument output to cells in a cell suspension in a fluid; and disabling the at least one processor from providing commands to the high voltage module.

[0013] Other systems, devices, and methods are also discussed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 is a schematic block diagram showing an exemplary embodiment of an integrated electroporation device with a non - software - based emergency stop consistent with the disclosed embodiments.

[0015] Figure 2 is a schematic block diagram showing an exemplary non - software - based emergency stop feature consistent with the disclosed embodiments.

[0016] Figure 3 depicts an illustration of a graphical user interface of a display and touch screen for resetting an emergency stop signal consistent with the disclosed embodiments.

[0017] Figure 4 is an exemplary illustration of an integrated electroporation system and its components in static and flow - through electroporation configurations consistent with the disclosed embodiments.

[0018] Figure 5 is an exemplary illustration of a processing component interfacing with an instrument output in a static electroporation configuration consistent with the disclosed embodiments.

[0019] Figure 6An exemplary illustration of a processing component interfacing with an instrument output in a flow-through electroporation configuration, consistent with the disclosed embodiments.

[0020] Figure 7 An exemplary illustration of an electroporation system with a low-voltage component enclosure, a display, and a touchscreen, consistent with the disclosed embodiments.

[0021] Figure 8 An exemplary block diagram showing an exemplary method for electroporating cells in a cell suspension in a fluid, consistent with some embodiments of the present disclosure. Detailed Description

[0022] The following detailed description refers to the accompanying drawings. Whenever possible, the same reference numerals are used in the drawings and the following description to refer to the same or like parts. Although several illustrative embodiments are described herein, modifications, adaptations, and other implementations are possible. For example, components and steps shown in the drawings may be replaced, added, or modified, and the illustrative methods described herein may be modified by replacing, reordering, removing, or adding steps to the disclosed methods. Accordingly, the following detailed description is not limited to the disclosed embodiments and examples. Instead, the proper scope of the present invention is defined by the appended claims.

[0023] Embodiments of the present disclosure relate to devices and methods for electroporation that are 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 to introduce chemical or biological samples. Additionally, embodiments of the present disclosure relate to devices and methods for electroporation that are configured to transfect cells in a cell suspension for a fluid volume, such as 1 L, of a culture size containing, for example, 200 billion cells. Further, embodiments of the present disclosure relate to devices and methods for electroporation that are configured to integrate computer functionality including a display and a touchscreen without the need to utilize a separate desktop or laptop computer for ease of use. Moreover, embodiments of the present disclosure relate to devices and methods for electroporation that are configured to include an emergency stop feature or a safety stop feature that is non-software-based and provides immediate termination of the electroporation process to reduce cell damage, device damage, and safeguard the safety of the operator or user.

[0024] Figure 1 A schematic block diagram showing an exemplary embodiment of an integrated electroporation device with a non-software-based emergency stop, consistent with the disclosed embodiments. As Figure 1 shown, system 100 may include one or more devices - components that may constitute system 100. System 100 may include one or more memory storage devices (Figure 1 not shown herein), a single-board computer (SBC) 102 (referred to herein as SBC 102) that may include 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). The processor 103 and / or the SBC 102 may control, manage, and / or collect data in the system 100. The processor 103 may be an ASIC (application-specific integrated circuit) or it may be a general-purpose processor. The processor 103 may include more than one processor. For example, the processors may be positioned in parallel, in series, or in parallel and in series to process all or part of the computer instructions to be processed. In one embodiment, the processor 103 may be in a dedicated circuit board independent of the SBC 102 and / or inside 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 SBC 102, 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) through the connections of the HV module 106 and the LV module 108 to 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 the system 100) may generate a 24-volt (24V) direct current (DC) voltage rail to power the SBC 102, the processor 103, the SB 104, the LV module 108, and the HV module 106. The SB 104 may connect the PEM 110 to the SBC 102 and its components, the processor 103, the HV module 106 and its components, and the LV module 108 and its components. The processor 103 and / or the SBC 102 may manage and / or control the voltage applied to the components in the HV module 106 and / or the LV module 108. The SB104 may use one or more DC relays ( Figure 1 not shown herein) to gate the 24V DC voltage rail from the PEM 110 to power the components inside the SB 104 and inside the HV module 106. Similarly, the LV module 108 may include one or more DC relays ( Figure 1(not shown in the figure) to gate the 24V DC voltage rail from the PEM 110 to power the components inside the SBC 102, the processor 103, and the LV module 108. In another embodiment, the SB 104 may include one or more DC relays to gate the 24V DC voltage rail from the PEM 110 to power its components and the components in the HV module 106, the LV module 108, the processor 103, and the SBC 102. In yet another embodiment, the PEM 110 may include one or more DC relays to gate the 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. In yet another embodiment, the PEM 110 may be directly connected to one or more DC relays or power supplies to gate the 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.

[0025] The HV module 106 may include a high-voltage board 112 (referred to herein as the HV board 112), an isolated low-voltage (LV) DC power supply 114 (referred to herein as the isolated LV DC power supply 114), a high-voltage power supply 116 (referred to herein as the HV power supply 116), at least one capacitor bank 118 including one or more capacitors, a high-voltage switch 119 (referred to herein as the HV switch 119), a pulse modulator (PM) board 120 (referred to herein as the PM board 120), an instrument output 122, a processing assembly (PA) 124 (referred to herein as the PA 124), and a calibration port 126, all of which may constitute the components of the HV module 106.

[0026] The HV board 112 may use a DC relay ( Figure 1The gating (not shown in the figure) isolates the LV DC power supply 114 from the PEM 110 to supply the HV power supply 116. The PEM 110 can supply power to the isolated LV DC power supply 114 to power the HV power supply 116. The HV board 112 can manage and / or regulate the required amount of power to the HV power supply 116 based on the required power demand directed by the processor 103 and / or the SBC 102. The processor 103 and / or the SBC 102 can be connected to the HV board 112 through the SB 104. The HV power supply 116 can in turn charge at least one capacitor bank 118 to maintain or quickly generate the amount of voltage required for electroporation. The required amount of voltage can be in the range between -1000 volts and +1000 volts, where the required amount of voltage can be + / -600 volts, + / -700 volts, + / -800 volts, + / -900 volts, + / -1000 volts and / or higher. The at least one capacitor bank 118 can include one or more capacitors to generate the required amount of voltage. In one embodiment, the at least one capacitor bank 118 can include 4 capacitors capable of generating the required amount of voltage. In another embodiment, the at least one capacitor bank 118 can include one or more capacitors to produce the required amount of voltage.

[0027] The PM board 120 can generate positive and / or negative pulse profiles or waves, which can be run by the HV switch 119 to the instrument output 122. The HV power supply 116 can draw power from the LV DC power supply 114 to charge at least one capacitor bank 118, thereby applying a voltage to the HV switch 119. The PM board 120 can generate positive and / or negative pulse profiles or waves to the HV switch 119. The positive and / or negative pulse profiles or waves from the SBC 102, the processor 103, the SB 104, and / or the PM board 120 can be based on the cells in the cell suspension of the fluid contained in the chamber of the PA 124 that the SBC 102 may have identified. The PM board 120 can run or apply a pulse-width wave generated by the SBC 102 and / or the processor 103, and the pulse-width wave is equivalent to, results in, and / or is equal to the required amount of voltage applied to the instrument output 122 via the HV switch 119. In another embodiment, the PM board 120 can generate a pulse-width wave. In yet another embodiment, the processor 103 can generate a pulse-width wave. In one embodiment, the PM board 120, the SBC 102, and / or the processor 103 can generate a pulse-width wave. The voltage in the instrument output 122 can be applied to the chamber of the PA 124, which contains the cells in the cell suspension of the fluid sample. The pulse-width wave can be a combination of a positive pulse profile or wave and a negative pulse profile or wave, and the pulse-width wave can be equivalent to, result in, and / or be equal to the required amount of voltage applied to the chamber in the PA 124 via the HV switch 119 and the instrument output 122, and the chamber contains the cell suspension of the fluid sample. In another embodiment, the PM board 120 can generate positive and negative pulse profiles or waves for the HV board 112, the SBC 102, the processor 103, and / or the PM 120 to run an amount of voltage equivalent to, resulting in, and / or equal to the required amount of voltage applied to the chamber in the PA 124 via the HV switch 119 and the instrument output 122, and the chamber contains the cell suspension of the fluid sample.

[0028] The processor 103 and / or the SBC 102 can apply an experimental protocol, which can include timing or time length and the correct amplitude and profile of the pulse-width wave. The processor 103 and / or the SBC 102 can command, manage, and / or regulate the HV board 112, the HV switch 119, the PM board 120, and the instrument output 122. In one embodiment, the processor 103 and / or the SBC 102 can also detect that there may be no fluid in the chamber of the PA 124, or can also detect that there may be no PA component inserted into the instrument output 122.

[0029] The instrument output 122 can be configured to provide two operating modes: static electroporation and flow-through electroporation. The instrument output 122 can have a knob or other device that an operator or user can use to set whether the system 100 can operate as (on a small scale) static electroporation or (on a large scale) flow-through electroporation. The processor 103 and / or the SBC 102 can detect the position of the knob or other device on the instrument output 122 based on the user's input to command or instruct the HV module 106 to generate an experimental protocol with the correct timing or time length and the correct amplitude and profile for the pulse width wave. The static electroporation configuration can set the instrument output 122 to interface with a single PA 124 through a male / female "T-slot", where only a single cell or multiple cells in the cell suspension from a fluid sample (porous) in the chamber of the PA 124 may be electroporated for a period of time, depending on the cell concentration in the cell suspension in a certain volume of the fluid sample. The flow-through electroporation configuration can set the instrument output 122 to interface with a single PA 124 through a pair of diagonally opposite male / female banana plugs, where multiple fluid samples or pores containing cells in the cell suspension in the PA 124 can be automatically and / or autonomously electroporated, such that the processor 103 and / or the SBC 102 can command the HV module 106 to run an experimental protocol with one or more bursts (each burst having some microsecond quantity), and the bursts are separated by a time pause for transferring the fluid of the next fluid sample inside the PA 124. An experimental protocol with one or more microsecond bursts separated by a time pause can be a pulse width wave. Flow-through electroporation can be a faster and more efficient way to transfect a large number of cells in a cell suspension in a fluid equal to 1L within no more than 30 minutes.

[0030] The user can connect the instrumentation device to the calibration port 126 so that the processor 103 and / or the SBC 102 can simulate a desired pulse-width wave from the instrumentation device, where the HV board 112 can command the HV power supply 116 to charge at least one capacitor bank 118 to generate a required voltage amount so that the PM board 120 generates a pulse-width wave. Although there may not be any PA 124 interfacing with the instrument output 122, the HV board 112 will operate in the instrument output 122 based on the pulse-width wave from the user's instrumentation device via the HV switch 119. The calibration port 126 can be used to calibrate the system 100 once or periodically before the operation of the electroporation device. In one embodiment, the processor 103 and / or the SBC 102 can be connected to the PM board 120 via the SB 104. In another embodiment, the processor 103 and / or the SBC 102 can be connected to the PM board 120 via the connection of the SB 104 to the HV board 112. In yet another embodiment, the processor 103 and / or the SBC 102 can be connected to the PM board 120 via the 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 can be connected to the HV board 112, the HV board 112, at least one capacitor bank 118, the HV switch 119, the PM board 120, the instrument output 122, the PA 124, and the calibration port 126 via the connection of the SB 104 to the HV module 106.

[0031] The LV module 108 can 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 3.0 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 can constitute components of the LV module 108.

[0032] The SB 104 can use one or more DC relays ( Figure 1 not shown in the figure) to gate the isolated LV DC power supply 114 from the PEM 110. In another embodiment, the isolated LV DC power supply 114 can use one or more DC relays from the LV module 108 ( Figure 1The door selects PEM 110 (not shown in the figure). In one embodiment, the processor 103 and / or the SBC 102 can manage, control, and / or collect data from 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 via the connection of the SB 104. In another embodiment, the SB 104 can be connected to all components in the LV module 108, and the processor 103 and / or the SBC 102 can manage and / or control the isolated LV DC power supply 114, 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.

[0033] In an example embodiment, the display and touch screen 146 are used. However, other embodiments can alternatively be implemented by other known user interface devices. For example, the display and touch screen 149 can alternatively be implemented using a non-touch screen display and alternative tools for user input (such as a keyboard, a mouse, programmable soft keys, hard-coded buttons, etc.).

[0034] The processor 103 and / or the SBC 102 can collect transfection data associated with cells in the cell suspension from the fluid sample inside the chamber of the PA 124 for storage in one or more storage devices in the system 100 ( Figure 1(not shown). The processor 103 and / or the SBC 102 may allow an operator or user to manipulate data on the display and touch screen 146. The SBC 102 may provide the operator or user with a system 100 having computer capabilities, where an independent desktop or laptop computer may not be required. The SBC 102 may command, manage, and / or regulate 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. The operator or user may scan the PA124 containing the fluid sample into the barcode reader 148 for the processor 103 and / or the SBC 102 to identify and associate the required pulse width wave profile based on the experimental protocol selected by the operator or user, and thus apply it to the cells in the cell suspension from the fluid sample. The scanned information that can be read by the barcode reader 148 may be stored in one or more storage devices for later identification and association with the results in the SBC 102 and / or the manipulation data generated by the operator or user on the display and touch screen 146. The processor 103 and / or the SBC 102 may receive one or more commands from the operator or user via the display and touch screen 146 to run a specific required pulse width wave profile or experimental protocol to be applied to the cells in the cell suspension from the fluid sample via the instrument output 122.

[0035] The processor 103 and / or the SBC 102 can activate the fan 150 to cool the components in the LV module 108. The processor 103 and / or the SBC 102 can use the speaker 152 to transmit the results to the operator or user based on the input from the display and touch screen 146. The processor 103 and / or the SBC 102 can detect via the SB 104 that the operator or user may have pressed the power button 154, such that the processor 103 and / or the SBC 102 can power on the system 100. The operator or user can store the saved results from the SBC 102 and / or the manipulation data generated on the display and touch screen 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 can be illuminated in one or more colors, such as blue, yellow, red, green, or orange, via the processor 103 and / or the SBC 102 using the LED board 158, where a first color can indicate that the system 100 is "on", and a second color can indicate that the system 100 is "off". Additionally, the processor 103 and / or the SBC 102 can use the LED board 158 to illuminate a hollow rectangular-shaped section on the perimeter of the instrument output 122 in one or more colors, such as blue, yellow, red, green, or orange. In another embodiment, the processor 103 and / or the SBC 102 can illuminate a hollow rectangular-shaped section on the perimeter of the instrument output 122 in one or more colors, such as blue, yellow, red, green, or orange. The color of the hollow rectangular-shaped section around the perimeter of the instrument output 122 can indicate to the operator or user the progress or status of the electroporation process. Additionally, the processor 103 and / or the SBC 102 can change the color of the hollow rectangular-shaped section on the perimeter of the instrument output 122.

[0036] The processor 103 and / or the SBC 102 can determine the type of fluid and the viscosity of the fluid in the chamber of the PA124, where a large number of cells in the cell suspension can indicate a high viscosity level in the fluid, which may require a large pressure to move the fluid inside the chamber of the PA124 during flow-through electroporation. The processor 103 and / or the SBC 102 can 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 can be used by circulating air around the chamber of the PA124 to generate a positive air pressure or a negative air pressure to move or draw a fluid sample into and out of the chamber of the PA124. For example, the inlet pump 160 can be used to generate a negative air pressure (or vice versa) by sucking air out of the chamber in the PA124 (or vice versa), which in turn can draw the fluid sample from the filled 1L bag into the chamber in the PA124. After the electroporation process can be run, the outlet pump 164 can generate a positive air pressure (or vice versa) by blowing air out of the chamber in the PA124 (or vice versa), which in turn can draw the fluid sample from the chamber in the PA124 into an empty or filling 1L bag that contains the transfected cells in the cell suspension in the fluid sample.

[0037] The processor 103 and / or the SBC 102 can use the inlet valve 162 or the outlet valve 166. Based on the processor 103 and / or the SBC 102 identifying the type and viscosity of the fluid, the processor 103 and / or the SBC 102 can precisely control the amount of fluid that circulates from the filled 1L bag into the chamber of the PA124 and out into the empty or filling 1L bag. For example, even if the inlet pump 160 and / or the outlet pump 164 may be applying a large positive and / or negative air pressure, in cooperation with the inlet pump 160 and the outlet pump 164, the processor 103 and / or the SBC 102 can actuate the inlet valve 162 and the outlet valve 166 to prevent the fluid from flowing from the filled 1L bag to the chamber of the PA124 and then to the empty or filling 1L bag, because the inlet valve 162 and the outlet valve 166 can squeeze one or more tubes connected to the PA124 to prevent fluid transfer. Even if the inlet pump 160 and / or the outlet pump 164 may be applying a low level of positive and / or negative air pressure, the processor 103 and / or the SBC 102 can accelerate or slow down the fluid flow from the filled 1L bag to the chamber of the PA124 and then to the empty or filling 1L bag, because the inlet valve 162 and the outlet valve 166 can squeeze or release one or more tubes connected to the PA124 for decelerating or accelerating fluid transfer.

[0038] An operator or user can temporarily press the emergency stop (E-stop) button 168 (referred to herein as the E-stop button 168) to immediately stop the electroporation process or the pulse width wave (which can be generated by the PM board 120 via the HV switch 119 and / or run by the HV board 112), where the D flip-flop in SB 104 ( Figure 1 not shown) can override, disable, block, and / or bypass the processor 103 and / or the SBC 102, and directly turn off or terminate the HV board 112, the HV switch 119, and the PM board 120 in the HV module 106. The D flip-flop can override, disable, block, and / or bypass the processor 103 and / or the SBC 102 to manage and / or control the HV module 106 and its components, and the D flip-flop can still utilize the processor 103 and / or the SBC 102 to manage and / or control the LV module 108 and its components. In addition, the D flip-flop in SB 104 can communicate with the processor 103 and / or the SBC 102 via the USB 3.0 port of the SBC 102 to display on the display and touch screen 146 the status that the emergency stop button 168 may have been pressed by the operator and user. The emergency stop 168 can override, disable, block, and / or bypass the processor 103 and / or the SBC 102 to immediately terminate any electroporation in the HV module 106. In addition, the emergency stop 168 can be implemented to stop the high voltage during the electroporation process for an emergency situation, where near-instantaneous deactivation can be advantageous. For example, the emergency stop 168 can be used to prevent cells in the cell suspension from the fluid sample from being damaged, prevent the system 100 from being damaged, prevent any leaked fluid from electrocuting the operator or user, or prevent an explosion of a flammable fluid sample. In addition, when overriding, disabling, blocking, and / or bypassing the processor 103 and / or the SBC 102, the emergency stop 168 and the D flip-flop in SB 104 can operate to immediately terminate the electroporation process in the HV module 106 without relying on the software on the system 100. The emergency stop button 168 can be illuminated by the processor 103 and / or the SBC 102 with a red LED light.

[0039] Figure 2 is a schematic block diagram showing an exemplary non-software-based emergency stop feature consistent with the disclosed embodiments. The system 200 can include an emergency stop button 202 (also referred to as Figure 1 the emergency stop button 168 in Figure 1 the emergency stop button 168 in Figure 2 can be the same as the emergency stop button 202 in Figure 1 the SB 104 in Figure 1 the PM board 120 inFigure 1 the HV switch 119) in, the SBC 216 (also referred to as Figure 1 the SBC 102) in, and the display and touch screen 218 (also referred to as Figure 1 the display and touch screen 146) in, all of which can form components of the system 200. The system 200 can be Figure 1 a sub-component of the system 100 in. The SB 204, D flip-flop 206, gate 208, and / or pulse shaping circuit 210 can be electronic circuits.

[0040] The emergency stop button 202 is connected to the D flip-flop 206. The D flip-flop 206 can be located inside the SB 204. The D flip-flop 206 can include a preset input pin input pin (D), clock input pin (CLK), clear input pin power pin (V CC ), output pin (Q), inverted output pin and ground pin (GND). To avoid relying on the clock characteristics of the D flip-flop 206, the input pin (D) can be connected to ground so that the clock rising edge function of the clock input pin (CLK) can be used to reset the D flip-flop 206, the ground pin (GND) can be connected to ground, and the power pin (V CC ) can be connected to the voltage level (+V) required to operate the D flip-flop 206, and the clear input pin can also be connected to a logic high level and thus be disabled.

[0041] In addition, the emergency stop button 202 can be connected to the preset input pin of the D flip-flop 206 such that when the user can instantaneously press the emergency stop button 202, a transient signal can pass through the preset input pin and be latched (the transient signal can be held, saved, and / or retained by the D flip-flop 206), where the preset input signal can have a logic value of "1". In other embodiments, the preset input signal can be a low voltage or a ground connection. The preset input pin can be asynchronous and can react immediately when the preset input signal can be pulled low (assigned a logic value of "0"), or can be pulled high (assigned a logic value of "1"), which is different from the traditional input pin (D) (not connected to ground but to an input signal), and the traditional input pin (D) can only react on the rising edge of the clock signal on the clock input pin (CLK). Even if the operator or user subsequently presses the emergency stop button 202, the preset input pin can be assigned a logic value of "1", where the system 200 can latch the signal received from the emergency stop button 202.

[0042] In some embodiments, it may be advantageous for system 200 to output the latch signal via two separate outputs, where the first output may be a hardware security mechanism and the second output (also referred to as the logic signal) may be a software security mechanism. The latch signal for the software security mechanism (the second output) may be directly output to the output pin (Q). The latch signal for the hardware security mechanism (the first output) may be directly output to the inverted output pin In the hardware security mechanism, system 200 may rely only on D flip-flop 206 to directly deactivate the power in HV module 106 ( Figure 1 as shown), where D flip-flop 206 may also override, disable, block, and / or bypass the logic signals from processor 103 and / or SBC 216 via SB 204. The advantage of the hardware security mechanism may be that it does not rely on software commands and / or logic signals (which may be vulnerable to timing or logic errors (software delays) in processor 103 and / or SBC 216), which can prevent the immediate deactivation (electroporation) of the power in HV module 106, where a combination of positive pulse profiles or waves and negative pulse profiles or waves (equivalent to, resulting in, and / or equal to the required voltage amount) may not be immediately applied to the chamber in PA124 (which contains cells in a cell suspension from a fluid sample) via the chain of PM board 212, HV switch 214, and instrument output 122 ( Figure 1 as shown). In the software security 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 the operator or user via the display and touchscreen 218 that the electroporation process may have terminated, or D flip-flop 206 may receive an input from the operator or user via processor 103 (controlled by SBC216) from the display and touchscreen 218 requesting reactivation of the electroporation process. In one embodiment, D flip-flop 206 may receive an input from the operator or user via SBC 216 from the display and touchscreen 218 requesting reactivation of the electroporation process. In another embodiment, D flip-flop 206 may receive an input from the operator or user via processor 103 from the display and touchscreen 218 requesting reactivation of the electroporation process. The software security mechanism may be vulnerable to timing or logic errors (software delays) in the logic signals of processor 103 and / or SBC 216 for the operation of the components in LV module 108 and / or HV module 106, but those software delays may not inadvertently affect D flip-flop 206 from directly deactivating, overriding, disabling, blocking, and / or bypassing the power for electroporation in HV module 106.

[0043] From a preset input pin The latch signal can be directly output as a logic value "1" to the output pin (Q), which can be transmitted to the SBC board 216, where the processor 103 can cause the display and touch screen 218 to display a message to the operator or user that the emergency stop feature can be activated. From the preset input pin The latch signal can be directly output as a logic value "0" to the inverted output pin which can be sent to the HV module 106 and / or the PM board 212 and the HV switch 214 to stop any operation and / or generation of the pulse width wave at the instrument output 122, where the D flip-flop 206 can directly deactivate, override, disable, block, and / or bypass the processor 103 and / or the SBC 216, preventing the processor 103 and / or the SBC 216 from managing and / or controlling Figure 1 the HV module 106 and its components therein. In another embodiment, the gate 208 can have a latch signal from the preset input pin with a logic value "0" as the first input, and a second input "1" from the pulse shaping circuit 210. The first input "0" coupled to the second input "1" can cause the gate 208 to output a logic value "0" to the PM board 212. The output logic value "0" from the gate 208 to the PM board 212 can disable or stop the HV switch 214 from applying the generated positive and / or negative pulse profiles or waves from the PM board 212 to Figure 1 the instrument output 122. In yet another embodiment, the output logic value "0" from the gate 208 to the PM board 212 can disable or stop the PM board 212 from generating positive and / or negative pulse profiles or waves, and the HV switch 214 can not apply any voltage to the instrument output 122.

[0044] The display of the message on the display and touch screen 218 can request the user to reset the latch signal from the emergency stop 202 back to "0" by pressing a graphical user interface (GUI) icon on the display and touch screen 218. The logic value "1" of the signal from the user's request to reset the emergency stop button 202 can be sent to the clock input pin (CLK), which is coupled to the input pin (D) that is always set to the logic value "0", and the clock input pin (CLK) can reset the preset input pin to "0". In one embodiment, the signal from the user's request to reset the emergency stop button 202 can be received by the display and touch screen 218, sent to the SBC 216 and / or the processor 103, and sent from the SBC 216 and / or the processor 103 to the clock input pin (CLK). At the preset input pin In the case of being reset to the logical value “0”, the output pin (Q) can have the logical value “0”, which can indicate to the SBC 216 and / or the processor 103 that the emergency stop button 202 may not be pressed, and normal operation can be performed as Figure 1 described therein. Additionally, in response to the preset input pin being reset to the logical value “0”, the inverted output pin in the D flip-flop 206 can have the logical value “1”. The inverted output pin with the logical value “1” can allow the processor 103 and / or the SBC 216 to command the PM board 212 to generate a pulse-width wave run by the HV switch 214, where the D flip-flop 206 no longer deactivates, overrides, disables, blocks, and / or bypasses the processor 103 and / or the SBC 216 due to the user requesting to reset the emergency stop button 202. In another embodiment, the gate 208 can receive the logical value “1” from the inverted output pin at its first input during normal operation, and the gate 208 can receive the logical value “1” from the pulse shaping circuit 210 at its second input, which can cause the gate 208 to output the logical value “1” to the PM board 212. In the case where the output is the logical value “1” from the gate 208, the PM board 212 and the HV switch 214 can resume their normal operation, where the processor 103 and / or the SBC 216 can command the PM board 212 to generate a pulse-width wave run by the HV switch 214, where the D flip-flop 206 no longer overrides, disables, blocks, and / or bypasses the processor 103 and / or the SBC 216 due to the user requesting to reset the emergency stop button 202.

[0045] Figure 3 Depicts an illustration of a graphical user interface of a display and a touch screen for resetting an emergency stop signal consistent with the disclosed embodiments. Figure 2 On the display and touch screen 218 (also referred to as Figure 1 the display and touch screen 146) of the GUI 300 can display a message 302 that reads “Reset Emergency Stop Signal?”. The operator or user can choose to touch or press (306) the GUI icon 304, which can cause the emergency stop 202 to be reset back to “0” when the user touches or presses the GUI icon 304. The logical value “1” of the signal from the user's request to reset the emergency stop button 202 can be sent to the clock input pin (CLK), which is coupled to the input pin (D) that is always set to the logical value “0” or grounded, and the clock input pin (CLK) can reset the preset input pin

[0046] Figure 4Exemplary illustrations of an integrated electroporation system and its components in static electroporation and flow electroporation configurations consistent with the disclosed embodiments. Figure 4 Views 402A and 402B of system 400 (also referred to as system 100 and system 200). In view 402A, system 400 may have a bezel subassembly 404, which has a "T" shape, with a thick and long horizontal section and a thin and short vertical section. The bezel subassembly 404 may include a display and touchscreen 406 (also referred to as Figure 1 the display and touchscreen 146 in Figure 2 and 218 in Figure 1 ), and an instrument output 408 (also referred to as Figure 1 the instrument output 122 in Figure 1 ), at the center of its thin and short vertical section. The instrument output 408 may be surrounded on its perimeter by a hollow rectangular-shaped section 410, which may be illuminated by Figure 4 the LED board 158 of Figure 1 The instrument output 408 in view 402A may be configured for static electroporation, where the female "T-slot" 412 on the instrument output 408 may interface with the male "T-slot" on PA124 in Figure 4 (not shown in

[0047] In Figure 4 View 402B of Figure 1 ), system 400 may have a bottom trim subassembly 420 that interfaces with the lower peripheral edge or profile of the bezel subassembly 404. Looking from view 402A, the bottom trim subassembly 420 may include a barcode reader 422 (also referred to as Figure 1 the barcode reader 148 of Figure 2 and 202 in Figure 1Power button 154), USB 3.0 port 428 (also referred to as Figure 1 USB 3.0 port 156) of, inlet pump 430 with knob 432 for attaching the tube of PA124 in the flow-through electroporation configuration (also referred to as Figure 1 inlet pump 160) of, inlet valve 434 (also referred to as Figure 1 inlet valve 162) of, outlet pump 436 with knob 438 for attaching the tube of PA124 in the flow-through electroporation configuration (also referred to as Figure 1 outlet pump 164) of, and outlet valve 440 (also referred to as Figure 1 outlet valve 166) of.

[0048] In Figure 4 View 402A of, system 400 may have a left panel 442, and the left panel 442 interfaces with the thick and long section of the bezel subassembly 404 and the left edge or profile of the lower trim subassembly 420. The left panel 442 may include a left hook assembly 444 in a stowed configuration. In Figure 4 View 402B of, system 400 may have a right panel 446 that interfaces with the thick and long section of the bezel subassembly 404 and the right edge or profile of the lower trim subassembly 420. The right panel 448 may include a right hook assembly 448 in a stowed configuration.

[0049] The emergency stop button 424 in view 402A may be located at the upper right corner of the lower trim subassembly 420 in view 402B, aligned with the mother "T-slot" 412. In other embodiments, the emergency stop button 424 may be located in a similar position, such as an equivalent position at the left corner of the lower trim subassembly 420. The placement of the emergency stop button 424 in this position or a similar position may have certain advantages. For example, the placement of the emergency stop button 424 may allow for easy access without damaging one or more tubes of PA124 and away from any potential leaks of the pipeline and / or PA124. This placement may also position the emergency stop button 424 closer to the operator's or user's line of sight, making it easier to identify the emergency stop button 424 in an emergency.

[0050] Figure 5 Is an exemplary illustration of a processing component that interfaces with the instrument output in a static electroporation configuration, consistent with the disclosed embodiments. Figure 5 Shows a close-up view of system 500 (also referred to as Figure 1 system 100 of, Figure 2 200 of, and Figure 4 400 of), where PA502 (also referred to as Figure 1 PA 124 of) may be configured for static electroporation. PA 502 may interface with instrument output 504 (also referred to as Figure 1The instrument output 122 and Figure 4 the 408) interface, where Figure 4 the female "T-slot" 412 in it can be connected to the male "T-slot" interface of the instrument output 504 ( Figure 5 not shown in it). The hollow rectangular section 506 (also referred to as Figure 4 the hollow rectangular section 410) on the periphery of the instrument output 504 can be Figure 1 illuminated by the LED board 158 to indicate to the operator or user that the system 500 may be in a static electroporation configuration.

[0051] Figure 6 is an exemplary illustration of a processing component that interfaces with the instrument output in a flow-through electroporation configuration, consistent with the disclosed embodiments. Figure 6 The system 600 with views 602A, 602B, and 602C is shown (also referred to as Figure 1 the system 100, Figure 2 the 200, Figure 4 the 400, and Figure 5 the 500). Figure 6 View 602A of it includes a bezel sub-component 604 (also referred to as Figure 4 the bezel sub-component 404), which has a display and a touch screen 606 (also referred to as Figure 1 the display and touch screen 146, Figure 2 the display and touch screen 218, and Figure 4 the display and touch screen 406), and a hollow rectangular section 608 (also referred to as Figure 6 the hollow rectangular section 410 and Figure 4 the hollow rectangular section 410 and Figure 5 the hollow rectangular section 506) on the periphery of the instrument output 504 (

[0052] Figure 6 View 602A in it shows a lower trim sub-component 610 (also referred to as Figure 4 the lower trim sub-component 420), which includes an inlet pump 612 (also referred to as Figure 1 the inlet pump 160 and Figure 4 the inlet pump 430), an inlet valve 614 (also referred to as Figure 1 the inlet valve 162 and Figure 4 the inlet valve 434), an outlet pump 616 (also referred to as Figure 1 the outlet pump 164 and Figure 4 the outlet pump 436) and an outlet valve 618 (also referred to as Figure 1 the outlet valve 166 and Figure 4 the outlet valve 440).

[0053] In one embodiment, the PA configured for flow-through electroporation may include a filled 1L bag 620, an inlet tube 622, a chamber 624, an inlet / discharge tube 626, an inlet / discharge tube 628, an outlet tube 630, and an empty or filling 1L bag 632. The filled 1L bag 620 may be connected to the inlet tube 622, and the inlet tube 622 may be connected to the inlet valve 614. The inlet tube 622 may enter the chamber 624, and the inlet / discharge tube 626 may leave the chamber 624 and may be connected to the inlet pump 612. The inlet / discharge tube 626 may leave the inlet pump 612, and the end orifice of the inlet / discharge tube 626 may be exposed to air.

[0054] In addition, the inlet / discharge tube 628 may leave the chamber 624 and may be connected to the outlet pump 616. The inlet / discharge 628 may leave the outlet pump 616, and the end orifice of the inlet / discharge 628 may be exposed to air. Further, the outlet tube 630 may leave the chamber 624 and be connected to the outlet valve 618. The outlet tube 630 may leave the outlet valve 618 to connect to the empty or filling 1L bag 632.

[0055] During operation, the inlet pump 612 may pressurize the inlet / discharge tube 626, the chamber 624, and the inlet tube 622 to blow air out of the end orifice of the inlet / discharge tube 626, which may cause fluid from the filled 1L bag 620 to flow through the inlet tube 622 (regulated by valve 614) and into the chamber 624. Similarly, the outlet pump 612 may pressurize the inlet / discharge tube 628, the chamber 624, and the outlet tube 630 to blow air out of the end orifice of the inlet / discharge tube 628, which may cause the fluid inside the chamber 624 to flow through the outlet tube 630 (regulated by valve 618) and into the empty or filling 1L bag 632. The fluid in the chamber 624 may be electroporated by the instrument output 504. A pair of diagonally opposite male banana plugs from the chamber 624 ( Figure 6 not shown in) may interface with a pair of diagonally opposite female banana plugs 418 on the instrument output 504 ( Figure 6 not shown in). The SB 204 may manage and coordinate the flow of fluid in the chamber 624 by controlling the inlet pump 612, the inlet valve 614, the outlet pump 616, and the outlet valve 618. For example, the SB 204 may cause the inlet pump 612 and the outlet pump 616 to suck air through the end orifice of the inlet / discharge tube 626 and the end orifice of the inlet / discharge tube 628, respectively, to prevent fluid from entering or leaving the chamber 624 during electroporation, where the inlet valve 614 may prevent fluid from flowing in the inlet tube 622, and the outlet valve 618 may prevent fluid from flowing in the outlet tube 630. The SB 204 may determine the correct amount of fluid for electroporation in the chamber 624 via the current sensor 134.

[0056] Figure 6 View 602B therein shows the left panel 634 (also referred to as Figure 4 the left panel 442 of Figure 4 ), where the left hook assembly 636 (also referred to as Figure 6 the left hook assembly 444 of Figure 4 ) is in the deployed configuration, where the left hook assembly 636 can support the filled 1L bag 620 in View 602A. Figure 4 View 602C therein shows the right panel 638 (also referred to as

[0057] Figure 7 the left panel 446 of Figure 7 ), where the right hook assembly 640 (also referred to as Figure 1 the left hook assembly 448 of Figure 1 ) is in the deployed configuration, where the right hook assembly 640 can support the empty or being-filled 1L bag 632 in View 602A. Figure 4 the bezel subassembly 404 of Figure 6 and Figure 4 the lower fascia subassembly 420 of Figure 6 and Figure 1 the display and touch screen 146 of Figure 2 214 of Figure 4 406 of Figure 6 and Figure 1 the SBC 102 of Figure 2 and Figure 1 the barcode reader 148 of Figure 4 and Figure 1 the instrument output 122 of Figure 4 408 of Figure 5 and Figure 1 the LED board 158 of Figure 1 the outlet pump 164 of Figure 4 436 of Figure 6 and Figure 1 the outlet valve 166 of Figure 4 440 of Figure 6618), inlet pump 722 (also referred to as Figure 1 inlet pump 160 of Figure 4 430 of Figure 6 612), inlet valve 724 (also referred to as Figure 1 inlet valve 162 of Figure 4 434 of Figure 6 614), power button 726 (also referred to as Figure 1 power button 154 of Figure 4 426), USB 3.0 port 728 (also referred to as Figure 1 USB 3.0 port 156 of Figure 4 428), and emergency stop 730 (also referred to as Figure 1 emergency stop button 168 of Figure 2 202 of Figure 4 424).

[0058] Facing the front board assembly as shown in Figure 7 , the bezel sub - assembly 702 and the lower trim board sub - assembly 704 can be mounted on the front board assembly 706. The display and touchscreen 708 can be mounted to the bezel sub - assembly 702. The SBC 710 can be mounted on the upper right side of the front board assembly 706. The barcode reader 712 can be mounted on the upper right of the lower trim board sub - assembly 704 and can be mounted below the SBC 710 on the front board assembly 706. The instrument output 730 can be mounted on the bezel sub - assembly 702. The LED board 716 can be mounted on the instrument output 730 facing the front board assembly 706. The outlet pump 718 can be mounted at the lower left of the lower trim board sub - assembly 704 and the lower left of the front board assembly 706. The outlet valve 720 can be mounted at the center of the lower trim board sub - assembly 704 on the right side of the outlet pump 718. The inlet pump 722 can be mounted at the lower right of the lower trim board sub - assembly 704 and the lower right of the front board assembly 706. The inlet valve 724 can be mounted at the center of the lower trim board sub - assembly 704, to the left of the inlet pump 722 and to the right of the outlet valve 720. The power button 726 can be mounted at the center of the bottom of the lower trim board sub - assembly below the outlet valve 720. The USB 3.0 port 728 can be mounted at the center of the lower trim board sub - assembly 704 below the inlet valve 724 and mounted to the right of the power button 726. The emergency stop button 730 can be mounted at the upper left of the lower trim board sub - assembly 704.

[0059] Figure 8 An exemplary block diagram showing an exemplary method for electroporating cells in a cell suspension in a fluid in accordance with some embodiments of the present disclosure. As Figure 8As shown, at block 802, method 800 may be executed by the system board, which may receive a signal from the emergency stop button 168 to stop the voltage applied to the cells in the cell suspension in the fluid by the instrument output 122. The signal may be a transient signal generated at the instant the button of the other signal device is activated and is used as an interruption to the electroporation process on the cells in the cell suspension of the fluid.

[0060] At block 804, method 800 may be executed by the system board, which may latch the signal from the emergency stop button 168. Once the transient signal is input, the signal may be latched without waiting for a clock signal. The signal may remain latched even if the input transient signal is no longer received.

[0061] At block 806, method 800 may be executed by the system board, which may block the logic signal that can control the voltage delivery to the instrument output. The at least one processor may send a logic signal to control the voltage delivery to the instrument output.

[0062] At block 808, method 800 may be executed by the system board, which may terminate the voltage applied to the cells in the cell suspension in the fluid by the instrument output 122. The termination may occur almost instantaneously without waiting for further input from the processor 103.

[0063] At block 810, method 800 may be executed by the system board, which may disable the processor 103 from providing instructions to apply a voltage to the cells in the cell suspension in the fluid through the instrument output 122 to the high voltage module 106.

[0064] At block 812, method 800 may be executed by the processor 103, which may send a message to the user via the display and touch screen device 146 on the low voltage module 108 to reset the signal from the emergency stop 168. The signal may be sent after block 808 is run but is independent of block 810. Block 808 and block 810 may occur in parallel.

[0065] Although the present disclosure has been shown and described with reference to particular embodiments thereof, it should be understood that the present disclosure may be practiced in other environments without modification. The foregoing description has been presented for purposes of illustration. It is not exhaustive and is not limited to the precise forms or embodiments disclosed. Modifications and adaptations will be apparent to those skilled in the art in light of the specification and practice of the disclosed embodiments. Additionally, although aspects of the disclosed embodiments are described as being stored in memory, those skilled in the art will appreciate that these aspects may also be stored on other types of computer-readable media, such as secondary storage devices, e.g., hard disks or CD ROMs, or other forms of RAM or ROM, USB media, DVDs, Blu-ray or other optical drive media.

[0066] A computer program based on the written description and the disclosed method is within the capabilities of an experienced developer. Various programs or program modules may be created using any technology known to those skilled in the art, or various programs or program modules may be designed in conjunction with existing software. For example, a design segment or program module may be designed in or by means of the.Net framework,.Net Compact Framework (and related languages, such as Visual Basic, C, etc.), Java, C++, Objective-C, HTML, HTML / AJAX combination, XML, or HTML including Java applets.

[0067] Moreover, although illustrative embodiments have been described herein, those skilled in the art will understand the scope of any and all embodiments having equivalent elements, modifications, omissions, combinations (e.g., combinations of aspects across various embodiments), adaptations, and / or alterations based on the present disclosure. The limitations in the claims will be broadly construed based on the language employed in the claims and are not limited to the examples described in the specification or during the prosecution of the present application. These examples should be construed as non-exclusive. Additionally, the steps of the disclosed method may be modified in any manner, including by reordering steps and / or inserting or deleting steps. Accordingly, the specification and embodiments are to be considered illustrative only, and the true scope and spirit are indicated by the full scope of the appended claims and their equivalents.

Claims

1. An apparatus for electroporating cells in a cell suspension in a fluid, the apparatus comprises: a memory that stores instructions; a high-voltage module that includes an instrument output configured to apply a voltage to cells in a cell suspension in a fluid; a low-voltage module that includes 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 that 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 instrument output; and an emergency stop button connected to the system board; wherein the system board further includes an electronic circuit configured to run the 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 instrument output; latching the signal from the emergency stop button; blocking a logic signal that controls voltage delivery to the instrument output; terminating the voltage applied to the cells in the cell suspension in the fluid by the instrument output; and disabling the at least one processor from providing commands to the high-voltage module.

2. The apparatus according to claim 1, wherein the high-voltage module includes a high-voltage board and a pulse modulator board configured to run a positive pulse wave in the instrument output.

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

4. The apparatus according to claim 1, wherein the high-voltage module includes a high-voltage board and a pulse modulator board configured to run a negative pulse wave in the instrument output.

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

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

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

8. The apparatus according to claim 1, wherein the high-voltage module includes a calibration port configured to simulate positive and negative pulse waves in the instrument output.

9. The apparatus according to claim 1, wherein The electronic circuit includes a D flip-flop, which is connected 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 instrument output via an inverted output pin.

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

11. A method for performing safety steps to terminate the electroporation of cells in a cell suspension in a fluid, the method comprises: receiving a signal from an emergency stop button to stop the voltage applied to the cells in the cell suspension in the fluid by the instrument output; latching the signal from the emergency stop button; blocking a logic signal that controls the voltage delivery to the instrument output; terminating the voltage applied to the cells in the cell suspension in the fluid by the instrument output; and disabling the at least one processor from providing commands to the high-voltage module.

12. The method according to claim 11, wherein, the high-voltage module includes a high-voltage board and a pulse modulator board to run a positive pulse wave in the instrument output.

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

14. The method according to claim 11, wherein, the high-voltage module includes a high-voltage board and a pulse modulator board to run a negative pulse wave in the instrument output.

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

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

17. The method according to claim 11, wherein, the high-voltage module includes a calibration port configured to simulate positive and negative pulse waves in the instrument output.

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

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

20. A device for electroporating cells in a cell suspension in a fluid, the device comprises: a memory for storing instructions; A high-voltage module, the 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, the 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, 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 by the instrument output to the cells in the cell suspension in the fluid; An emergency stop button connected 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 connected to the low-voltage module, a clock input pin, and an inverted output pin connected to the high-voltage module; and wherein the D flip-flop is configured to execute instructions to perform steps including the following: Receiving a signal from the emergency stop button to stop the voltage applied by the instrument 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 voltage delivery to the instrument output; Terminating the voltage applied by the instrument output to the cells in the cell suspension in the fluid; and Disabling the at least one processor from providing commands to the high-voltage module.

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