Electroporation pipettors, systems, and methods of use thereof

Abstract

An electroporation system includes one or more of a pipette, a pipette tip, a pipette docking assembly, and a pulse generator. The pipette docking assembly includes a pipette station, a pipette station guard, and a reservoir (e.g., a buffer tube). A method for transfecting a cell with a payload includes providing an electroporation system, providing the cell, providing the payload, introducing the cell and the payload into a pipette tip, and electroporating the cell within the pipette tip by operating the electroporation system.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 408,032, filed on September 19, 2022, the disclosure of which is considered part of the disclosure of this application and is incorporated by reference in its entirety into the disclosure of this application. Background Art Technical Field

[0003] The present invention relates generally to cell transfection, and more particularly to pipettes, pipette tips, assemblies, electroporation systems, and methods for transfecting cells. Background of the Invention

[0005] Some electroporation systems include a pipette for containing target cells and a payload (e.g., a nucleic acid and / or protein to be introduced into the target cells) and an electric pulse generator for providing electric pulses to the target cells. The pipette can be connected to or inserted into a docking station associated with the electric pulse generator so that the electric pulses generated by the electric pulse generator can reach the target cells.

[0006] For example, a pipette electrode that is conductively connected to one end of the pipette chamber of the pipette (e.g., accommodating target cells and payload) can be docked with a first electrode on a docking station. The pipette tip (e.g., including the open end of the pipette chamber) can be inserted into a buffer solution that is conductively connected to a second electrode on the docking station (e.g., in a reservoir), thereby exposing the open end of the pipette chamber to the buffer solution. With the pipette so connected to the docking station, an electrical pulse generator can provide electrical pulses to the first and second electrodes of the docking station, thereby allowing the electrical pulses to travel through the pipette chamber to reach the target cell and electroporate the target cell.

[0007] There are numerous disadvantages with existing electroporation systems and system components (eg, pipettes, pipette tips, pipette docking assemblies, and pulse generators), and there is a continuing need and desire for improved electroporation systems including improved components. Summary of the invention

[0008] Various methods of the present disclosure extend at least to electroporation systems, components thereof, and / or methods associated therewith.

[0009] In one aspect, the invention provides an electroporation system. In embodiments, the system includes one or more of a pipette, a pipette tip, a pipette docking assembly, and a pulse generator. In some embodiments, the pipette docking assembly includes a pipette station, a pipette station guard, and a reservoir.

[0010] On the other hand, the present disclosure provides a pipette. In an embodiment, the pipette includes: a proximal section having a handle; a distal section configured to be reversibly attached to a pipette tip; a first actuator disposed in the proximal section, which, when actuated, is operable to control: i) the pipetting function of the pipette; and ii) the gripping and release of a plunger disposed in an inner cavity of the pipette tip; and a second actuator disposed in the proximal section, which, when actuated, is operable to separate the pipette tip from the distal section of the pipette. In various embodiments, the pipette includes a pipette electrode disposed in the distal section, which is electrically coupled to the plunger when the plunger is operably coupled to the first actuator.

[0011] On the other hand, the present disclosure provides a pipette tip that is configured to be reversibly attached to a pipette. In an embodiment, the pipette tip includes: a tip sleeve defining an inner cavity extending from the proximal end of the pipette tip to the distal end of the pipette tip; a plunger at least partially disposed in the inner cavity, the plunger being composed of a conductive material and being configured to translate along the inner cavity to facilitate aspiration of the fluid into the inner cavity and / or dispensing the fluid from the inner cavity; and an attachment interface disposed at the proximal end of the pipette tip, the attachment interface including one or more tabs configured to engage with the pipette. In some embodiments, the attachment interface includes one or more tabs configured to engage with a holding platform of a distal segment of the pipette. In some embodiments, one or more tabs are configured to interact with a biasing member of the pipette during attachment of the pipette tip to the pipette via a holding platform. In various embodiments, the pipette tip has a sample volume capacity between 10 μL and 100 μL.

[0012] In various aspects, the present disclosure provides a pipette assembly including a pipette reversibly attached to a pipette tip. In some embodiments, the pipette of the assembly includes a proximal segment with a handle, a distal segment with a tip interface, and a first actuator disposed in the proximal segment. The pipette tip reversibly attached to the pipette of the assembly includes: a tip sleeve defining an inner cavity extending from the proximal end of the pipette tip to the distal end of the pipette tip; a plunger at least partially disposed in the inner cavity; and an attachment interface disposed at the proximal end of the pipette tip. In various embodiments, the plunger is reversibly operably coupled to the first actuator, and when operably coupled, performs a pipetting function when the first actuator is actuated by translating along the inner cavity. In addition, the tip sleeve is reversibly attached to the distal segment via one or more tabs of the attachment interface that engage with the retention platform of the distal segment. In an embodiment, the pipette of the assembly further comprises a pipette electrode disposed in the distal segment and electrically coupled to the plunger when the plunger is operably coupled to the first actuator.

[0013] In some embodiments, the pipette of the assembly includes a proximal section with a handle, a distal section with a tip interface, a first actuator disposed in the proximal section, and a clamp mechanism with a clamp jaw disposed in the distal section. The pipette tip reversibly attached to the pipette of the assembly includes: a tip sleeve defining an inner cavity extending from the proximal end of the pipette tip to the distal end of the pipette tip; and a plunger at least partially disposed in the inner cavity. In various embodiments, the plunger is reversibly operably coupled to the first actuator via the clamp jaw, and when operably coupled, performs a pipetting function when the first actuator is actuated by translating along the inner cavity. In an embodiment, the pipette of the assembly also includes a pipette electrode, which is disposed in the distal section and electrically coupled to the plunger when the plunger is operably coupled to the clamp jaw (e.g., grasped by the clamp jaw).

[0014] In another aspect, the present disclosure provides a pulse generator including one or more connection ports. Each specific connection port in the one or more connection ports includes a corresponding port door. Each corresponding port door includes a corresponding biasing element, which is used to bias the corresponding port door into a closed configuration to prevent access to the specific connection port. In some cases, each corresponding port door includes a corresponding tool interface configured to receive a port door tool. The port door tool is configured to interact with the corresponding tool interface to react to the corresponding biasing element of the corresponding port door, so that the corresponding port door is in an open configuration to provide access to the specific connection port. The insertion of the port connection component into the specific connection port can maintain the reaction to the corresponding biasing element to keep the corresponding port door in an open configuration, and the disconnection of the port connection component from the specific connection port can remove the reaction to the corresponding biasing element to allow the corresponding port door to return to the closed configuration.

[0015] In yet another aspect, the present disclosure provides a method for transfecting cells with a payload. The method comprises: providing an electroporation system of the present disclosure; providing cells; providing a payload; introducing the cells and the payload into a pipette tip of a pipette attached to the system; and electroporating the cells by operating the electroporation system. In some embodiments, the cells are mammalian cells. In some embodiments, the payload comprises a nucleic acid, a protein, or a combination thereof.

[0016] Additional features and advantages will be set forth in the following description, and in part will be apparent from the description, or may be learned by practicing the teachings herein. The features and advantages of the systems described herein may be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. Features of the invention will become more fully apparent from the following description and the appended claims, or may be learned by practicing the invention as described below. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more fully understand the nature and advantages of the present invention, reference should be made to the following detailed description in conjunction with the accompanying drawings. It should be understood that these drawings only depict typical embodiments of the present invention and therefore should not be considered as limiting the scope thereof. The present disclosure will be described and explained in more detail through the use of the accompanying drawings.

[0018] Figure 1 is a schematic diagram showing example components of an electroporation system of the present disclosure and its components in one embodiment of the present disclosure.

[0019] FIG. 2A to FIG. 2B is a schematic diagram illustrating aspects of an example pipette tip (eg, a consumable pipette tip) for use with an electroporation system in embodiments of the present disclosure. Figure 2A is a schematic diagram illustrating aspects of an example pipette tip in one embodiment of the present disclosure. Figure 2B is a schematic diagram illustrating aspects of an example pipette tip in one embodiment of the present disclosure.

[0020] FIG. 3A to FIG. 3B It is shown FIG. 2A to FIG. 2B Schematic diagram of an exploded view of a pipette tip depicted in FIG. Figure 3A yes Figure 2A Exploded view of the pipette tip depicted in . Figure 3B yes Figure 2B Exploded view of the pipette tip depicted in .

[0021] FIG. 4A to FIG. 4B is a schematic diagram illustrating aspects of a plunger of a pipette tip in an embodiment of the present disclosure. Figure 4A is an exploded view of an example plunger. Figure 4B After assembly Figure 4A Schematic diagram of the plunger depicted in FIG.

[0022] FIG. 5A to FIG. 5B is a schematic diagram illustrating aspects of a pipette tip in an embodiment of the present disclosure. Figure 5A is a cross-sectional view of a pipette tip in one embodiment of the present disclosure. Figure 5B yes Figure 5A An expanded cross-sectional view of the distal portion of the pipette tip depicted in FIG.

[0023] Figure 6 is a schematic diagram illustrating aspects of a plunger of a pipette tip in an embodiment of the present disclosure.

[0024] FIG. 7A to FIG. 7B is a schematic diagram illustrating a pipette tip in one embodiment of the present disclosure. Fig. 7A is a schematic diagram of a pipette tip of the present disclosure. Figure 7B yes Fig. 7A Exploded view of the distal portion of the pipette tip depicted in FIG.

[0025] FIG. 8A to FIG. 8B is a schematic diagram showing components of a pipette and a pipette tip in an embodiment of the present disclosure. Fig. 8A is a perspective view of a pipette in one embodiment of the present disclosure. Figure 8B In one embodiment of the present disclosure, Fig. 8A A perspective view of a pipette tip used with the pipette depicted in FIG.

[0026] 9A to 9E is a schematic diagram illustrating the interaction between a pipette and a pipette tip in an embodiment of the present disclosure. Fig.9A is a schematic diagram showing the interaction of the attachment interface of a pipette tip with the engagement interface of a pipette. Fig. 9B is a schematic diagram showing the interaction of the attachment interface of a pipette tip with the engagement interface of a pipette. Fig. 9C is a cross-sectional view of the distal end of the pipette beginning to engage the proximal end of the pipette tip. Fig.9D is a cross-sectional view of the distal end of the pipette engaging the proximal end of the pipette tip. Fig.9E is a cross-sectional view of the tip interface at the distal end of a pipette attached to the attachment interface at the proximal end of a pipette tip.

[0027] FIG. 10A to FIG. 10B is a cross-sectional view illustrating the pipetting functionality of a pipette and attached pipette tips in an embodiment of the present disclosure. Fig. 10A is a cross-sectional view showing the transition of components of a pipette and a pipette tip for aspirating fluid into a lumen of a pipette tip. Fig. 10B is a cross-sectional view showing the transition of components of a pipette and a pipette tip for dispensing fluid from an interior cavity of a pipette tip.

[0028] FIG. 11A to FIG. 11B is a schematic diagram illustrating functional and structural aspects of a pipette and a pipette tip in an embodiment of the present disclosure. Fig.11A is a schematic diagram showing the operation of the pipette. Fig. 11B is a schematic diagram showing the operation of the pipette.

[0029] Fig.12 is a perspective view showing aspects of an example pipette station guard of an electroporation system in an embodiment of the present disclosure.

[0030] FIG. 13A to FIG. 13B is shown in the embodiment of the present disclosure Fig.12 A perspective view of the assembly of the pipette station guard and the pipette station depicted in FIG. Fig.13AThe insertion and rotation of the pipette station guard during assembly with the pipette station is shown. Fig. 13B The pipette station guard is shown assembled with the pipette station, with the base of the pipette station guard pushed toward the pipette station to complete the assembly.

[0031] Fig.14 An example reservoir, also referred to as a buffer tube, of an electroporation system in an embodiment of the present disclosure is shown.

[0032] FIG. 15A to FIG. 15B is shown in the embodiment of the present disclosure Fig.14 The storage device depicted in Fig. 13B A perspective view of the assembly of the pipette station guard and pipette station is depicted in FIG. Fig.15A Alignment of the reservoir with the pipette station guard is shown for assembly with the assembled pipette station and pipette station guard. Fig. 15B The reservoir is shown assembled with the pipette station and the pipette station guard to form a fully assembled pipetting docking assembly.

[0033] Fig.16 is a cross-sectional side view of a fully assembled pipette docking assembly in an embodiment of the present disclosure.

[0034] Fig.17 is a cross-sectional side view of a fully assembled pipette docking assembly in an embodiment of the present disclosure.

[0035] FIG. 18A to FIG. 18B The pipette is assembled into Fig. 15B Depicted is a perspective view of the pipette docking assembly in the reservoir to perform the electroporation procedure. Fig.18A Alignment of the pipette with the pipette station guard is shown for assembly with the pipette docking assembly. Fig.18B The pipette is shown assembled with the pipette docking assembly.

[0036] Fig.19 is a side view illustrating aspects of the assembly of a pipette and a pipette docking assembly in an embodiment of the present disclosure.

[0037] Fig. 20 is a rear elevation view of a pulse generator showing aspects of an example port gate in an embodiment of the present disclosure.

[0038] Fig.21 is a perspective view showing aspects of components of a pulse generator in an embodiment of the present disclosure.

[0039] Fig. 22 is an expanded cutaway perspective view showing aspects of components of a pulse generator in an embodiment of the present disclosure.

[0040] Fig.23is an expanded cutaway perspective view showing aspects of components of a pulse generator in an embodiment of the present disclosure.

[0041] FIG. 24A to FIG. 24B is an expanded cutaway perspective view showing aspects of components of a pulse generator in an embodiment of the present disclosure. Fig.24A A connection component for receiving a portable drive (eg, USB) device is shown. Fig. 24B A USB device is shown plugged into the connection component.

[0042] FIG. 25A to FIG. 25B is an expanded perspective view showing aspects of components of a pulse generator in an embodiment of the present disclosure. Fig.25A A USB device is shown plugged into the connection component. Fig.25B A door covering the connection components is shown, which automatically closes when a connected device (eg, a USB device) is removed.

[0043] FIG. 26A to FIG. 26B is a perspective view showing aspects of an example cable adapter. Fig.26A Features of a cable adapter are shown. Fig.26B is an exploded view of the components of the cable adapter.

[0044] FIG. 27A to FIG. 27C is a cross-sectional view showing aspects of an example cable adapter and pulse generator. Fig.27A A cross-sectional view of the cable adapter and pulse generator is shown. Fig.27B A cross-sectional view of the attachment of a pulse generator and a cable adapter is shown with corresponding attachment features aligned. Fig.27C The mounting holes and cable adapter mounting features of the pulse generator are shown as Fig.27B Aligned as shown, a cross-sectional view of attaching the cable adapter to the pulse generator by sliding the cable adapter downward relative to the pulse generator.

[0045] FIG. 28A to FIG. 28C Aspects of an example cable tie assembly for an electroporation system are shown. Fig.28A A cable tie assembly having connection portions for electrically coupling to a pulse generator and a pipette station is shown. Fig.28B Features of a connection portion of a cable tie assembly configured to be electrically coupled to a pulse generator are shown. Fig.28C Features of a connection portion of a cable tie assembly configured to be electrically coupled to a pipette station are shown.

[0046] FIG. 29A to FIG. 29B is a schematic diagram showing aspects of a clamp for a cable assembly. Fig.29A Features of a clamp for engaging cables are shown. Fig.29B Shows Fig.29AStructural features of the assembled fixture.

[0047] Fig.30 is a schematic cross-sectional view of a pipette docking assembly having a pipette and an attached pipette tip including a sample docked within the docking assembly for performing an electroporation procedure.

[0048] Fig.31 is a graphical representation showing aspects of example electrical pulses generated by a pulse generator of an electroporation system in embodiments of the present disclosure.

[0049] Fig.32 is a graphical representation showing aspects of example electrical pulses generated by a pulse generator of an electroporation system in embodiments of the present disclosure.

[0050] Fig.33 is a schematic diagram illustrating an example circuit architecture that may be implemented in a pulse generator of an electroporation system in accordance with embodiments of the present disclosure.

[0051] Fig.34 is a schematic diagram illustrating an example circuit architecture that may be implemented in a pulse generator of an electroporation system in accordance with embodiments of the present disclosure.

[0052] Fig.35 is a graphical diagram illustrating an example electrical waveform produced by a pulse generator in an embodiment of the present disclosure.

[0053] Fig.36 is a graphical diagram illustrating an example electrical waveform produced by a pulse generator in an embodiment of the present disclosure.

[0054] Fig.37 is a graphical diagram illustrating an example electrical waveform produced by a pulse generator in an embodiment of the present disclosure.

[0055] Fig.38 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0056] Fig.39 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0057] Fig.40 is a schematic diagram illustrating aspects of the circuitry of a pulse generator in an embodiment of the present disclosure.

[0058] Fig.41 is a schematic diagram illustrating aspects of the circuitry of a pulse generator in an embodiment of the present disclosure.

[0059] Fig.42 is a schematic diagram illustrating aspects of the circuitry of a pulse generator in an embodiment of the present disclosure.

[0060] Fig.43 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0061] Fig.44 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0062] Fig.45 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0063] Fig.46 is a graphical diagram illustrating aspects of arcing and arc detection associated with an electroporation system.

[0064] Fig.47 is a front right perspective view of one embodiment of an electroporation system of the present disclosure.

[0065] Fig.48 is a front right perspective view of one embodiment of a pulse generator of the present disclosure.

[0066] Fig.49 yes Fig.48 Front elevation view of the pulse generator depicted in FIG.

[0067] Fig.50 yes Fig.48 Rear elevation view of the pulse generator depicted in FIG.

[0068] Fig.51 yes Fig.48 Right side elevation view of the pulse generator depicted in FIG.

[0069] Fig.52 yes Fig.48 Left side elevation view of the pulse generator depicted in FIG.

[0070] Fig.53 yes Fig.48 A top plan view of the pulse generator depicted in FIG.

[0071] Fig.54 yes Fig.48 A bottom plan view of the pulse generator depicted in FIG.

[0072] Fig.55 is a front right perspective view of one embodiment of a docking station assembled with a pipette station guard.

[0073] Fig.56 yes Fig.55 A front elevation view of the docking station and pipette station guard assembly depicted in FIG.

[0074] Fig.57 yes Fig.55 Rear elevation view of the docking station and pipette station guard assembly depicted in FIG.

[0075] Fig.58 yes Fig.55 A right side elevation view of the docking station and pipette station guard assembly depicted in FIG.

[0076] Fig.59 yes Fig.55 Left side elevation view of the docking station and pipette station guard assembly depicted in FIG.

[0077] Fig.60 yes Fig.55 A top plan view of the docking station and pipette station guard assembly depicted in FIG.

[0078] Fig.61 yes Fig.55 A bottom plan view of the docking station and pipette station guard assembly depicted in FIG.

[0079] Fig.62 is a front right perspective view of one embodiment of a pipette station assembled with a pipette station guard and reservoir.

[0080] Fig.63 yes Fig.62 Front elevation view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0081] Fig.64 yes Fig.62 Rear elevation view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0082] Fig.65 yes Fig.62 Right side elevation view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0083] Fig.66 yes Fig.62 Left side elevation view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0084] Fig.67 yes Fig.62 A top plan view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0085] Fig.68 yes Fig.62 A bottom plan view of the pipette station, pipette station guard, and reservoir assembly depicted in FIG.

[0086] Fig.69 is a front right perspective view of one embodiment of a reservoir.

[0087] Fig.70 yes Fig.69 A front elevation view of the reservoir depicted in FIG.

[0088] Fig.71 yes Fig.69 Rear elevation view of the reservoir depicted in FIG.

[0089] Fig.72 yes Fig.69 Right side elevation view of the reservoir depicted in FIG.

[0090] Fig.73 yes Fig.69 Left side elevation view of the reservoir depicted in FIG.

[0091] Fig.74 yes Fig.69 A top plan view of the reservoir depicted in FIG.

[0092] Fig.75 yes Fig.69 A bottom plan view of the reservoir depicted in FIG.

[0093] Fig.76 is a front right perspective view of one embodiment of a pipette.

[0094] Fig.77 yes Fig.76 Front elevation view of the pipette depicted in FIG.

[0095] Fig.78 yes Fig.76 Rear elevation view of the pipette depicted in FIG.

[0096] Fig.79 yes Fig.76 Right side elevation view of the pipette depicted in FIG.

[0097] Fig.80 yes Fig.76 Left side elevation view of the pipette depicted in FIG.

[0098] Fig.81 yes Fig.76 A top plan view of the pipette depicted in FIG.

[0099] Fig.82 yes Fig.76 A bottom plan view of the pipette depicted in FIG.

[0100] Fig.83 is a front right perspective view of one embodiment of a pipette tip having a sample volume capacity of 100 μL.

[0101] Fig.84 yes Fig.83 Front elevation view of the pipette tip depicted in FIG.

[0102] Fig.85 yes Fig.83 Rear elevation view of the pipette tip depicted in FIG.

[0103] Fig.86 yes Fig.83 Right side elevation view of the pipette tip depicted in FIG.

[0104] Fig.87 yes Fig.83 Left side elevation view of the pipette tip depicted in FIG.

[0105] Fig.88 yes Fig.83 Top plan view of the pipette tip depicted in FIG.

[0106] Fig.89 yes Fig.83 A bottom plan view of the pipette tip depicted in FIG.

[0107] Fig.90 is a front right perspective view of one embodiment of a pipette tip having a sample volume capacity of 10 μL.

[0108] Fig.91 yes Fig.90 Front elevation view of the pipette tip depicted in FIG.

[0109] Fig.92 yes Fig.90 Rear elevation view of the pipette tip depicted in FIG.

[0110] Fig.93 yes Fig.90 Right side elevation view of the pipette tip depicted in FIG.

[0111] Fig.94 yes Fig.90 Left side elevation view of the pipette tip depicted in FIG.

[0112] Fig.95 yes Fig.90 Top plan view of the pipette tip depicted in FIG.

[0113] Fig.96 yes Fig.90 A bottom plan view of the pipette tip depicted in FIG. DETAILED DESCRIPTION

[0114] Before describing in detail various embodiments of the present disclosure, it should be understood that the present disclosure is not limited to the parameters of the systems, methods, devices, components, products, processes, consumables and / or kits of the particular examples, which parameters may vary, of course. Therefore, although certain embodiments of the present disclosure will be described in detail with reference to specific configurations, parameters, components, elements, etc., these descriptions are exemplary and should not be construed as limiting the scope of the claimed invention. In addition, the terms used herein are for the purpose of describing the embodiments and are not necessarily used to limit the scope of the claimed invention.

[0115] In addition, it should be understood that for any given component or embodiment described herein, unless otherwise understood or described implicitly or explicitly, any possible candidates or alternatives listed for that component may generally be used alone or in combination with each other. In addition, it should be understood that any list of such candidates or alternatives is merely exemplary and non-restrictive unless otherwise understood or described implicitly or explicitly.

[0116] In addition, unless otherwise indicated, numbers expressing quantities, components, distances, or other measurements used in the specification and claims are to be understood as modified by the term "about," as that term is defined herein. As used herein, the terms "about" and "approximately" when referring to a measurable value such as an amount, dose, time, temperature, activity, level, quantity, frequency, percentage, dimension, size, amount, weight, position, length, etc., are intended to cover variations of ±15%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount, dose, time, temperature, activity, level, quantity, frequency, percentage, dimension, size, amount, weight, position, length, etc.

[0117] The term "comprising," which is synonymous with "including," "containing," "having," or "characterized by," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps.

[0118] It should be noted that throughout this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "an entrance" includes one, two or more entrances.

[0119] As used in the specification and the appended claims, the directional terms herein, such as “top”, “bottom”, “left”, “right”, “up”, “down”, “upper”, “lower”, “inner”, “outer”, “medial”, “outer side”, “inner”, “external”, “proximal”, “distal”, etc., are used only to indicate relative directions and are not intended to otherwise limit the scope of the present disclosure or the claims. As used in the specification and the appended claims, the directional terms herein, such as “top”, “bottom”, “left”, “right”, “up”, “down”, “upper”, “lower”, “inner”, “outer”, “inner side”, “outer side”, “inner”, “external”, “proximal”, “distal”, etc., are used only to indicate relative directions and are not intended to otherwise limit the scope of the present disclosure or the claims.

[0120] Where possible, similar reference numerals have been used in the accompanying drawings. In addition, the alternative configurations of specific elements can each include a separate letter attached to the element number. Accordingly, additional letters can be used to specify an alternative design, structure, function, specific implementation and / or embodiment of an element or feature that does not contain additional letters. For example, element "80" can be embodied in an alternative configuration and is designated as "80a". Similarly, multiple instances of the sub-element of an element and / or a parent element can each include a separate letter attached to the element number. In each case, element markings can be used without the need to generally refer to all instances of an element or the additional letters of any one of the alternative elements. The element markings comprising additional letters can be used to refer to a specific instance of an element, or to distinguish the multiple uses of an element, or to attract the attention of the multiple uses of an element.

[0121] Various aspects of the devices, systems and methods of the present invention may be presented with reference to one or more exemplary embodiments. As used herein, the term "embodiment" means "serving as an example, instance or illustration" and should not necessarily be construed as being preferred or advantageous over other embodiments disclosed herein.

[0122] A number of different aspects of the apparatus and systems of the present invention may be presented by describing a plurality of components that are coupled, attached, and / or joined together. As used herein, the terms "coupled," "attached," "connected," and / or "engaged" are used to indicate a direct connection between two components, or an indirect connection to each other through an intervening or intermediate component, as appropriate. In contrast, when a component is referred to as being "directly coupled," "directly attached," "directly connected," and / or "directly linked" to another component, there are no intermediate elements.

[0123] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those of ordinary skill in the art to which the disclosure belongs. Although a variety of methods and materials similar or equivalent to those described herein can be used in the practice of the present disclosure, preferred materials and methods are described herein.

[0124] The specific implementation of the present disclosure extends at least to the pipette (e.g., multichannel pipette) for electroporation, and the electroporation system and / or its parts utilizing such pipette. The disclosed aspects and embodiments can be implemented to address the various shortcomings associated with at least some conventional pipettes and electroporation systems and / or technologies. The following discussion summarizes some example improvements and / or practical applications that can be provided by the disclosed embodiments. However, it should be understood that the following are merely examples, and the embodiments described herein are by no means limited to the example improvements discussed herein.

[0125] Some specific implementations of the present disclosure provide pipettes designed to simplify the pipetting operations required for sample processing and to reduce muscle strain associated with using a pipette to electroporate a sample for the pipette user. The unique design of the pipette described herein reduces muscle stress and / or fatigue in the user by reducing the force associated with manually performing the pipetting function to repeatedly process the sample. In addition, the pipette of the present disclosure is designed to utilize a pipette tip, wherein a clip-type connection between the pipette tip and the pipette is implemented. The use of a clip-type connection combined with pipette design improvements, as well as other components of the electroporation system, improves sample processing efficiency and reliability, as well as the ease of use of the electroporation system.

[0126] Some existing electroporation pipettes have a pipette tip with a conductive plunger component for carrying current during electroporation. However, such devices can exhibit high friction caused by interference between the plunger and the walls of the chamber (or lumen) of the pipette tip, which can affect the function of the pipette and / or cause delays during dispensing and / or aspiration.

[0127] At least some specific implementations of the present disclosure provide an electroporation pipette tip with a plunger, the plunger including a sealing component for reducing the contact area between the plunger and the chamber (or lumen) wall of the pipette tip. The sealing component can take various forms, such as a polymer sleeve and / or an O-ring. Such features can reduce the friction between the plunger and the chamber (or lumen) wall, thereby promoting improved pipetting function, making it less likely to degrade over time.

[0128] Conventional consumable pipette tips are usually connected to conventional electroporation pipettes via interference fit. In order to install conventional pipette tips to conventional electroporation pipettes, it is usually necessary for the user to apply significant downward force to press the electroporation pipette into the pipette tip, while maintaining the force on the pipette plunger trigger (e.g., actuator) so that the plunger holder of the electroporation pipette can clamp the plunger of the pipette tip. This may cause user fatigue and / or frustration. In addition, the conventional pipette tips of conventional electroporation pipettes often need to apply a large amount of force (e.g., about 60N) to the ejection button to allow the pipette tip to eject, which may further cause user fatigue and / or frustration.

[0129] At least some specific implementations of the present disclosure provide a pipette assembly that realizes a clip-type connection between a pipette tip and a pipette (e.g., one or more tabs of a pipette tip are clipped onto the holding platform of the pipette). Such features allow a two-part pipette tip attachment process, wherein the user presses the pipette into the pipette tip to facilitate the clip-type connection with the tip sleeve of the pipette tip, and subsequently or simultaneously actuates the actuator (e.g., trigger, button, etc.) by pressing the actuator so that the clamp jaws of the pipette reversibly grip / clamp the plunger of the pipette tip. This function can provide a more convenient pipette tip loading process for the user, thereby reducing muscle stress and / or fatigue. In various embodiments, the user can optionally actuate the first actuator (e.g., depress the plunger trigger), while the pipette is pressed into the pipette tip to perform tip loading. Similarly, a clip-on connection between a pipette tip and a pipette can allow for a two-part pipette tip ejection process, wherein a user first depresses a second actuator (e.g., an ejection button) to eject a tip sleeve (e.g., an outer portion of a pipette tip) without ejecting a plunger, and then depresses a first actuator of the pipette to release the plunger from the gripper jaws of the pipette tip. Such functionality can reduce peak ejection forces to facilitate tip separation / ejection, and can thereby reduce muscle stress and / or fatigue.

[0130] In many existing electroporation systems, the reservoir (e.g., buffer tube or other reservoir) holding the buffer solution is easily removed from the pipette docking assembly to prevent accidental removal of the reservoir from the assembly when the pipette tip is extracted from the reservoir of the assembly. This accidental removal may cause overflow and / or damage to the pipette tip. In some embodiments, the reservoir (e.g., buffer tube) is held by a pipette station guard assembled with the pipette station to protect the user from electric shock. On the contrary, conventional station guards are easily accidentally removed in the process of removing the pipette from the reservoir or even in the electroporation process, which presents a risk of electric shock.

[0131] At least some specific implementations of the present disclosure provide a pipette station guard that locks into a docking station (e.g., a "pipette station") via movement of the station guard in a locking direction that is different from a pipette removal direction for removing a pipette from a reservoir. The reservoir is inserted into an opening of the station guard and locked to the station guard. The reservoir can be released from the station guard by applying a force in a force direction different from the pipette removal direction for removing the pipette from the reservoir (e.g., on a latch member). Such features reduce or eliminate the incidence of accidental removal of a reservoir (e.g., a buffer tube) and / or a station guard from a pipette station during pipette removal, thereby reducing or avoiding spillage and / or pipette tip damage.

[0132] In many existing electroporation systems, the high voltage cable connecting the pipette station to the electrical pulse generator is integrally formed with the pipette station, and when a cable failure occurs (e.g., due to aging and / or degradation of the cable insulation), the entire pipette station typically needs to be replaced.

[0133] In contrast, the present disclosure provides a pulse generator having an external high voltage cable connection port (and, in at least some cases, a low voltage cable connection port) to enable the high voltage cable to be independent of the pulse generator, thereby enabling the replacement of the individual cable when a cable failure occurs (as opposed to having to replace the entire pulse generator in response to a cable failure). At least some specific implementations of the present disclosure may also provide a cable adapter that can be selectively mounted to the pulse generator, thereby facilitating improved cable management functions (e.g., consolidating cables during storage and / or managing cables connected to multiple pipette stations). In addition, at least some specific implementations of the present disclosure may provide a high voltage cable that is coupled to the low voltage cable via a braid and clamp assembly, which further facilitates convenient cable management.

[0134] In many existing electroporation systems, the pulse generator includes separate high-voltage and low-voltage power supplies for facilitating electroporation (via the high-voltage power supply) and other incidental functions, such as data transmission (via the low-voltage power supply). In the event of a failure in the isolation between the high-voltage circuit and the low-voltage circuit, a high-voltage discharge may occur by contact with a low-voltage component (e.g., a connection port such as a USB port, a LAN port, a Wi-Fi dongle port, etc.). At least some conventional electroporation systems include port covers on the low-voltage components to prevent harm to the user from the high-voltage discharge through the low-voltage components. However, conventional port covers typically require manual removal of the port cover to enable the external component to be connected to the port below, and typically require manual replacement of the port cover after the external component is disconnected from the port below. Users often forget to reconnect the port cover after the external component is disconnected, which may expose the user to the risk of injury and / or death due to high-voltage discharge through the low-voltage connection port.

[0135] At least some specific implementations of the present disclosure provide a low voltage port door system that is capable of automatically closing the port door after an external component is disconnected from a port associated with the port door. For example, each individual port door may include a biasing member that constantly biases the port door toward a closed position, thereby automatically forcing the port door into the closed position after an interfering object (e.g., a plug of an external component) is removed. Such functionality can reduce user exposure to risks associated with high voltage discharges through low voltage connection ports. The port door system may still require manual user action to open the port door for initial connection of an external component to the port. Such a port door system will comply with various safety compliance standards (e.g., IEC61010-1:2010 / AMD1:2016).

[0136] Many conventional electroporation systems include a pulse generator that uses a charging circuit to charge a large capacitor to a target voltage, and then uses a high-speed, high-voltage electronic switch to connect the capacitor to a pipette station to deliver a high-voltage pulse to the target cell (e.g., within a pipette chamber connected to a docking station). In at least some cases, such as when the load resistance is small (e.g., for larger pipette tip sizes), the voltage applied to the target cell may drop during the duration of the pulse. The drop in voltage can adversely affect the electroporation results.

[0137] At least some embodiments of the present disclosure provide a pulse generator controlled by a feedback loop, wherein a capacitor is charged to a voltage higher than a target voltage, and during discharge, the voltage is adjusted to produce a stable supply according to the target voltage (and / or pulse width and / or waveform settings). Such functionality can improve the consistency and predictability of electroporation results.

[0138] During high voltage electroporation (e.g., 500V to 2,500V), arcing may occur in response to bubbles and / or other contaminants in the chamber / lumen of the pipette tip. Arcing can lead to poor electroporation results and / or electroporation failure. Conventional electroporation systems fail to include a system for detecting arcing during electroporation. Therefore, in order to determine whether an arc has occurred, users typically rely on real-time visual monitoring of target cells to detect whether sparks are observed during electroporation. However, because such sparks occur in the millisecond time range, users often cannot detect visible sparks during electroporation. In addition, in some cases, arcing may occur in the absence of visible sparks (e.g., when the electroporation voltage is relatively low).

[0139] At least some specific implementations of the present disclosure include an arc detection module for automatically detecting a sudden drop in electroporation pulse current that indicates an arc. The arc detection module may include an amplifier, a bandpass filter, and a comparator to detect whether a falling current signal exists under various current distributions (e.g., for different types / sizes of pipette tips, for different buffer solutions, etc.). When an arc is detected, a notification or other indication that an arc has occurred may be provided to the user. Thus, the user can be aware of whether an arc has occurred during electroporation without relying on human monitoring during electroporation, which can help the user correctly interpret the electroporation results.

[0140] Now turn your attention to Figures 1 to 96 , which provide various supporting illustrations related to the disclosed embodiments as described in detail herein.

[0141] Electroporation system

[0142] Figure 1 Various example components of an electroporation system 100 that can be used to implement one or more disclosed embodiments are shown. For example, Figure 1 The electroporation system 100 can be configured to facilitate cell transfection by applying an electric current to target cells to introduce a payload into the target cells, so as to facilitate the production of genetically modified cells that can be used, for example, for cell therapy products. Figure 1 The electroporation system 100 is shown as including specific components, but in view of the present disclosure, it should be understood that the electroporation system 100 can include any number of additional and / or alternative components. In addition, in view of the present disclosure, it should be understood that the principles disclosed herein are not limited to Figure 1 The specific form and / or features of the electroporation system 100 or specific components thereof are shown.

[0143] Figure 1 The electroporation system 100 is shown to include a processor 102, a storage device 104, an input / output system 110 (I / O system 110), and a communication system 112. The processor 102 may include one or more sets of electronic circuits that include any number of logic units, registers, and / or control units to facilitate the execution of computer-readable instructions (e.g., instructions that form a computer program). Such computer-readable instructions may be stored in the storage device 104. The storage device 104 may include physical system memory and may be volatile, non-volatile, or some combination thereof. In addition, the storage device 104 may include a local storage device, a remote storage device (e.g., accessible via the communication system 112 or otherwise), or some combination thereof. Additional details related to a processor (e.g., processor 102), a computer storage medium (e.g., storage device 104), and other computer components will be provided below.

[0144] Processor 102 may be configured to execute instructions 106 stored in storage device 104 to perform certain actions and / or commands (e.g., voltage / current control, user interface presentation, receiving user input, component detection, etc.). These actions may rely, at least in part, on data 108 stored on storage device 104 in a volatile or non-volatile manner.

[0145] In some cases, the actions may rely at least in part on a communication system 112 for receiving data from a remote system 114, which may include, for example, a computing device, a sensor, and / or other. The communication system 112 may include any combination of software or hardware components that are operable to facilitate communication between components / devices on the system and / or with components / devices outside the system. For example, the communication system 112 may include a port, a bus, or other physical connection means for communicating with other devices / components. Additionally or alternatively, the communication system 112 may include systems / components that are operable to communicate wirelessly with external systems and / or devices via any suitable communication channel (such as, as non-limiting examples, Bluetooth, ultra-wideband, WLAN, infrared communication, and / or other).

[0146] also, Figure 1 The electroporation system 100 is shown to include or communicate with an I / O system 110. The I / O system 110 may include any type of input or output device, such as, but not limited to, a display, a touch screen, a mouse, a keyboard or button interface, a controller, etc., as non-limiting examples. For example, Figure 1 The electroporation system 100 is shown to include a user interface element implemented in the form of a graphical touch screen user interface on the pulse generator 120. The user interface element is configured to display information related to the operation of the electroporation system 100 and / or receive user input for facilitating control of the electroporation system 100 (e.g., to select parameters for an electroporation procedure, to initiate, monitor, and / or to terminate an electroporation procedure).

[0147] The electroporation system 100 includes various physical components that can be used to facilitate the electroporation operation. For example, Figure 1The electroporation system 100 is shown to include a pulse generator 120 configured to provide electrical pulses to other components of the electroporation system 100. The pulse generator 120 can provide electrical pulses via a cable 122, which can selectively connect the pulse generator 120 to one or more other components of the electroporation system 100. For example, the cable 122 can connect the pulse generator 120 to a pipette docking assembly 121 to supply current to target cells residing in a pipette tip of a pipette 130 connected to the pipette docking assembly 121. The pipette docking assembly 121 can include a pipette station 124 (to which the cable 122 can be connected), a pipette station guard 126, and a reservoir (e.g., a buffer tube 128 that receives the pipette 130). Additional aspects of the components of the electroporation system 100 will be described in more detail below.

[0148] Pipette Tips

[0149] FIG. 2A to FIG. 7B and Figures 83 to 96 Various aspects of a pipette tip for use in an electroporation system of the present disclosure are shown. Such a pipette tip can be connected to a pipette (e.g., pipette 130) and can hold a sample containing cells and a payload to facilitate electroporation. Figure 2A An example of a 10 μL pipette tip 202 (eg, sized to hold a sample volume of approximately 10 μL) is depicted, and Figure 2B A 100 μL pipette tip 204 is depicted (eg, sized to hold a sample volume of approximately 100 μL). Figure 2A and Figure 2BOnly sizes of 10 μL and 100 μL are shown in the figures, respectively, but other sizes are also within the scope of the present disclosure, including any pipette tips whose sizes are designed to accommodate the following sample volumes: 0.1 μL to 500 μL, 0.1 μL to 450 μL, 0.1 μL to 400 μL, 0.1 μL to 350 μL, 0.1 μL to 300 μL, 0.1 μL to 250 μL, 0.1 μL to 200 μL, 0.1 μL to 150 μL, 0.1 μL to 100 μL, 0.1 μL to 90 μL, 0.1 μL to 80 μL, 0.1 μL to 70μL, 0.1μL to 60μL, 0.1μL to 50μL, 0.1μL to 40μL, 0.1μL to 30μL, 0.1μL to 20μL, 0.1μL to 10μL, 0.0.1μL to 5μL, 0.10μL to 250μL, 0.10μL to 100μL, 0.10μL to 50μL, 1μL to 500μL, 1μL to 450μL, 1μL to 400μL, 1μL to 350μL, 1μL to 300μL, 1μL to 250μL, 1μL to 200μL, 1μL to 1μL to 100μL, 1μL to 90μL, 1μL to 80μL, 1μL to 70μL, 1μL to 60μL, 1μL to 50μL, 1μL to 40μL, 1μL to 30μL, 1μL to 20μL, 1μL to 10μL, 1μL to 10μL, 10μL to 500μL, 10μL to 450μL, 10μL to 400μL, 10μL to 350μL, 10μL to 300μL, 10μL to 250μL, 10μL to 200μL, 10μL to 150μL, 10 Sample volumes between μL to 100μL, 10μL to 90μL, 10μL to 80μL, 10μL to 70μL, 10μL to 60μL, 10μL to 50μL, 10μL to 40μL, 10μL to 30μL, 10μL to 20μL, including any increments therebetween, including 0.2μL, 0.5μL, 1μL, 5μL, 10μL, 25μL, 50μL, 75μL, 100μL, 150μL, 200μL, 250μL, 300μL, 350μL, 400μL, 450μL, etc.

[0150] Figure 3B 100 μL pipette tip 204 is shown to include a plunger 302 configured to be at least partially disposed within a lumen 308 of a tip barrel 304 ( Figure 2BThe plunger 302 is shown fully inserted into the lumen 308 defined by the tip sleeve 304. The plunger 302 is configured to translate along the length of the lumen 308 to facilitate pipetting functions (e.g., aspiration and / or dispensing). For example, the distal open end 306 of the lumen 308 can be positioned within a container containing a liquid sample including cells and a payload (e.g., nucleic acids, proteins, etc.), and the plunger 302 can be withdrawn (e.g., proximally) from the distal open end 306 of the lumen 308 to draw (e.g., aspirate) the cells and payload into the lumen 308. The 100 μL pipette tip 204 can then be connected to other components of the electroporation system (e.g., Figure 1 A pipette docking assembly) is used to electroporate the cells to introduce the payload into the cells.

[0151] At least a portion of the plunger 302 may include a conductive material to enable electrical pulses to reach and / or travel through the contents of the lumen 308. For example, the plunger 302 may be coated with, formed of, or otherwise include gold (e.g., gold plating), diamond-like carbon, conductive plastic, and / or any other conductive medical grade material (e.g., a material that is inert to mammalian cells).

[0152] Figure 3A It is shown that the 10 μL pipette tip 202 may include a plunger 310 and a tip sleeve 312 having an internal cavity 316 (with a distal open end 314 ) similar to the plunger 302 and internal cavity 308 of the 100 μL pipette tip 204 .

[0153] For conventional pipette tips, a seal is formed between the plunger and the inner cavity wall defining the inner cavity by a metal ring on the plunger that docks with the inner cavity wall. The amount of frictional force exhibited between the metal ring and the inner cavity wall may affect the push / pull force required to operate the pipette. The amount of frictional force exhibited between the metal ring and the inner cavity wall may be affected by the amount of interference between the metal ring and the inner cavity wall. By way of example, for a 10 μL tip, interference within the range of 0 μm to 30 μm may produce a push / pull force within the range of 0 N to 6 N to operate the pipette. For larger tips, such as 10 μL tips, interference within the range of 0 μm to 10 μm may produce a push / pull force within the range of 0 N to 6 N to operate the pipette. It may be difficult to consistently and reliably achieve interference in the 0 μm to 10 μm range in production, which may result in pipette tips (especially larger pipette tips) having excessive interference between the metal ring and the inner cavity wall, resulting in excessive push / pull force (e.g., exceeding 6N) required to operate the pipette.

[0154] Therefore, at least some pipette tips of the present disclosure may implement alternative sealing components for creating a seal between the lumen wall and the plunger. In some embodiments, this is beneficial for larger sized pipette tips (e.g., 50 μL or 100 μL pipette tips or larger pipette tips).

[0155] FIG. 4A to FIG. 4B (To decompose the configuration( Figure 4A ) and assembly configuration ( Figure 4B ) Both) show an example plunger 402 of a 100 μL pipette tip. FIG. 4A to FIG. 4B In the embodiment of the present invention, the plunger 402 includes an engagement segment 404 and an inner cavity segment 406. The engagement segment 404 is configured to be operably engaged with the gripper jaws of the pipette, which are configured to grasp and release the engagement segment 404 to hold and release the engagement segment 404, respectively, as will be described in more detail below. The inner cavity segment 406 is configured to be positioned within the inner cavity of the pipette tip sleeve of the pipette tip and translate along the length of the inner cavity.

[0156] like FIG. 4A to FIG. 4B As shown, the lumen section includes a sealing member 410 that creates a seal between the plunger 402 and the lumen wall of the lumen in which the plunger is positioned. FIG5 shows a cross-sectional view of the lumen section 406 of the plunger 402 positioned within the lumen of a pipette tip sleeve 506. Figure 5B An interference region is depicted between the sealing member 410 and the inner wall 502 defining the lumen of the tip sleeve 506. Advantageously, the interference region does not extend along the entire length of the lumen section 406 of the plunger 402 positioned within the lumen, thereby helping to reduce friction between the plunger 402 and the lumen wall.

[0157] exist FIG. 4A to FIG. 4B and FIG. 5A to FIG. 5B In the example of FIG. 4 , the sealing component 410 is implemented as a polymer sleeve (other forms are also possible, such as Figure 6 and FIG. 7A to FIG. 7B The sealing member 410 may include various types of materials, such as polytetrafluoroethylene (PTFE), other Teflon materials, and / or other flexible and biocompatible materials.

[0158] The sealing member 410 may be attached to the lumen section 406 of the plunger 402 in a variety of ways. FIG. 4A to FIG. 4B and FIG. 5A to FIG. 5B In the example of , the lumen segment includes a front pin 412 and a shaft segment 414. The front pin 412 is configured to be connected to the shaft segment 414, such as by inserting a portion of the front pin 412 into a retaining hole 416 of the shaft segment 414. The front pin 412 can also secure the sealing component 410 to the shaft segment 414, such as by inserting the front pin 412 through an opening in the sealing component 410 before the front pin 412 enters the retaining hole 416 of the shaft segment 414.

[0159] In some cases, when sealing component 410 is secured to lumen segment 406, a space is formed between at least a portion of sealing component 410 and at least a portion of lumen segment 406. This can facilitate flexibility of sealing component 410 for creating a seal between lumen segment 406 and a wall defining the lumen (e.g., reducing friction therebetween while still maintaining a seal). Figure 5B A space 504 formed between the sealing member 410 and the front pin 412 when the front pin 412 is inserted through the sealing member 410 is shown.

[0160] In view of this disclosure, it should be appreciated that other methods for securing the sealing component 410 to the lumen segment 406 (eg, adhesives, mechanical fit, threaded connections, etc.) may be implemented in accordance with the present disclosure.

[0161] As mentioned above, the sealing member of the plunger may take a variety of forms. Figure 6 and FIG. 7A to FIG. 7B An alternative form of sealing member is shown. Figure 6 Plunger 602 is shown in which the sealing component is implemented as an O-ring 604 , which may be coated (eg, with an inert lubricating material). An inner lumen section 606 of plunger 602 includes a circumferential recess 608 configured to receive O-ring 604 . FIG. 7A to FIG. 7B An O-ring 604 is shown mating with an inner wall 702 defining a lumen of a tip sleeve 704 to form a seal between the lumen segment 606 and the inner wall 702. In some cases, an O-ring design may require greater force than a polymer sleeve design to promote a seal between the plunger and the pipette lumen (e.g., due to the lack of internal space in an O-ring design).

[0162] Pipettes and Pipette Components

[0163] Fig. 8A A pipette 820 is shown for use in the systems of the present disclosure, which may be distally attached to a pipette tip, such as Figure 8B Pipette tips shown. Figure 76 to Figure 82Various views of a pipette 820 with attached pipette tips are shown for use in the systems of the present disclosure. It should be understood that while the present disclosure shows embodiments of a pipette configured to couple with a single pipette tip, the pipettes of the present disclosure can be configured to couple with 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or more pipette tips while maintaining the same functionality (e.g., using 2 proximally positioned actuators to control pipetting functions and tip attachment) by expanding the pipette design to include multiple channels with the same or similar structural features as disclosed herein for performing pipetting functions and attaching / detaching pipette tips, including clamping tip connections, gripping / releasing plungers, and controlling the movement of plungers to perform pipetting functions. Thus, the pipette of the present disclosure may be configured to reversibly couple a plurality of pipette tips whose functions are controlled by two actuators proximally disposed on the pipette.

[0164] In various embodiments, the pipette includes a proximal segment having a handle and a distal segment configured to be reversibly attached to a pipette tip. The pipette also includes a first actuator and a second actuator that are operable to control the functions of the pipette. In some embodiments, the first actuator is disposed in the proximal segment and, when actuated, is operable to control: i) the pipetting functions of the pipette (aspiration and dispensing of a sample); and ii) the gripping and release of a plunger disposed within the lumen of the pipette tip (see, e.g., 9A to 10B ). In a related embodiment, the second actuator is disposed in the proximal section and, when actuated, is operable to separate the tip sleeve of the pipette tip from the distal section of the pipette. In various embodiments, both the first actuator and the second actuator are actuated by depressing and / or releasing the actuator. It should be understood that each of the first actuator and the second actuator is oriented so that operation / control of the respective function of each actuator can be controlled by the user's thumb gripping the handle of the pipette. The pipette also includes a pipette electrode that contacts an electrode disposed on the docking station when the pipette is docked in the pipette docking assembly (see, e.g. Figure 1 ).

[0165] As further discussed and illustrated herein, in embodiments, the first actuator has a first undepressed position and a second partially depressed position, in which the actuator moves distally relative to the handle. When the pipette tip is attached and the plunger is grasped by the jaws of the holder, transitioning the first actuator from the first undepressed position to the second partially depressed position results in dispensing from the pipette tip by translation of the plunger in the lumen of the pipette tip, and transitioning the first actuator from the second partially depressed position to the first undepressed position results in aspiration into the pipette tip by translation of the plunger in the lumen of the pipette tip.

[0166] As further discussed herein, the first actuator has a third fully depressed position, in which the first actuator is pushed distally relative to the handle past the second part depressed position. When the plunger is gripped by the clamp jaws, the first actuator is transformed from the second part depressed position to the third fully depressed position causing the opening of the clamp jaws and the release of the plunger. The first actuator is transformed from the third fully depressed position to the second part depressed position causing the gripping of the plunger. In the process in which the pipette tip is attached to the pipette, the first actuator is transformed to the third fully depressed position, the engagement section of the plunger is directed to the distal opening of the clamp jaws, and then the first actuator is transformed to the second part depressed position to realize the closure of the clamp jaws and the gripping of the engagement section, so that the suction and distribution functions are controlled by the first actuator via the translation of the inner cavity of the plunger and the pipette tip in the first position and the second position. During separation of the pipette tip from the pipette, the first actuator transitions to the third, fully depressed position to loosen and release the engagement segment of the plunger, and if the tip sleeve has not yet been separated by actuation of the second actuator, the first actuator may remain in the third position until separation of the tip sleeve is complete, as further discussed herein.

[0167] As further discussed herein with reference to specific drawings, in various embodiments, the pipette includes a clamp mechanism operably coupled to the first actuator, the clamp mechanism having a clamp jaw and a clamp sleeve disposed around the clamp jaw. In some embodiments, the operation of the clamp mechanism is controlled by the first actuator, and when the first actuator is switched between the second partially depressed position and the third fully depressed position, the closed configuration (for gripping) and the open configuration (for releasing) are switched. The clamp jaw includes a jaw opening for receiving the engagement portion of the plunger, and when the first actuator is in the first undepressed position and the second partially depressed position via relative positioning with the clamp sleeve, the clamp jaw can be operated to grip the engagement segment of the plunger. For example, the clamping sleeve is positioned around the clamp jaws and is configured to apply an inward force to the clamp jaws so that the clamp jaws apply a compressive force to the engagement segment of the plunger to retain the engagement segment of the plunger within the jaw opening when the first actuator is in the first depressed position or the second partially depressed position.

[0168] In embodiments, the actuation of the second actuator causes the separation of the tip sleeve of the pipette tip, separated from the operation of the first actuator, to control the engagement and movement of the plunger. In some embodiments, the second actuator has a first unpressed position and a second depressed position, in which the actuator moves distally toward the distal section of the pipette. In other embodiments, the tip ejects a sleeve operably connected to the second actuator and disposed adjacent to the tip interface of the pipette, so that when the second actuator is actuated by transitioning the second actuator from the first unpressed position to the second depressed position and causing the tip sleeve to shift and separate from the distal section of the pipette, the tip ejects a sleeve relative to the tip interface and moves distally.

[0169] As discussed throughout, pipette tips may be selectively attached to the pipettes of the present disclosure. Figure 8B A pipette tip 802 is shown, which may correspond to any of the pipette tips discussed herein. Fig. 8A A pipette 820 is shown. FIG. 8A to FIG. 8BIn the example of the embodiment of the present invention, the pipette tip 802 includes an attachment interface 806 adjacent to its inner cavity 804. The attachment interface 806 includes tabs 808 that are configured to engage with corresponding attachment features of the pipette 820 (any number of tabs may be utilized). In an embodiment, the tabs are angled toward the inner cavity of the pipette tip 802. The corresponding attachment features of the pipette 820 are arranged on the distal segment 822 of the pipette 820 as a tip interface 830 including a holding platform 832. The distal segment 822 of the pipette may include a segment of the pipette 820 opposite to the proximal segment 824 of the pipette 820, which includes an actuator for manually controlling to operate the pipette 820 (e.g., a first actuator 828 and a second actuator 826 that control the pipetting function and plunger engagement and tip sleeve separation / ejection, respectively).

[0170] Fig.9A Example attachment features (e.g., a tip interface) of the pipette 820 and corresponding attachment features (e.g., an attachment interface) of the pipette tip 802 are shown. Fig.9A The corresponding attachment features are shown separated, for example, a pipette tip and a pipette are separated from each other. Fig. 9B A corresponding attachment feature of the coupling is shown, for example, attached to a pipette tip sleeve of a pipette. Specifically, Fig.9A The holding platform 902 of the tip interface 830 of the distal segment 822 of the pipette is shown. The holding platform 902 resides on top of the curved surface 904 and within the recess 906 of the distal segment 822. This allows the tab 808 of the pipette tip 802 to advance into engagement with the holding platform 902. For example, when the distal segment 822 of the pipette 820 is pressed into the attachment interface 806 of the pipette tip 802, the tab 808 of the attachment interface can advance on the angled surface 904 and expand until it reaches the recess 906, at which point the tab 808 can retract inwardly toward the recess 906 and engage with the holding platform 902.

[0171] In some cases, after the tab 808 reaches the recess 906 and retracts therein, the biasing member of the pipette 820 can be operated to bias the tab 808 into engagement with the retention platform 902 . Fig. 9C A biasing member 920 of the distal section 822 of the pipette 820 is shown, the biasing member comprising a spring 922 and a biasing platform 924 . Fig. 9C The attachment interface 806 of the pipette tip 802 is shown arranged to be ready for connection to the distal section 822 of the pipette 820. When the attachment interface 806 is advanced on the distal section 822 of the pipette 820 (according to Fig. 9C808 has reached the recess 906), the spring 922 forces the biasing platform 924 to abut against the attachment interface 806 to force its tab 808 to engage with the retaining platform (as shown in the arrows). Fig.9D shown and indicated by its thick arrow).

[0172] Fig.9D The plunger 930 of the pipette tip 802 is shown. The plunger 930 (particularly the engagement section 932 of the plunger 930) can become reversibly clamped by a clamp jaw, the proximal region of which is surrounded by a clamp sleeve, the clamp jaw and the clamp sleeve forming a clamp mechanism being disposed in the distal section 822 of the pipette 820. Fig.9E A gripper mechanism 940 is shown that includes a gripper jaw 942 and a clamping sleeve 946. The gripper jaw 942 includes a distally disposed jaw opening 944 for receiving the engagement segment 932 of the plunger 930 so that it can be held (e.g., grasped) by the gripper jaw 942. In some implementations, the gripper jaw 942 includes an at least partially flexible material to enable the engagement segment 932 to be inserted therein. The bevel or bend at the top portion of the engagement segment 932 of the plunger 930 can improve the ease of inserting the engagement segment into the gripper jaw 942.

[0173] The gripper mechanism 940 is operably connected to the first actuator 828 and can be actuated by the first actuator to facilitate advancement of the gripper jaws 942 into engagement with the engagement portion 932 of the plunger 930. For example, the gripper mechanism can be actuated by the first actuator 828 of the pipette 820 (see Fig. 8A) operation to actuate. In an embodiment, during actuation of the first actuator (e.g., from a first undepressed position to a second partially depressed position), a holder sleeve 946 in line with the holder jaws 942 may be pushed distally a distance. Subsequently, during further actuation, only the holder jaws 942 are advanced to open the holder jaws 942 to receive the engagement segment 932 (e.g., from a second partially depressed position to a third fully depressed position), which is coordinated with the tab 808 of the attachment interface 806 of the pipette tip 802 being simultaneously advanced to engage with the holding platform 902 (e.g., by depressing the first actuator 828 while pressing the pipette 820 and the pipette tip 802 into each other). Alternatively, during actuation of the first actuator (e.g., from a first, undepressed position to a second, partially depressed position), the clamp sleeve 946 in line with the clamp jaw 942 can be pushed distally forward a distance and then, during further actuation, only the clamp jaw 942 is advanced to open the clamp jaw 942 to receive the engagement segment 932 (e.g., from the second, partially depressed position to a third, fully depressed position), and the tab is pushed into engagement with the retaining platform 902 (e.g., by first pressing the tab 808 into engagement with the retaining platform 902 and then depressing the first actuator 828 and translating the first actuator from the first, undepressed position to the third, fully depressed position) at different times than the actuation of the first actuator and the engagement of the clamp jaw 942 with the engagement segment 932.

[0174] The plunger mechanism 940 also includes a clamping sleeve 946 positioned around the clamp jaws 942, such as Fig.9E The clamp sleeve 946 is configured to apply an inward force to the clamp jaw 942 so that the clamp jaw 942 applies an inward compressive force on the engagement section 932 to grasp and hold the engagement section 932 of the plunger 930 within the clamp jaw 942. In some specific implementations, the clamp sleeve 946 includes a material that is at least partially more rigid than the material of the clamp jaw 942.

[0175] After clamping the plunger 930, the clamp mechanism 940 can be moved by the first actuator 828 of the pipette 820 (see Fig. 8A ) operation to facilitate the pipetting function. Fig. 10A and Fig. 10B The gripper mechanism 940 is shown in the aspiration direction toward the proximal section of the pipette (see Fig. 10A , as indicated by the arrow therein, for example caused by movement of the first actuator from the second partially depressed position toward the first undepressed position) and in a dispensing direction toward the distal section of the pipette (see Fig. 10B, as indicated by the arrow therein, e.g., caused by movement of the first actuator from the second partially depressed position toward the first undepressed position. When the plunger 930 is clamped by the clamp jaws 942 of the clamp mechanism 940, actuation of the clamp mechanism 940 causes the lumen section 1002 of the plunger 930 to translate within the lumen 1004 of the pipette tip 802.

[0176] In some embodiments, after the pipette tip 802 is engaged with the pipette 820 (e.g., with the tab 808 engaged with the retention platform 902 and the plunger 930 engaged with the clamp mechanism 940), the pipette tip 802 can be selectively ejected from the pipette 820 via a two-step process, which can reduce the peak force required to promote disengagement (compared to a single-action process for promoting disengagement). As a non-limiting example, while a single-action ejection process may require a peak force of approximately 60N, a multi-step ejection process as currently disclosed may require a peak force of approximately 40N. As described above, the pipette 820 includes a second actuator 826 (see Fig. 8A ), the second actuator can be used to disengage the attachment interface 806 from the distal section 822 of the pipette 820 (without disengaging the clamp mechanism 940 from the plunger 930).

[0177] For example, the second actuator 826, when pressed, can cause the tip to be ejected from the sleeve 950 (see Fig.9D ), so that the tip ejection sleeve 950 is advanced (e.g., downward) from the inner side of the tab 808 toward the tab 808. During the advancement, the tip ejection sleeve 950 can press on the inner side of the tab 808 to bend the tab 808 outward and disengage the tab 808 from the holding platform 902. The biasing member 920 can then press the attachment interface 806 of the pipette tip 802 downward from the pipette 820, thereby ejecting the attachment interface 806 of the pipette tip 802 (and the inner cavity attached thereto) from the pipette.

[0178] Fig.11A Schematic diagram of actuating (e.g., depressing) the second actuator 826 of the pipette 820 to cause the attachment interface (e.g., attachment interface 806) and the inner cavity (e.g., inner cavity 1004) of the pipette tip (e.g., pipette tip 802) to be ejected from the pipette 820. Fig.11AIn the example of , the second actuator 826 is configured to traverse the blank travel distance when depressed to move the second actuator from a first undepressed position to a second depressed position (e.g., a distance of about 6 mm) before the attachment interface 806 and the inner cavity of the pipette tip are disengaged from the pipette 820 by the tip ejection sleeve (this occurs by applying a force of about 20N to 30N to press the ejection button 826 through a final distance of about 2.5mm). Such functionality can prevent the attachment interface and the inner cavity of the pipette tip from being accidentally ejected from the pipette 820 (other blank travel and ejection distances can be used).

[0179] Fig. 11B is a schematic diagram of a pipette with an attachment interface and an inner cavity of a pipette tip ejected therefrom. In an embodiment, a first actuator is actuated to release the plunger from a clamping mechanism before actuating a second actuator to disengage the attachment interface. In some embodiments, the second actuator is first actuated to disengage the attachment interface, and then the first actuator is actuated to release the plunger from the clamping mechanism. Fig. 11B As depicted, depressing the first actuator 828 after the attachment interface 806 is disconnected from the pipette 820 causes the engagement segment of the plunger 930 to be ejected from the gripper mechanism 940 of the pipette. For example, depressing the first actuator 828 after the attachment interface 806 is disconnected from the pipette 820 can advance the gripper jaws 942 out of the gripper sleeve 946, thereby releasing the inward force previously applied by the gripper jaws 942 to the engagement segment 932 of the plunger 930 and ejecting the plunger 930 from the gripper mechanism 940. Fig. 11B In the illustrated embodiment, example forces associated with a plunger spring of approximately 12.32 N and a retainer spring of approximately 28.20 N may be overcome by depressing the first actuator 828 to facilitate plunger ejection (eg, resulting in a total force of approximately 40.52 N experienced by the user).

[0180] In various aspects, the present disclosure provides a pipette assembly including a pipette reversibly attached to a pipette tip. In some embodiments, the pipette of the assembly includes a proximal segment with a handle, a distal segment with a tip interface, and a first actuator disposed in the proximal segment. The pipette tip reversibly attached to the pipette of the assembly includes: a tip sleeve defining an inner cavity extending from the proximal end of the pipette tip to the distal end of the pipette tip; a plunger at least partially disposed in the inner cavity; and an attachment interface disposed at the proximal end of the pipette tip. In various embodiments, the plunger is reversibly operably coupled to the first actuator, and when operably coupled, performs a pipetting function when the first actuator is actuated by translating along the inner cavity. In addition, the tip sleeve is reversibly attached to the distal segment via one or more tabs of the attachment interface that engage with the retention platform of the distal segment. In an embodiment, the pipette of the assembly further comprises a pipette electrode disposed in the distal segment and electrically coupled to the plunger when the plunger is operably coupled to the first actuator.

[0181] In some embodiments, the pipette of the assembly includes a proximal section with a handle, a distal section with a tip interface, a first actuator disposed in the proximal section, and a clamp mechanism with a clamp jaw disposed in the distal section. The pipette tip reversibly attached to the pipette of the assembly includes: a tip sleeve defining an inner cavity extending from the proximal end of the pipette tip to the distal end of the pipette tip; and a plunger at least partially disposed in the inner cavity. In various embodiments, the plunger is reversibly operably coupled to the first actuator via the clamp jaw, and when operably coupled, performs a pipetting function when the first actuator is actuated by translating along the inner cavity. In an embodiment, the pipette of the assembly also includes a pipette electrode, which is disposed in the distal section and electrically coupled to the plunger when the plunger is operably coupled to the clamp jaw (e.g., grasped by the clamp jaw).

[0182] Pipette docking assembly

[0183] Fig.12 13 illustrate aspects of an example pipette station guard for an electroporation system (eg, electroporation system 100 ).

[0184] Fig.12 A pipette station guard 1202 is shown. The pipette station guard 1202 can be attached to the pipette station to protect the user from potential electric shock. The pipette station guard 1202 can additionally include a reservoir opening 1204 (e.g., a buffer tube opening) for receiving a reservoir (e.g., a buffer tube) that can receive a pipette (e.g., pipette 820) and / or a component connected thereto (e.g., a pipette tip).

[0185] To facilitate connection to the pipette station, the pipette station guard 1202 may include various connection elements, such as one or more locking hooks configured to engage with one or more corresponding hook fasteners of the pipette station. Such locking hooks may take various forms. For example, Fig.12 The example of the pipette station guard 1202 is shown as including one or more pivoting hooks 1206 that are configured to rotate into engagement with one or more hook fasteners (eg, pivoting hook fasteners) of the pipette station (see FIG. 13 ). Fig.12 The example also shows the pipette station guard 1202 as including one or more flexible hooks 1208 that are configured to be advanced into engagement with one or more hook fasteners of the pipette station (see FIG. 13 ).

[0186] Fig.12 The pipette station guard 1202 is also shown to further include one or more finger guides 1210 indicating one or more depressible surfaces of the pipette station guard 1202. The depressible surfaces of the pipette station guard 1202 are depressible to cause movement of the one or more flexible hooks 1208 to facilitate engagement / disengagement of the one or more flexible hooks 1208 with their corresponding hook fasteners of the pipette station (see FIG. 13 ).

[0187] exist Fig.12 In the example of , the pipette station guard 1202 includes a pair of pivoting hooks 1206 and a pair of flexible hooks 1208 arranged on the rear surface of the pipette station guard 1202. The pair of pivoting hooks 1206 are arranged on the top portion of the rear surface, and the pair of pivoting hooks are arranged on the bottom portion of the rear surface. Fig.12 Also depicted is a finger guide 1210 disposed on a bottom portion of a side surface of the pipette station guard 1202 (an opposing finger guide is disposed on a bottom portion of an opposing side surface of the pipette station guard 1202).

[0188] FIG. 13 shows the pipette station guard 1202 moved into engagement with the pipette station 1302 . Fig.13A The pivoting hook 1206 of the station guard 1202 is shown initially inserted into the corresponding pivoting hook catch 1304 of the pipette station 1302. After the pivoting hook 1206 is inserted, the station guard 1202 is rotated toward the pipette station 1302 (by Fig.13A ), until the flexible hook 1208 reaches the corresponding flexible hook fastener 1306 and interlocks therewith. Fig.13AAlso shown are finger guides 1210 of the pipette station guard 1202 that can be pressed to move the flexible hooks 1208 inwardly toward each other, thereby enabling the flexible hooks 1208 to easily enter openings associated with the flexible hook fasteners 1306. After the flexible hooks 1208 enter the openings, pressure can be released from the finger guides 1210 to allow the flexible hooks 1208 to move outwardly so that the flexible hooks 1208 interlock with the flexible hook fasteners 1306. Fig. 13B The pipette station guard 1202 is shown fully installed to the pipette station 1302.

[0189] In some embodiments, to remove the pipette station guard 1202 from the pipette station 1302 , the user may depress the finger guides 1210 to allow the flexible hooks to disengage from the flexible hook clasps 1306 and withdraw / rotate the pipette station guard 1202 to disengage from the pipette station 1302 .

[0190] As described above, the pipette station guard 1202 may include a reservoir opening 1204 for receiving a reservoir. Fig.14 An example reservoir 1402 (eg, a buffer tube) is shown that may be inserted into the reservoir opening 1204 of the pipette station guard 1202 (eg, when the pipette station guard 1202 is installed to the pipette station 1302). Fig.14 The reservoir 1402 includes a latch member 1404 that is configured to engage with a corresponding latch catch 1220 (see FIG. 1402 ) of the pipette station guard 1202 when the reservoir 1402 is inserted into the reservoir opening 1204 of the pipette station guard 1202. Fig.12 The latch member 1404 of the reservoir 1402 can enable the reservoir 1402 to be connected to the pipette station guard 1202 in a manner that prevents the reservoir 1402 from accidentally disconnecting from the pipette station guard 1202 during use (e.g., when pipette components are withdrawn from the reservoir 1402, thereby mitigating potential spillage and / or damage to the pipette components).

[0191] In some cases, the shape of the reservoir 1402 itself can help mitigate the risk of damaging the pipette tip. For example, the reservoir 1402 can include a lower section 1406 and an upper section 1408, the pipette tip being configured to reside in the lower section, and the distal section of the pipette being configured to reside in the upper section. The shape of the upper section 1408 can force the entry and extraction of the pipette in a manner that is longitudinally aligned with the reservoir 1402, thereby forcing the pipette tip to align with the lower section 1406 and prevent damage to it during insertion and / or extraction.

[0192] exist Fig.14 In the example of FIG. 14 , the latch member 1404 is arranged outside the reservoir 1402 . Fig.14It is also shown that the latch members 1404 may include corresponding finger tips 1410 for guiding the positioning of the user's fingers to apply force to one or more latch members 1404. Applying force to the latch members 1404 from the positioning of the finger tips 1410 may cause the latch members 1404 to bend / deform, thereby enabling them to engage or disengage with corresponding latch catches 1220 of the pipette station guard 1202. The finger tips 1410 may also guide the placement of the user's fingers when handling the reservoir so as to reduce the risk of human fingers contacting the pipette components during insertion and / or withdrawal of the pipette components.

[0193] exist Fig.14 In the example of , the latch member 1404 extends from a flange 1412 of the reservoir 1402, which extends around a pipette opening 1414 of the buffer tube 1402 (for receiving a pipette component). The flange 1412 can be at least partially tilted inwardly toward the pipette opening 1414 to guide any fluid that falls from the pipette component into the pipette opening. The flange 1412 can also be operated to provide space between the finger prompt 1410 and the pipette opening 1414, thereby helping to mitigate the risk of pipette component contamination (e.g., during insertion and / or extraction of the pipette component through the pipette opening 1414).

[0194] Fig.14 It is also shown that the reservoir 1402 may include a volume indicator 1416 to allow a user to determine when the reservoir 1402 contains a sufficient amount of buffer solution to perform electroporation. Fig.14 The reservoir 1402 also includes an electrode 1418 that can contact a corresponding electrode of the pipette station (or pipette station guard) to allow the electrical pulses from the pulse generator to reach the buffer solution within the reservoir 1402 (and the target cells within the pipette tip positioned within the buffer solution). In some cases, the reservoir 1402 includes one or more hooks or other retaining members for retaining pipette components (e.g., a pipette and / or a pipette tip) therein.

[0195] Fig.15A and Fig. 15B The reservoir 1402 is shown being inserted into the pipette station guard 1202 mounted to the pipette station 1302. As previously described, the latch members 1404 of the reservoir 1402 interlock with the corresponding latch catches 1220 of the pipette station guard 1202. Fig.16 A cross-sectional view of a reservoir 1402 positioned within a reservoir opening 1204 of a pipette station guard 1202 is shown, wherein a latch member 1404 of the reservoir 1402 interlocks with a corresponding latch catch 1220 of the pipette station guard 1202. Fig.16It can be readily seen that the direction of force applied to disengage the latch member 1404 from the corresponding latch catch 1220 (inward from the finger tip 1410) is different from the pipette withdrawal direction (inward from the finger tip 1410) used to withdraw the pipette component from the pipette opening 1414 of the reservoir 1402. Fig.16 ). As described above, this can mitigate the risk of the reservoir 1402 being accidentally withdrawn from the reservoir opening 1204 of the pipette station guard 1202 when withdrawing pipette components from the reservoir 1402.

[0196] Fig.17 A safety interlock feature 1702 of the pipette station 1302 is shown that is configured to engage / interact with a portion of the reservoir 1402 (e.g., a bottom portion thereof) when (i) the pipette station guard 1202 is connected to the pipette station 1302 and (ii) the latch member 1404 of the reservoir 1402 is engaged with the corresponding latch catch 1220 of the pipette station guard 1202. Fig.17 In the example of FIG. 1 , the safety interlock feature 1702 is implemented as an upward protrusion that prevents the reservoir 1402 from passing through the protrusion (e.g., in a direction for withdrawing the pipette station guard 1202). Thus, with the reservoir 1402 connected to the pipette station guard 1202, the safety interlock feature 1702 can prevent the pipette station guard 1202 from being accidentally removed from the pipette station 1302.

[0197] The height of the reservoir 1402 (which interfaces with the vertical wall of the pipette station 1302, such as Fig.17 The pipette station guard 1202 may further help prevent accidental removal of the pipette station guard 1202 from the pipette station 1302 (eg, by preventing rotation necessary to remove the pipette station guard 1202 from the pipette station 1302).

[0198] Fig.18A and Fig.18B It is shown that when the pipette station guard 1202 is connected to the pipette station 1302 and the reservoir 1402 is connected to the pipette station guard 1202 as described above, the pipette 820 and associated pipette tip 802 can be inserted into the reservoir 1402 to facilitate electroporation. Fig.19 As shown, as described above, the direction for withdrawing or inserting the pipette 820 (and / or associated pipette tips) from the reservoir 1402 is different from the direction in which the locking hooks of the pipette station guard 1202 are configured to retract from engagement with the hook fasteners of the pipette station 1302. Similarly, Fig.19The direction indicated for withdrawing or inserting the reservoir 1402 from the pipette station guard 1202 is different from the direction in which the locking hooks of the pipette station guard 1202 are configured to be retracted from engagement with the hook fasteners of the pipette station 1302. Such features can prevent accidental removal of components from the pipette docking set during electroporation, thereby improving user safety.

[0199] Example Port Gate

[0200] Fig. 20 A rear panel 2004 of a pulse generator 120 (eg, corresponding to pulse generator 2002) of an electroporation system is depicted. Fig. 20 The pulse generator 2002 is shown to include one or more connection ports (e.g., "USB port 1", "USB port 2", "LAN port", "WIFI port", etc.). Fig. 20 In the example of , each connection port has a corresponding port door 2006. As described above, the port door 2006 can improve user safety (eg, protect the user from high voltage discharge through low voltage components) and enable the pulse generator 2002 to meet applicable safety standards.

[0201] Fig.21 An isolated view of an example port gate 2006 of a pulse generator 2002 is provided. Fig.21 As shown, port door 2006 includes door frame 2102 and door 2104. Door 2104 is configured to translate vertically along door frame 2102 to facilitate selective opening of port door 2006 to selectively expose a connection port associated with port door 2006 (although other configurations, such as a hinged door, may be used). Fig.21 Port door 2006 is also shown to include a biasing element 2106 that biases door 2104 into a closed configuration to prevent access to a connection port associated with port door 2006. Although Fig.21 The biasing element 2106 is depicted as a torsion spring, but the biasing element 2106 may take any suitable form.

[0202] exist Fig.21 In the example of , the port door 2006 also includes a retaining wall 2108 for retaining the biasing element 2106 between the retaining wall 2108 and the rear panel 2004 of the pulse generator 2002 when the port door 2006 is connected to the rear panel 2004. Fig. 22 The port door 2006 is shown snap-fitted into connection with the rear panel 2004 (eg, from the inside of the rear panel 2004 ), with the biasing element 2106 disposed between the retaining wall 2108 and the rear panel 2004 .

[0203] Fig.21It is also shown that the port door 2006 can include at least one tool interface 2110 on its door 2104. The tool interface 2110 can be configured to receive a tool (e.g., a hex tool or any other type of tool) that a user can operate to selectively open the door 2104 to access a connection port associated with the port door 2006 (e.g., by using the tool to overcome the biasing force of the biasing element 2106 to move the door 2104 into an open configuration). One tool can be used with multiple port doors, and / or multiple tools can be associated with different port doors.

[0204] Fig.23 The tool 2302 is shown inserted into the tool interface 2110 of the door 2104 of the port door 2006. With the tool 2302 inserted into the tool interface 2110, the user can apply a force (e.g., a downward force in this example) to the tool 2302 to overcome the biasing force of the biasing element 2106 of the port door 2006. Fig.24A The port door 2104 is shown in an open configuration in response to a force applied to the door 2006 via the tool 2302 overcoming the biasing force of the biasing element 2106 of the port door 2006. Fig.24A As shown, when the port door 2006 is in the open configuration, the connection port 2402 associated with the port door 2006 is exposed.

[0205] Fig. 24B A connection component 2404 (e.g., a USB drive) is shown inserted into a connection port 2402 associated with the port door 2006. Inserting the connection component 2404 into the connection port 2402 while the port door is in the open configuration allows the connection component 2404 to remain reactive to the biasing force of the biasing element 2106 of the port door 2006, thereby allowing the port door 2006 to remain in the open configuration even after the tool 2302 is removed from the tool interface 2110. Fig.25A The connecting member 2404 is shown to maintain the open configuration of the port door 2006 without the tool 2302. When the port door 2006 is in the open configuration, the connecting member 2404 can be opened without the intervention of the tool 2302 (such as Fig.25A The disconnection of the connection member 2404 from the connection port 2402 eliminates the reaction force on the biasing element 2106 of the port door 2006, thereby allowing the biasing element 2106 to automatically force the port door 2006 back to the closed configuration (as indicated by the thick arrow in FIG. 24). Fig.25B ), without requiring additional user action to cause port door 2006 to close.

[0206] In some implementations, the tool 2302 for facilitating selective opening of the port door 2006 of the pulse generator 2002 can be conveniently mounted on the pulse generator 2002 (and / or on another component of the electroporation system). Fig. 20 Tool 2302 is shown mounted to tool holder 2022 of pulse generator 2002. Fig. 20 In the example of FIG. 2 , the tool 2302 is magnetically mounted to a tool holder 2022 disposed on a cable adapter 2020 of the pulse generator 2002. Additional details related to the cable adapter 2020 of the pulse generator 2002 will be provided below.

[0207] Example Cable Assembly

[0208] Fig.26A An isolated view of a cable adapter 2020 of a pulse generator 2002 is provided. The cable adapter 2020 is configured to hold various cables connected to the pulse generator 2002. For example (brief reference Fig. 20 ), the pulse generator 2002 may include a high-pressure connection port 2030 and a low-pressure connection port 2032. The high-pressure connection port 2030 may be configured to receive high-pressure cables, which may also be connected to the high-pressure port of the pipette station to facilitate electroporation. The low-pressure connection port 2032 may be configured to receive a low-pressure cable connected to the low-pressure port of the pipette station to facilitate various functions related to performing electroporation (e.g., process monitoring / execution, data acquisition, sensor operation / monitoring, etc.).

[0209] In this way, the high voltage cable and the low voltage cable can be selectively detached from the pulse generator 2002, thereby allowing the cables to be replaced in the event of a cable failure. In some cases, the high voltage connection port 2030 is a female connection port, which can improve user safety by reducing the possibility of the user accidentally contacting the electrodes of the high voltage connection port 2030.

[0210] The cable adapter 2020 can selectively retain at least a portion of a high voltage and / or low voltage cable configured to connect to the pulse generator 2002. Such functionality can be beneficial, for example, in allowing the pulse generator 2002 and components associated therewith to be stored or placed on a workstation (e.g., inside a biosafety cabinet) in a space efficient manner (e.g., particularly where the pulse generator 2002 is configured to connect via cables to multiple pipette stations and / or multiple external components).

[0211] exist Fig.26AIn the example of FIG. 2 , the cable adapter 2020 includes an external opening 2602 on (or at least partially defined by) a first surface 2604 (e.g., a rear surface) of the cable adapter 2020. The external opening 2602 exposes an interior of the cable adapter 2020 that can selectively retain at least a portion of a high voltage and / or low voltage cable associated with the pulse generator 2002. The external opening 2602 can allow a user to selectively place, rearrange, or remove the high voltage and / or low voltage cables from the interior of the cable adapter 2020.

[0212] Fig.26A The cable adapter 2020 also includes one or more cable troughs 2606 extending from the exterior opening 2602 and at least partially disposed on (or at least partially defined by) another surface 2608 of the cable adapter 2020. The cable troughs 2606 also expose the interior of the cable adapter 2020 and can provide a user with a convenient cable entry point into the interior cable adapter 2020 and / or a convenient exit point from the interior cable adapter for effective cable management.

[0213] As mentioned above, Fig.26B The cable adapter 2020 includes a tool holder 2022 that can hold a port door tool 2302. The cable adapter 2020 may include one or more finger-shaped recesses 2610 to allow a user to easily remove the tool 2302 from the tool holder 2022.

[0214] As further described above, the tool holder 2022 can be configured to magnetically hold the tool 2302 (which can be magnetizable or magnetic). Fig.26B Magnets 2612 are shown that can be disposed on the interior of the cable adapter 2020 (e.g., with a magnet cover 2614 placed thereon) to enable the tool 2302 to be secured by the magnets 2612 when placed within the tool holder 2022 of the cable adapter 2020. In view of this disclosure, it should be understood that other types of attachment methods for securing the tool 2302 within the tool holder 2022 are within the scope of this disclosure.

[0215] like Fig.26B As shown, the cable adapter 2020 may include a mounting feature 2620 to facilitate selective mounting of the cable adapter 2020 to the pulse generator 2002. Fig.26BIn the example of , mounting feature 2620 includes a disk 2622 positioned on a mounting arm 2624 extending from a surface of cable adapter 2020. Other configurations and / or types of mounting features may be used. Disk 2622 may include a larger radius or lateral width / thickness than mounting arm 2624 to facilitate engagement with a corresponding mounting hole of pulse generator 2002. Fig.26B It is also shown that the cable adapter 2020 may include additional features associated with mounting the cable adapter 2020 to the pulse generator 2002, such as alignment features 2626.

[0216] Fig.27A A cross-sectional view of a cable adapter 2020 and a pulse generator 2002 is shown. As shown, the pulse generator 2002 includes a mounting hole 2702 for receiving the mounting feature 2620 of the cable adapter 2020. In the example of FIG. 27 , the mounting hole 2702 is defined by a pair of conjoined holes, wherein a first (top) hole has a larger radius than a second (bottom) hole disposed below the first hole. The radius of the first hole is larger than the radius of the disk 2622 of the mounting feature 2620 of the cable adapter 2020. The second hole has a smaller radius than the first hole and the disk 2622 of the mounting feature 2620 of the cable adapter 2020. The radius of the second hole is larger than the radius of the mounting arm 2624 of the mounting feature 2620 of the cable adapter 2020.

[0217] Thus, the disk 2622 can be inserted into the mounting hole 2702 through the first hole, as shown in FIG. Fig.27A As shown by the thick arrows and Fig.27B After being inserted therethrough, the cable adapter 2020 can be allowed to fall until the mounting arm 2624 of the mounting feature 2620 rests on the second hole of the mounting hole 2702, as shown. Fig.27C The disk 2622 can then prevent the cable adapter 2020 from being removed from the pulse generator 2002 until the cable adapter is lifted to position the disk toward the first hole of the mounting hole 2702.

[0218] like Fig.27AAs shown, the alignment features 2626 of the cable adapter 2020 can be mated with corresponding alignment features 2704 (implemented as elongated channels or slots in the illustrated example) of the pulse generator 2002. When the disk 2622 is inserted through the mounting hole 2702 and the alignment features 2626 are mated with the corresponding alignment features 2704, such mating can force the cable adapter 2020 to be in a specific alignment (e.g., vertical alignment in the illustrated example) with the pulse generator 2002. Thus, when the mounting features 2620 of the cable adapter 2020 are mounted to the mounting hole 2702 of the pulse generator, the alignment features 2704 can define the final position of the cable adapter 2020 relative to the pulse generator (thereby mitigating accidental displacement or rotation of the cable adapter 2020 when mounted to the pulse generator 2002).

[0219] Fig.28A An example cable assembly 2802 that can be used in an electroporation system is shown. The cable assembly 2802 may include a Fig.28A and includes at the opposite end a component for connecting to a pipette station (in Fig.28A The parts marked as "STATION" in the figure. Fig.28B An example aspect of components for connecting to a pulse generator is shown, which may include a high voltage connector 2804 for a high voltage cable 2806 and a low voltage connector 2808 for a low voltage cable 2810. Similarly, Fig.28C Example aspects of components for connecting to a pipette station are shown, which may include a corresponding high voltage connector 2812 for the high voltage cable 2806 and a corresponding low voltage connector 2814 for the low voltage cable 2810 .

[0220] Fig.28A It is also shown that the cables of the cable assembly 2802 (e.g., the high voltage cable 2806 and the low voltage cable 2810) can be tied or connected to each other via the tie assembly 2820. Fig.28A In the example of FIG. 2 , the binding assembly 2820 includes a braided sleeve 2822 that surrounds at least a section of the high voltage cable 2806 and the low voltage cable 2810 . Fig.28A The braided sleeve 2822 is connected to a clamp 2830, which also surrounds the high voltage cable 2806 and the corresponding section of the low voltage cable. The clamp 2830 can be formed of any suitable material, such as a polymer material.

[0221] In some implementations, each of the clamps 2830 is formed from multiple parts that are attached to each other around the high voltage cable 2806 and the low voltage cable 2810 . Fig.29AA first part 2902 and a second part 2904 are shown forming one of the fixtures 2830. When attached to each other, as shown in FIG. Fig.29B As shown, the first part 2902 and the second part 2904 form a plurality of conjoined holes 2906 that can surround the high voltage cable 2806 and the low voltage cable 2810. The first part 2902 and the second part 2904 can be combined in a variety of ways, such as by ultrasonic welding.

[0222] Example Aspects of Electrical Pulse Application

[0223] Fig.30 A schematic diagram of electric pulses applied using an electroporation system 3000 (e.g., corresponding to electroporation system 100) is shown, which includes a pipette station 3002 (e.g., corresponding to pipette station 1302), a pipette station guard 3004 mounted to the pipette station 3002 (e.g., corresponding to pipette station guard 1202), a buffer reservoir 3006 mounted to the pipette station guard 3004 (e.g., corresponding to buffer tube 1402), and a pipette 3008 (corresponding to pipette 820 having a pipette tip 802 attached thereto) inserted into the buffer tube 3006.

[0224] Fig.30 The pipette station 3002 includes electrodes 3010 and 3012 that can be electrically connected to the pulse generator via high voltage cables and / or ports, as discussed above. The buffer tube 3006 also includes a buffer reservoir electrode 3014 (e.g., corresponding to electrode 1418) that is exposed to the interior of the buffer tube 3006 (e.g., the interior of the buffer reservoir 3016 of the buffer tube 3006) and is configured to contact the electrode 3012 of the pipette station 3002 when the buffer tube is installed to the pipette station guard 3004 of the pipette station 3002. Thus, the buffer reservoir electrode 3014 allows electrical pulses from the pulse generator to reach the buffer solution 3018 within the buffer reservoir 3016 of the buffer tube 3006 (e.g., by completing a circuit in which the electrical pulse passes through the pipette 3008 to the sample within the pipette tip 3024 and returns through the buffer solution 3018 and the buffer reservoir electrode 3014).

[0225] Buffer tube 3006 also includes an electrode opening (with Fig.143002), through which a pipette electrode 3020 of the pipette 3008 can extend when the pipette 3008 is mounted to the buffer tube 3006 (which is mounted to the pipette station guard 3004, which is mounted to the pipette station 3002) to allow the pipette electrode 3020 to contact the electrode 3010 of the pipette station 3002. The pipette electrode 3020 is in electrical communication with a plunger 3022 of a pipette tip 3024 associated with the pipette 3008, and the plunger 3022 is configured to contact any sample disposed within the pipette tip 3024.

[0226] Thus, the pulse generator can provide one or more high voltage pulses to the sample within the pipette tip 3024 to electroporate the sample when the following conditions occur: (i) the pipette tip 3024 is connected to the pipette 3008 and is disposed within the buffer solution 3018 to expose the sample within the pipette tip 3024 to the buffer solution 3018 at the tip opening of the pipette tip 3024, (ii) the buffer reservoir electrode 3014 is in contact with the buffer solution 3018 and the electrode 3012 of the pipette station 3002, (iii) the electrode 3010 of the pipette station 3002 is in contact with the pipette electrode 3020, and (iv) the pipette electrode 3020 is electrically connected to the plunger 3022 of the pipette tip, which is in contact with the sample in the pipette tip.

[0227] In some implementations, the electroporation system includes one or more sensors for determining whether the station guard 3004, the buffer tube 3006, and / or the pipette are properly positioned relative to each other to prevent high voltage discharge unless these components are properly interconnected. Such sensors can take any suitable form, and the sensor data obtained thereby can be transmitted via a low voltage cable.

[0228] As described above, conventional electroporation systems utilize an open-loop pulse generator that uses a charging circuit to charge a large capacitor to a target voltage and then uses a high-speed, high-voltage electronic switch to connect the voltage at the capacitor to the pipette station. In some cases (e.g., when the load resistance is small and / or for long pulse durations), the voltage reaching the sample begins to drop during the pulse duration (e.g., because the charging electronics fail to provide sufficient power to keep the load at the target voltage). As a result, a voltage drop may occur, such as Fig.31 As shown (for a target voltage of 2.5 kV).

[0229] The pulse generators of the present disclosure (e.g., pulse generator 120) can utilize feedback loop control in which a capacitor is charged to a voltage above a target voltage and the voltage is adjusted as needed to produce a stable voltage supply at a desired voltage (and / or pulse width and / or modulation setting). Fig.32An example flat pulse waveform that may be applied using a pulse generator according to the present disclosure is shown.

[0230] In some embodiments, the pulse generator includes one or more voltage sources configured to charge one or more high-voltage capacitors. The high-voltage capacitor is configured to be used as a power source for an amplifier circuit. The amplifier circuit is configured to supply voltage to a sample associated with the pipette in a manner that takes into account load variations. For example, the load can vary for different reaction conditions (e.g., buffer solution type, pipette tip size, cell concentration, etc.) and / or throughout the electroporation process (e.g., based on temperature changes).

[0231] The amplifier circuit may include a common source amplifier configured to output a high voltage pulse. The common source amplifier may receive a signal from an amplitude setting loop. The signal of the amplitude setting loop is based on an input from a digital-to-analog converter and an input from a voltage sensing loop. The input from the digital-to-analog converter may correspond to a waveform selected by a user (e.g., a sine wave, a triangle wave, a square wave, etc.). The input from the voltage sensing loop is determined using a high voltage pulse, a voltage divider, and a differential amplifier. The common source amplifier may amplify the signal from the amplitude setting loop by about 1,000 times to about 2,000 times (e.g., 1,250 times).

[0232] Fig.33 An example architecture that can be implemented in a pulse generator of an electroporation system according to a specific implementation of the present disclosure is shown. As shown, the circuit design for pulse generation uses a voltage divider and a differential amplifier (e.g., a "voltage sensing loop") to convert the HV voltage to V_SENSE (e.g., HV_Voltage / 1250). The pulse waveform output from the DAC (digital-to-analog converter) to V_DAC is compared and amplified with V_SENSE to facilitate low-side switching of the high-voltage MOSFET (e.g., an "amplitude setting loop"). Based on the amplitude of the high-voltage output at HV_Voltage, the V_DAC amplitude is set to HV_Voltage / 1250, and the waveform at the V_DAC sets the waveform at the high-voltage output at the port HV+.

[0233] The pulse generator architecture as currently disclosed may enable waveform customization, allowing for implementation of sine, square, triangle, sawtooth, and / or other waveforms.

[0234] Fig.34 A simulation-based integrated circuit (SPICE model or simulation program model with integrated circuit emphasis) is shown with optimized component values ​​(eg, resistors, capacitors). The circuit shown is designed to produce a flat pulse. Figure 35 to Figure 37 Example waveform simulations of different simulation waveforms implementing the disclosed architecture are shown. Fig.35An example sine wave is shown with an input (eg, VG1 ) of 1 kHz frequency, 1 V amplitude, and 1 V offset, and a corresponding output (eg, VM1 ) of 1 kHz frequency, 1250 V amplitude, and 1250 V offset. Fig.36 An example triangle wave is shown with an input (eg, VG1 ) of 1 kHz frequency, 1 V amplitude, and 1 V offset, and a corresponding output (eg, VM1 ) of 1 kHz frequency, 1250 V amplitude, and 1250 V offset. Fig.37 An example sine wave is shown with an input (eg, VG1 ) of 1 kHz frequency, 1 V amplitude, and 1 V offset, and a corresponding output (eg, VM1 ) of 1 kHz frequency, 1250 V amplitude, and 1250 V offset.

[0235] Example Arc Detection Module

[0236] As described above, during high voltage electroporation (500V to 2500V), if any bubbles are introduced into the tip holding the cell sample, an arc may occur. The arc may lead to electroporation failure and / or poor transfection results. Traditionally, arc detection relies on human observation, which may lead to detection errors and / or failures. Therefore, at least some disclosed embodiments implement an arc detection module that is configured to detect arcs without relying on human observation.

[0237] Detection of a sudden drop in the electroporation pulse current can indicate arcing. Fig.38 Example voltage and current waveforms of an electroporation pulse in which no arcing occurs are shown. Fig.39 Example voltage and current waveforms of an electroporation pulse are shown, illustrating the sudden drop in current associated with arcing.

[0238] Because different reaction conditions may exist, it is beneficial for the arc detection module to be able to detect a current drop for different types of tips, buffers, load resistors (which may change based on temperature throughout the electroporation process), etc. Therefore, an arc detection module configured to detect an arc during electroporation may include (i) a first stage amplifier configured to provide an amplified current signal based on a current signal associated with a voltage applied to the sample (the amplification of the current signal by the first stage amplifier may be based on the output of a low voltage detection circuit used to determine the resistance associated with the sample), (ii) a band pass filter configured to filter a falling edge signal from the amplified current signal (where the falling edge signal indicates a drop in current through the sample, which indicates an arc), and (iii) a comparator configured to compare the falling edge signal filtered by the band pass filter with one or more reference standards to determine whether an arc has occurred in the sample.

[0239] Fig.40An example first stage amplifier for an arc detection module implemented in an electroporation system is shown. As shown, the pulsed electroporation current ("I_SENSE") is sensed by a resistor (e.g., a 3 ohm resistor) and converted to a voltage. A microcontroller unit (MCU) can provide an H / L signal ("Arc_Amp_gain") to adjust the gain of the first stage amplifier based on the type of pipette tip (e.g., when a 10 μL pipette tip is used, the gain can be 10 times relative to the gain for a 100 μL pipette tip).

[0240] Fig.41 Shows Fig.40 An example of an arc detection module with a bandpass filter and comparator. Fig.40 The first stage amplifier can be connected to Fig.41 A bandpass filter such as Fig.40 The thick arrows marked "to bandpass filter" and Fig.41 This is shown by the thick arrow labeled “from first amplifier stage” in the figure.

[0241] The bandpass filter can remove the normal current waveform and output the sudden current drop signal (if present) to the comparator. The comparator circuit can then determine whether the current fault signal corresponds to an arc event (e.g., based on a comparison with a reference signal / reference data). The output of the arc detection circuit can include a logic signal ("ARCING") that is connected to a microprocessor unit (MCU) as an input to indicate whether an arc is detected.

[0242] Fig.42 A simulation-based integrated circuit (SPICE model) is shown that optimizes component values ​​(e.g., resistors, capacitors, etc.). The circuit shown is designed to detect arcs with high sensitivity while avoiding false detections. The values ​​of the components can be selected based on part availability. The input of the current curve is simplified to a sudden drop within 1 millisecond. Different starting currents (e.g., the current starts to drop from this starting current) are set to simulate different buffer types and different pipette tip types to be used. Fig.43 The simulation results show that the voltage (I_SENSE) drops from 2.5V to 1.78V when an arc occurs and the arc signal output increases accordingly. As described above, the arc signal can be sent to the MCU.

[0243] Figures 44 to 46 Test results from tests performed to verify the operation of the arc detection circuit described above are shown. The tests were performed at different voltage settings from 2,500 V to 500 V with different pulse widths and various numbers of pulses. When a sudden drop in current occurs due to arcing during electroporation, the test results indicate a strong ARCING signal.

[0244] Additional computer system details

[0245] The disclosed embodiments may include or utilize a special or general-purpose computer including computer hardware, as discussed in more detail below. The disclosed embodiments also include physical and other computer-readable media for carrying or storing computer-executable instructions and / or data structures. Such computer-readable media may be any available media that can be accessed by a general or special-purpose computer system. A computer-readable medium that stores computer-executable instructions in the form of data is one or more "physical computer storage media" or "hardware storage devices." A computer-readable medium that only carries computer-executable instructions but does not store computer-executable instructions is a "transmission medium." Therefore, by way of example and not limitation, the current embodiments may include at least two distinct types of computer-readable media: computer storage media and transmission media.

[0246] Computer storage media (also called "hardware storage devices") are computer-readable hardware storage devices based on RAM, flash memory, phase change memory ("PCM"), or other types of memory, such as RAM, ROM, EEPROM, CD-ROM, solid-state drive ("SSD"), or other optical disk storage, magnetic disk storage or other magnetic storage devices, or any other medium that can be used to store the desired program code tools in the form of computer-executable instructions, data or data structures in hardware and that can be accessed by a general or special purpose computer.

[0247] A "network" is defined as one or more data links that enable the transmission of electronic data between computer systems and / or modules and / or other electronic devices. When information is transferred or provided to a computer over a network or other communications connection (hardwired, wireless, or a combination of hardwired or wireless), the computer properly views the connection as a transmission medium. Transmission media may include networks and / or data links that may be used to carry program code in the form of computer-executable instructions or data structures and that may be accessed by general-purpose or special-purpose computers. Combinations of the foregoing are also included within the scope of computer-readable media.

[0248] Furthermore, program code tools in the form of computer executable instructions or data structures may be automatically transferred from a transmission computer readable medium to a physical computer readable storage medium (and from a physical computer readable storage medium to a transmission computer readable medium) upon reaching various computer system components. For example, computer executable instructions or data structures received over a network or data link may be cached in RAM within a network interface module (e.g., a "NIC") and then ultimately transferred to computer system RAM and / or a less volatile computer readable physical storage medium at the computer system. Thus, a computer readable physical storage medium may be included in a computer system component that also (or even primarily) utilizes a transmission medium.

[0249] Computer executable instructions include, for example, instructions and data that cause a general purpose computer, a special purpose computer, or a special purpose processing device to perform a certain function or group of functions. Computer executable instructions may be, for example, binary, intermediate format instructions such as assembly language, or even source code.

[0250] The disclosed embodiments may include or utilize cloud computing. Cloud models may include various features (e.g., on-demand self-service, broad network access, resource pooling, rapid elasticity, measured services, etc.), service models (e.g., software as a service ("SaaS"), platform as a service ("PaaS"), infrastructure as a service ("IaaS")), and deployment models (e.g., private cloud, community cloud, public cloud, hybrid cloud, etc.).

[0251] Those skilled in the art will appreciate that the present invention can be practiced in a network computing environment with many types of computer system configurations, including personal computers, desktop computers, laptop computers, message processors, handheld devices, multiprocessor systems, microprocessor-based or programmable consumer electronics, network PCs, minicomputers, mainframe computers, mobile phones, PDAs, pagers, routers, switches, wearable devices, etc. The present invention can also be implemented in a distributed system environment, where multiple computer systems (e.g., local and remote systems) linked by a network (by a hardwired data link, a wireless data link, or a combination of hardwired and wireless data links) perform tasks. In a distributed system environment, program modules can be located in local and / or remote memory storage devices.

[0252] Alternatively or additionally, the functions described herein may be performed at least in part by one or more hardware logic components. For example, but not limited to, illustrative types of hardware logic components that may be used include field programmable gate arrays (FPGAs), application specific integrated circuits (ASICs), application specific standard products (ASSPs), systems on chips (SOCs), complex programmable logic devices (CPLDs), central processing units (CPUs), graphics processing units (GPUs), and / or other components.

[0253] As used herein, the terms "executable module," "executable component," "component," "module," or "engine" may refer to a hardware processing unit or a software object, routine, or method that can be executed on one or more computer systems. The various components, modules, engines, and services described herein may be implemented as objects or processors (e.g., as separate threads) that execute on one or more computer systems.

[0254] In some embodiments, the system of the present disclosure may include or may be configured to execute any combination of software and / or hardware components that are operable to facilitate processing using machine learning models or other artificial intelligence-based structures / architectures. For example, one or more processors may include and / or utilize hardware components and / or computer executable instructions to execute functional blocks and / or processing layers configured in the following form, as non-limiting examples, single-layer neural networks, feedforward neural networks, radial basis function networks, deep feedforward networks, recurrent neural networks, long short-term memory (LSTM) networks, gated recurrent units, autoencoder neural networks, variational autoencoders, denoising autoencoders, sparse autoencoders, Markov chains, Hopfield neural networks, Boltzmann machine networks, Boltzmann machine networks, deep belief networks, deep convolutional networks (or convolutional neural networks), deconvolutional neural networks, deep convolutional inverse graph networks, generative inverse networks, liquid machines, extreme learning machines, echo state networks, deep residual networks, Kohonen networks, support vector machines, neural Turing machines, and / or others.

[0255] It will also be understood that the systems, processes and / or products according to certain embodiments of the present disclosure may include, incorporate or otherwise include the characteristic features (e.g., components, members, elements, parts and / or parts) described in other embodiments disclosed and / or described herein. Therefore, the various features of certain embodiments may be compatible, combined, incorporated and / or incorporated into other embodiments of the present disclosure with other embodiments of the present disclosure. Therefore, certain features are disclosed with reference to the specific embodiments of the present disclosure, which should not be considered as limiting the application of the features to or inclusion in the specific embodiments. On the contrary, it should be understood that other embodiments may also include the features without necessarily departing from the scope of the present disclosure.

[0256] In addition, unless a feature is described as being required to be combined with another feature, any feature herein may be combined with any other feature of the same or different embodiments disclosed herein. In addition, various well-known aspects of exemplary systems, processes, products, etc. are not described in particular detail herein to avoid obscuring various aspects of the example embodiments. However, such aspects are also contemplated herein.

[0257] Although the present disclosure provides certain illustrative aspects and describes the general principles of the described technology, it will be understood by those skilled in the relevant art that modifications to the arrangements and details of the present disclosure may be introduced without departing from these aspects and principles. Therefore, the applicant claims all modifications within the spirit and scope of the appended claims.

Claims

1. A pipette, the pipette include: a proximal segment having a handle; a distal segment configured to be reversibly attached to a pipette tip; a first actuator disposed in the proximal section, the first actuator being operable, when actuated, to control: i) a pipetting function of the pipette; and ii) gripping and releasing of a plunger disposed within an inner cavity of the pipette tip; and A second actuator is disposed in the proximal section and is operable, when actuated, to detach the pipette tip from the distal section of the pipette.

2. The pipette of claim 1, further comprising a pipette electrode disposed in the distal section and electrically coupled to the plunger when grasped.

3. A pipette according to claim 1, wherein the pipetting function includes aspirating fluid into a pipette tip attached to the distal segment or dispensing fluid from the pipette tip.

4. A pipette according to claim 3, wherein the first actuator has a first undepressed position and a second partially depressed position, and wherein transitioning the first actuator from the first undepressed position to the second partially depressed position causes dispensing from the pipette tip, and transitioning the first actuator from the second partially depressed position to the first undepressed position causes aspiration into the pipette tip.

5. The pipette of claim 4, wherein the first actuator has a third fully depressed position.

6. A pipette according to claim 5, wherein transitioning the first actuator from the second partially depressed position to the third fully depressed position causes release of the plunger, and transitioning the first actuator from the third fully depressed position to the second partially depressed position causes gripping of the plunger.

7. The pipette according to claim 6 further comprises a clamp mechanism disposed in the distal segment, the clamp mechanism being operable to transition between a closed configuration and an open configuration when the first actuator transitions between the second partially depressed position and the third fully depressed position.

8. The pipette of claim 7, wherein the gripper mechanism include: a holder jaw including a jaw opening for receiving an engagement segment of the plunger; and a clamping sleeve positioned around the clamp jaws, the clamping sleeve being configured to apply an inward force to the clamp jaws so that the clamp jaws apply a compressive force to the engagement segment of the plunger to retain the engagement segment of the plunger within the jaw opening when the first actuator is in the first depressed position or the second partially depressed position.

9. A pipette according to claim 8, wherein the clamp jaws and the clamping sleeve are constructed to translate within the pipette while retaining the engagement section of the plunger within the jaw opening so that the inner cavity section of the plunger translates within the inner cavity of the pipette tip to facilitate the pipetting function.

10. The pipette of claim 9, wherein when the first actuator transitions from the second partially undepressed position to the third fully depressed position, the gripper jaw moves distally relative to the gripper sleeve, thereby releasing the plunger from the gripper jaw.

11. The pipette of claim 10, wherein the second actuator has a first, undepressed position and a second, depressed position.

12. A pipette according to any of the preceding claims, further comprising a tip interface disposed circumferentially around the jaws of the holder, the tip interface comprising a retaining platform configured to engage a tab of an attachment interface of the pipette tip to secure a tip sleeve defining the inner cavity of the pipette tip to the distal segment of the pipette.

13. The pipette according to any of the preceding claims, further comprising a tip ejection sleeve operably connected to the second actuator and disposed adjacent to the tip interface, the tip ejection sleeve being operable to move distally relative to the tip interface when the second actuator is actuated by transitioning the second actuator from the first undepressed position to the second depressed position, and to displace the attachment interface of the pipette tip from the retaining platform to separate the tip sleeve from the pipette.

14. The pipette of any one of the preceding claims, wherein the second actuator is configured to traverse a blank travel distance when depressed prior to detaching one or more tabs from the retaining platform.

15. The pipette of claim 14, wherein the gripper mechanism is configured to retain the engagement segment of the plunger within a gripper opening throughout separation of the attachment interface from the retention platform.

16. The pipette according to claim 15, in, When the attachment interface and tip sleeve of the pipette tip are separated from the distal section of the pipette, transitioning the first actuator from the second partially depressed position to the third fully depressed position causes the release of the engagement section of the plunger held by the clamp jaws, thereby releasing the pipette tip from the distal section of the pipette.

17. The pipette of any of the preceding claims, wherein the pipette electrode is electrically coupled to the gripper jaws of the gripper mechanism.

18. A pipette according to claim 17, wherein the clamp jaws are made of a conductive material and are capable of being operated to allow an electrical pulse applied to the pipette electrode to pass through the pipette electrode, through the clamp jaws, through the plunger retained in the jaw opening of the clamp jaws, through the cell-containing sample contained in the inner cavity of the pipette tip, and through a second electrode disposed adjacent the distal end of the pipette tip, thereby electroporating the cells contained in the sample.

19. The pipette of any of the preceding claims, wherein the pipette comprises 2, 3, 4, 5, 6, 7, 8 or more gripper mechanisms operably connected to the first actuator, 2, 3, 4, 5, 6, 7, 8 or more tip ejection sleeves operably connected to the second actuator, and 2, 3, 4, 5, 6, 7, 8 or more tip interfaces.

20. The pipette of any of the preceding claims, further comprising a pipette tip attached to the distal section of the pipette via the tip sleeve and / or the plunger.

21. An electroporation system, the electroporation system include: The pipette according to any one of claims 1 to 20; Pipette tips; Pipette docking assembly; and Pulse generator.

22. The electroporation system of claim 21, wherein the pipette docking assembly comprises a pipette station, a pipette station guard, and a reservoir.

23. The electroporation system of claim 22, wherein the reservoir comprises a buffer.

24. The electroporation system of any one of claims 21 to 23, wherein the pipette tip has a sample volume capacity of between 10 μL and 100 μL.

25. The electroporation system of claim 24, wherein the pipette tip has a sample volume capacity of 10 μL or 100 μL.

26. An electroporation system according to any one of claims 21 to 25, wherein the pipette tip comprises a plunger at least partially disposed within an inner cavity defined by a tip sleeve, the plunger being configured to translate along the inner cavity to facilitate aspiration and / or dispensing.

27. The electroporation system of claim 26, wherein the plunger comprises gold, diamond-like carbon, and / or conductive plastic.

28. An electroporation system according to claim 26 or claim 27, wherein the plunger comprises an engagement section and an inner cavity section, the inner cavity section comprising a sealing component for creating a seal between the plunger and the inner cavity wall of the pipette tip sleeve.

29. The electroporation system of claim 28, wherein the lumen segment comprises a front pin and a shaft segment, wherein the front pin is configured to connect to the shaft segment and secure the sealing component to the shaft segment.

30. The electroporation system of claim 29, wherein the sealing component comprises a polymer sleeve, and wherein the front pin is configured to be inserted through the polymer sleeve and engage the retaining hole of the shaft segment to secure the sealing component to the shaft segment.

31. The electroporation system of claim 30, wherein insertion of the front pin through the polymer sleeve defines a space between the polymer sleeve and the front pin, and wherein the space contributes to flexibility of the polymer sleeve for creating the seal between the plunger and the lumen.

32. The electroporation system of any one of claims 28 to 31, wherein the sealing member comprises polytetrafluoroethylene (PTFE).

33. The electroporation system of claim 28, wherein the sealing component comprises a coated O-ring, and wherein the lumen segment comprises a circumferential recess configured to receive the coated O-ring.

34. An electroporation system according to any one of claims 28 to 33, wherein the pipette tip further comprises an attachment interface adjacent to the inner cavity, the attachment interface comprising one or more tabs configured to engage with a retaining platform of a distal segment of the pipette.

35. The electroporation system of any one of claims 22 to 34, wherein the pipette station guard comprises a reservoir opening for receiving the reservoir.

36. An electroporation system according to claim 35, wherein the pipette station includes one or more locking hooks, which are configured to engage with one or more corresponding hook fasteners of the pipette station, and wherein the one or more locking hooks are configured to retract from the engagement with the one or more corresponding hook fasteners of the pipette station in one or more directions that are at least partially different from the following directions: i) a reservoir withdrawal direction for withdrawing the reservoir from the reservoir opening of the pipette station guard; and / or ii) a pipette withdrawal direction for withdrawing the pipette from the reservoir when the reservoir is positioned in the reservoir opening of the pipette station guard.

37. An electroporation system according to claim 36, wherein the one or more locking hooks include one or more pivoting hooks, and the one or more pivoting hooks are configured to rotate into engagement with at least some of the one or more corresponding hook fasteners of the pipette station.

38. An electroporation system according to claim 36 or claim 37, wherein the one or more locking hooks include one or more flexible hooks, and the one or more flexible hooks are configured to advance to engage with at least some of the one or more corresponding hook fasteners of the pipette station.

39. An electroporation system according to claim 38, wherein the pipette station guard further comprises one or more finger guides, the one or more finger guides indicating one or more pressable surfaces of the pipette station guard, the one or more pressable surfaces being pressable to facilitate the disengagement of the one or more flexible hooks from at least some of the one or more corresponding hook fasteners of the pipette station.

40. An electroporation system according to claim 39, wherein the one or more locking hooks include a pair of pivoting hooks and a pair of flexible hooks, the pair of pivoting hooks are arranged on the top portion of the rear surface of the pipette station guard, the pair of flexible hooks are positioned on the bottom portion of the rear surface of the pipette station guard, and the one or more finger guides are arranged on the bottom portion of the side surface of the pipette station guard.

41. An electroporation system according to any one of claims 22 to 40, wherein the reservoir comprises one or more latching members, and the one or more latching members are configured to engage with one or more corresponding latching fasteners of the pipette station guard to secure the reservoir to the pipette station guard when the reservoir is inserted into the reservoir opening / the reservoir opening of the pipette station guard.

42. The electroporation system of claim 41, wherein the reservoir comprises one or more hooks for holding the pipette when the pipette is inserted into the reservoir.

43. An electroporation system according to claim 41 or claim 42, wherein one or more force application directions for disengaging the one or more latching members from the one or more corresponding latch fasteners are different from the pipette withdrawal direction / the pipette withdrawal direction for withdrawing the pipette from the reservoir when the reservoir is positioned in the reservoir opening of the pipette station guard.

44. An electroporation system according to claim 42 or claim 43, wherein the one or more latching members are disposed on an outer surface of the reservoir.

45. An electroporation system according to any one of claims 41 to 44, wherein the one or more latching members each include a corresponding finger prompt for guiding the positioning of a user's finger to apply force to the one or more latching members to facilitate the disengagement of the one or more latching members from the one or more corresponding latch fasteners.

46. ​​An electroporation system according to any one of claims 41 to 45, wherein the one or more latching members extend from a flange of the reservoir, the flange extending around a pipette opening of the reservoir to receive a distal segment of the pipette.

47. An electroporation system according to any one of claims 41 to 46, wherein the pipette station includes an interlocking feature, which is constructed to engage with a portion of the reservoir when the one or more latching members of the reservoir are engaged with the one or more corresponding latching fasteners of the pipette station guard and when the pipette station guard is connected to the pipette station, thereby preventing the pipette station guard from being removed from the pipette station when the interlocking feature is engaged with the portion of the reservoir.

48. An electroporation system according to any one of claims 21 to 47, wherein the pulse generator comprises one or more connection ports, each specific connection port among the one or more connection ports comprises a corresponding port door, and the corresponding port door comprises a corresponding biasing element for biasing the corresponding port door into a closed configuration to prevent access to the specific connection port.

49. An electroporation system according to claim 48, wherein the corresponding port door includes a corresponding tool interface constructed to receive a port door tool, and the port door tool is constructed to interact with the corresponding tool interface to react against the corresponding biasing element of the corresponding port door to bring the corresponding port door into an open configuration to provide access to the specific connection port.

50. The electroporation system of claim 49, wherein insertion of a port connection component into the particular connection port maintains the reaction against the corresponding biasing element to maintain the corresponding port door in the open configuration.

51. The electroporation system of claim 50, wherein disconnection of the port connection component from the particular connection port eliminates the reaction to the corresponding biasing element to allow the corresponding port door to return to the closed configuration.

52. An electroporation system according to any one of claims 49 to 51, wherein the pulse generator comprises a tool holder for holding the port door tool.

53. The electroporation system of claim 52, wherein the tool holder is disposed on a cable adapter of the pulse generator.

54. The electroporation system of claim 53, wherein the tool holder of the cable adapter comprises one or more magnets for facilitating retention of the port door tool on the cable adapter.

55. An electroporation system according to any one of claims 52 to 54, wherein the pulse generator comprises one or more high-pressure connection ports and one or more low-pressure connection ports, the one or more high-pressure connection ports being configured to receive one or more high-pressure cables to connect the one or more high-pressure connection ports to one or more corresponding high-pressure connection ports of the pipette station, and the one or more low-pressure connection ports being configured to receive one or more low-pressure cables to connect the one or more low-pressure connection ports to one or more corresponding low-pressure connection ports of the pipette station.

56. An electroporation system according to claim 55, wherein the pulse generator includes a cable adapter / the cable adapter, which is configured to selectively retain at least a portion of the one or more high-voltage cables and / or at least a portion of the one or more low-voltage cables.

57. An electroporation system according to claim 56, wherein the cable adapter includes an external opening on a first surface of the cable adapter, the external opening exposing the interior of the cable adapter for selectively retaining at least the portion of the one or more high-voltage cables and / or at least the portion of the one or more low-voltage cables.

58. The electroporation system of claim 57, wherein the cable adapter further comprises a cable trough extending from the external opening to a second surface of the cable adapter, the cable trough exposing the interior of the cable adapter, the one or more high voltage cables and / or the one or more low voltage cables being configured to extend through the cable trough into the interior of the cable adapter.

59. The electroporation system of any one of claims 56 to 58, wherein the cable adapter further comprises a mounting feature configured to interface with a mounting hole of the pulse generator.

60. The electroporation system of claim 59, wherein the mounting feature comprises a disc positioned on a mounting arm, and wherein the mounting aperture is defined by a pair of conjoined apertures comprising a first aperture having a first radius and a second aperture having a second radius.

61. An electroporation system according to claim 60, wherein the first hole is arranged above the second hole, and wherein the first radius is greater than the disc radius of the disc, and wherein the second radius is less than the disc radius but greater than the radius of the mounting arm, thereby allowing the disc to enter through the first hole to allow the mounting arm to reside on the second hole.

62. An electroporation system according to claim 61, wherein the cable adapter further includes an alignment feature, and wherein the pulse generator includes a corresponding alignment feature, and the alignment feature and the corresponding alignment feature are constructed to engage with each other when the mounting feature of the cable adapter is mounted to the mounting hole of the pulse generator to define the final positioning of the cable adapter relative to the pulse generator.

63. The electroporation system of any one of claims 55 to 62, wherein the one or more high voltage cables and the one or more low voltage cables are tied to each other via a tying assembly.

64. The electroporation system of claim 63, wherein the ligation assembly comprises a braided sleeve connected to one or more clamps.

65. An electroporation system according to claim 64, wherein each of the one or more clamps comprises a first part, which is attached to a second part around the one or more high-voltage cables and the one or more low-voltage cables, wherein the first part and the second part form a plurality of conjoined holes when attached to each other, and the plurality of conjoined holes are configured to receive the one or more high-voltage cables and the one or more low-voltage cables.

66. An electroporation system according to any one of claims 21 to 65, in: The reservoir includes a reservoir and a reservoir electrode exposed to the interior of the reservoir and the exterior of the reservoir / the exterior, the pipette station comprising a first electrode configured to contact the reservoir electrode when the reservoir is inserted into / into the reservoir opening of the pipette station guard and when the pipette station guard is connected to the pipette station, The side wall of the reservoir defines an electrode opening for allowing a pipette electrode of the pipette to contact a second electrode of the pipette station when the distal segment of the pipette / the distal segment is inserted into the pipette opening / the pipette opening of the reservoir, and the first electrode and the second electrode are configured to receive a high voltage pulse from the pulse generator to facilitate electroporation of a sample within the lumen / the lumen of the pipette tip when: The pipette tip is attached to the pipette, positioned within the reservoir, and in contact with a buffer within the reservoir, The first electrode is in contact with the reservoir electrode, the plunger of the pipette tip / the plunger is in electrical communication with the sample and the pipette electrode, and The pipette electrode is in contact with the second electrode.

67. An electroporation system according to any one of claims 21 to 66, wherein the pulse generator comprises one or more voltage sources configured to charge one or more high voltage capacitors, the one or more high voltage capacitors being configured to be used as a power source for an amplifier circuit, the amplifier circuit being configured to supply voltage to the sample of the pipette / the sample in a manner that takes into account load variations.

68. The electroporation system of claim 67, wherein the amplifier circuit comprises a common source amplifier configured to output high voltage pulses.

69. An electroporation system according to claim 68, wherein the common source amplifier receives a signal from an amplitude setting loop, wherein the signal of the amplitude setting loop is based on an input from a digital-to-analog converter and an input from a voltage sensing loop, wherein the input from the voltage sensing loop is determined using the high voltage pulse, a voltage divider and a differential amplifier.

70. The electroporation system of claim 69, wherein the common-source amplifier amplifies the signal from the amplitude setting loop by about 1,000 times to about 2,000 times.

71. An electroporation system according to claim 69 or claim 70, wherein the input from the digital to analog converter corresponds to a user selected waveform.

72. An electroporation system according to any one of claims 21 to 71, wherein the pulse generator comprises an arc detection module, which is configured to detect an arc in the sample / the sample during the application of the voltage / the voltage to the sample.

73. The electroporation system of claim 72, wherein the arc detection module include: a first stage amplifier configured to provide an amplified current signal based on a current signal associated with the application of the voltage to the sample; a bandpass filter configured to filter a falling edge signal from the amplified current signal, the falling edge signal indicating a decrease in current through the sample, the decrease in current being indicative of an arc; and A comparator is configured to compare the falling edge signal filtered by the bandpass filter with one or more reference standards to determine whether an arc has occurred in the sample.

74. The electroporation system of claim 73, wherein the amplification of the current signal by the first stage amplifier is based on an output of a low voltage detection circuit for determining a resistance associated with the sample.

75. A pulse generator, comprising one or more connection ports, each specific connection port among the one or more connection ports comprising a corresponding port door, and the corresponding port door comprising a corresponding biasing element for biasing the corresponding port door into a closed configuration to prevent access to the specific connection port.

76. A pulse generator according to claim 75, wherein the corresponding port door includes a corresponding tool interface constructed to receive a port door tool, and the port door tool is constructed to interact with the corresponding tool interface to react against the corresponding biasing element of the corresponding port door, thereby causing the corresponding port door to be in an open configuration to provide access to the specific connection port.

77. A pulse generator according to claim 76, wherein insertion of a port connection component into said specific connection port maintains said reaction on said corresponding biasing element to maintain said corresponding port door in said open configuration.

78. A pulse generator according to claim 77, wherein disconnection of the port connection component from the specific connection port eliminates the reaction on the corresponding biasing element to allow the corresponding port door to return to the closed configuration.

79. A pipette tip, the pipette tip include: a tip sleeve defining an inner cavity extending from a proximal end of the pipette tip to a distal end of the pipette tip; a plunger at least partially disposed within the lumen, the plunger being constructed of an electrically conductive material and configured to translate along the lumen to facilitate drawing a fluid into and / or dispensing a fluid from the lumen; and An attachment interface is provided at the proximal end of the pipette tip, the attachment interface comprising one or more tabs configured to engage with a distal section of a pipette.

80. The pipette tip of claim 79, wherein the pipette tip has a sample volume capacity between 10 μL and 100 μL.

81. The pipette tip of claim 80, wherein the pipette tip has a sample volume capacity of 10 μL or 100 μL.

82. The pipette tip of claim 79, wherein the plunger comprises gold, diamond-like carbon, and / or a conductive plastic.

83. A pipette tip according to claim 79, wherein the plunger includes a coupling section and an inner cavity section, and the inner cavity section includes a sealing component for creating a seal between the plunger and the inner cavity wall of the tip sleeve.

84. A pipette tip according to claim 83, wherein the inner cavity segment includes a front pin and a shaft segment, wherein the front pin is configured to connect to the shaft segment and secure the sealing component to the shaft segment.

85. A pipette tip according to claim 84, wherein the sealing component comprises a polymer sleeve, and wherein the front pin is configured to be inserted through the polymer sleeve and engage with the retaining hole of the shaft segment to secure the sealing component to the shaft segment.

86. A pipette tip according to claim 85, wherein the front pin is inserted through the polymer sleeve to define a space between the polymer sleeve and the front pin, and wherein the space contributes to the flexibility of the polymer sleeve for creating the seal between the plunger and the inner cavity.

87. A pipette tip according to any one of claims 83 to 86, wherein the sealing component comprises polytetrafluoroethylene (PTFE).

88. A pipette tip according to claim 87, wherein the sealing component comprises a coated O-ring, and wherein the inner cavity segment comprises a circumferential recess configured to receive the coated O-ring.

89. A pipette assembly, the pipette assembly include: a) a pipette, the pipette comprising: a proximal segment having a handle; a distal segment having a tip interface; and a first actuator disposed in the proximal segment; and b) a pipette tip configured to be attached to the distal section, the pipette tip comprising: a tip sleeve defining an inner cavity extending from a proximal end of the pipette tip to a distal end of the pipette tip; a plunger at least partially disposed within the lumen; and an attachment interface disposed at the proximal end of the pipette tip, wherein the plunger is reversibly operably coupled to the first actuator and, when operably coupled, performs a pipetting function upon actuation of the first actuator by translating along the lumen, and Wherein the tip sleeve is reversibly attached to the distal section via one or more tabs of the attachment interface that engage with the retention platform of the distal section.

90. The pipette assembly of claim 89, wherein the plunger is comprised of a conductive material.

91. The pipette assembly of claim 90, wherein the plunger comprises gold, diamond-like carbon, and / or a conductive plastic.

92. The pipette assembly of claim 90, further comprising a pipette electrode disposed in the distal segment, the pipette electrode being electrically coupled to the plunger when the plunger is operably coupled to the first actuator.

93. A pipette assembly according to claim 92, wherein the pipetting function includes aspirating fluid into the inner cavity or dispensing fluid from the inner cavity.

94. A pipette assembly according to claim 93, wherein the first actuator has a first undepressed position and a second partially depressed position, and in, When the plunger is operably coupled to the first actuator, transitioning the first actuator from the first undepressed position to the second partially depressed position results in dispensing, and transitioning the first actuator from the second partially depressed position to the first undepressed position results in aspiration.

95. A pipette assembly according to claim 94, wherein the first actuator has a third fully depressed position.

96. A pipette assembly according to claim 95, wherein transitioning the first actuator from the second partially depressed position to the third fully depressed position causes the plunger to be released from the pipette, and transitioning the first actuator from the third fully depressed position to the second partially depressed position causes the plunger to be retained by the pipette.

97. A pipette assembly according to claim 96, further comprising a clamp mechanism disposed in the distal segment, the clamp mechanism being operable to grasp and release the plunger when the first actuator transitions between the second partially depressed position and the third fully depressed position.

98. The pipette of claim 97, wherein the clamp mechanism include: a gripper jaw including a jaw opening for receiving and retaining an engagement segment of the plunger, the gripper jaw operably coupled to the first actuator and electrically coupled to the pipette electrode; and A clamping sleeve is positioned around the clamp jaws and is configured to apply an inward force to the clamp jaws so that the clamp jaws apply a compressive force to the engagement section of the plunger, thereby retaining the engagement section of the plunger within the clamp jaws when the first actuator is in the first depressed position or the second partially depressed position.

99. A pipette assembly according to claim 98, wherein actuation of the first actuator causes the clamp jaw to advance distally relative to the clamp sleeve to retract the inward force on the clamp jaw and release the engagement section of the plunger.

100. The pipette assembly of claim 89, further comprising a second actuator disposed in the proximal segment, the second actuator being operable, when actuated, to separate the attachment interface of the pipette tip from the retaining platform.

101. The pipette assembly of claim 100, further comprising a tip ejection sleeve operably connected to the second actuator and disposed around the holding platform, the tip ejection sleeve being operable to move distally relative to the holding platform and to separate the attachment interface of the pipette tip from the holding platform when the second actuator is actuated by transitioning the second actuator from a first undepressed position to a second depressed position.

102. The pipette assembly of claim 100, wherein the second actuator is configured to traverse a blank travel distance when depressed prior to separating the one or more tabs from the retaining platform.

103. The pipette assembly of claim 100, wherein the distal section of the pipette comprises a biasing member configured to bias the one or more tabs into engagement with the retention platform.

104. The pipette assembly of any one of claims 89 to 103, wherein the inner cavity has a sample volume capacity of between 10 μL and 100 μL.

105. A pipette assembly, the pipette assembly include: a) a pipette, the pipette comprising: a proximal segment having a handle; a distal segment having a suction tip interface; a first actuator disposed in the proximal segment; and a gripper mechanism disposed in the distal segment, the gripper mechanism having gripper jaws; and b) a pipette tip configured to be attached to the distal section, the pipette tip comprising: a tip sleeve defining an interior cavity extending from a proximal end of the pipette tip to a distal end of the pipette tip; and a plunger disposed at least partially within the lumen; and Wherein the plunger is reversibly operably coupled to the first actuator via the gripper jaws and, when operably coupled, performs a pipetting function upon actuation of the first actuator by translating along the lumen.

106. A pipette assembly according to claim 105, wherein the plunger is composed of a conductive material.

107. A pipette assembly according to claim 106, wherein the plunger comprises gold, diamond-like carbon and / or a conductive plastic.

108. The pipette assembly of claim 106, further comprising a pipette electrode disposed in the distal segment, the pipette electrode being electrically coupled to the plunger when the plunger is operably coupled to the first actuator.

109. A pipette assembly according to claim 108, wherein the pipetting function includes aspirating fluid into the inner cavity or dispensing fluid from the inner cavity.

110. The pipette assembly of claim 109, wherein the first actuator has a first, undepressed position and a second, partially depressed position, and in, When the plunger is operably coupled to the first actuator, transitioning the first actuator from the first undepressed position to the second partially depressed position results in dispensing, and transitioning the first actuator from the second partially depressed position to the first undepressed position results in aspiration.

111. The pipette assembly of claim 110, wherein the first actuator has a third fully depressed position.

112. A pipette assembly according to claim 111, wherein transitioning the first actuator from the second partially depressed position to the third fully depressed position causes release of the plunger, and transitioning the first actuator from the third fully depressed position to the second partially depressed position causes gripping of the plunger.

113. A pipette assembly according to claim 112, wherein the gripper mechanism is operable to grasp and release the plunger when the first actuator transitions between the second partially depressed position and the third fully depressed position.

114. A pipette assembly according to claim 113, wherein the clamp mechanism include: the gripper jaw, the gripper jaw comprising a jaw opening for receiving and retaining an engagement segment of the plunger, the gripper jaw operably coupled to the first actuator and electrically coupled to the pipette electrode; and A clamping sleeve is positioned around the clamp jaws and is configured to apply an inward force to the clamp jaws so that the clamp jaws apply a compressive force to the engagement section of the plunger, thereby retaining the engagement section of the plunger within the clamp jaws when the first actuator is in the first depressed position or the second partially depressed position.

115. A pipette assembly according to claim 114, wherein actuation of the first actuator causes the clamp jaw to advance distally relative to the clamp sleeve to retract the inward force on the clamp jaw and release the engagement section of the plunger.

116. The pipette assembly of claim 115, further comprising an attachment interface disposed at the proximal end of the pipette tip.

117. A pipette assembly according to claim 116, wherein the tip sleeve is reversibly attached to the distal segment via one or more tabs of the attachment interface that engage with the retention platform of the distal segment.

118. The pipette assembly of claim 117, further comprising a second actuator disposed in the proximal segment, the second actuator being operable, when actuated, to separate the attachment interface of the pipette tip from the retaining platform.

119. The pipette assembly of claim 118, further comprising a tip ejection sleeve operably connected to the second actuator and disposed around the holding platform, the tip ejection sleeve being operable to move distally relative to the holding platform and to separate the attachment interface of the pipette tip from the holding platform when the second actuator is actuated by transitioning the second actuator from a first undepressed position to a second depressed position.

120. The pipette assembly of claim 119, wherein the second actuator is configured to traverse a blank travel distance when depressed prior to decoupling the one or more tabs from the retention platform.

121. A pipette assembly according to claim 117, wherein the distal section of the pipette includes a biasing member configured to bias the one or more tabs into engagement with the retention platform.

122. The pipette assembly of any one of claims 105 to 122, wherein the inner cavity has a sample volume capacity of between 10 μL and 100 μL.

123. A method, the method comprising: include: Providing a pipette according to any one of claims 1 to 20 and a pipette tip according to any one of claims 79 to 88; contacting the attachment interface of the pipette tip with the tip interface of the pipette and coupling the one or more tabs with the holding platform to attach the tip cartridge to the pipette; transitioning the first actuator to the third fully depressed position to position the engagement segment of the plunger within the jaw opening; as well as The first actuator is transitioned to the second partially depressed position to grasp the engagement segment.

124. The method of claim 123, further comprising drawing the sample into the inner cavity by transitioning the first actuator from the second partially depressed position to the first undepressed position while the distal end of the pipette tip is in contact with the sample.

125. The method of claim 124, further comprising delivering an electric current to the sample within the lumen.

126. The method of claim 124 or claim 125, further comprising dispensing the sample from the lumen by transitioning the first actuator from the first undepressed position to the second partially depressed position.

127. The method of any one of claims 124 to 126, further comprising separating the tip sleeve from the pipette by transitioning the second actuator from the first undepressed position to the second depressed position thereby separating the one or more tabs from the retaining platform.

128. The method of claim 127, further comprising loosening the plunger by transitioning the first actuator to the third fully depressed position to release the engagement segment of the plunger from the clamp jaws.

129. A method for transfecting cells with a payload, the method include: Providing an electroporation system according to any one of claims 21 to 74; providing the cell; providing the payload; introducing the cells and the payload into the pipette tip; and electroporating the cells by operating the electroporation system.

130. The method of claim 129, wherein the cell is a mammalian cell.

131. The method of claim 129, wherein the cell is a microorganism or an organoid.

132. The method of claim 129, wherein the payload is selected from the group consisting of nucleic acids, proteins, or combinations thereof.

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