Multichannel pipette and electroporation system

Abstract

A multichannel pipette, an electroporation system utilizing a multichannel pipette, and a method for electroporating cells. The electroporation system includes a multichannel 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.

Description

Technical Field

[0001] Cross - reference to related applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 472,208, filed on June 9, 2023, the disclosure of which is considered a part of the disclosure of this application and is hereby incorporated by reference in its entirety.

[0002] The present invention generally relates to devices for fluid handling, and more specifically, to multi - channel pipettes, electroporation systems utilizing multi - channel pipettes, and methods for electroporating cells.

Background Art

[0003] Transfection involves the non - viral introduction of nucleic acids and / or proteins into cells. To facilitate transfection, various chemical or physical methods may be employed. Electroporation is a physical transfection method that uses an electrical pulse to create temporary pores in the cell membrane, allowing substances such as nucleic acids to pass into the cell.

[0004] Some electroporation systems include a pipette for holding target cells and a payload (e.g., nucleic acids and / or proteins to be introduced into the target cells) and an electrical pulse generator for providing an electrical pulse to the target cells. The pipette can be connected to or inserted into a docking station associated with the electrical pulse generator to allow the electrical pulse generated by the electrical pulse generator to reach the target cells.

[0005] For example, a pipette electrode, which is in conductive communication with one end of the pipette tip containing the liquid with target cells and payload, can interface with a first electrode on a docking station. The other end of the pipette tip can be inserted into a buffer solution (e.g., in a reservoir or buffer tube) which is in conductive communication with a second electrode on the docking station, thereby exposing the open end of the pipette tip to the buffer solution. Once the pipette and pipette tip are connected to the docking station in this manner, an electrical pulse generator can supply electrical pulses to the first and second electrodes of the docking station, thereby allowing the electrical pulses to travel through the pipette and pipette tip to reach the target cells.

[0006] Conventional electroporation systems include a single-channel pipette and are therefore limited in high-throughput applications. Thus, a drawback of conventional electroporation systems is their inability to electroporate multiple different samples simultaneously, which is desirable for high-throughput applications. The ability to process multiple samples at once significantly reduces processing time and also greatly reduces the labor required of technicians when processing large volumes of samples.

[0007] Existing electroporation systems have several drawbacks, and there is a continuing need and demand for improved electroporation systems that can be applied to high-throughput applications. [Overview of the project]

[0008] Various aspects of this disclosure extend at least to electroporation systems, their components, and / or methods associated therewith.

[0009] In one embodiment, the present disclosure provides a multichannel pipette (also referred to herein as a “pipette”). The pipette includes a proximal section having a handle; a distal section configured to reversibly engage with a plurality of pipette tips; a first actuator disposed in the proximal section; and a second actuator disposed in the proximal section and operable to control the dispensing function of the pipette. In various embodiments, the first actuator is operable to control aspiration, and the second actuator is operable to control dispensing.

[0010] In various embodiments, the pipette also further includes a third actuator disposed in the proximal section, the third actuator configured to disengage the pipette tip attached to the distal section of the pipette when the third actuator is actuated. In some embodiments, the first actuator is configured to move from a first unpressed position to a second pressed position, and the second actuator is configured to move from the first pressed position to a second unpressed position when the first actuator moves from the first unpressed position to the second pressed position, thereby producing an aspiration function by moving the first actuator from the first unpressed position to the second pressed position, and producing a dispensing function by moving the second actuator from the second unpressed position to the first pressed position.

[0011] In various embodiments, the pipette includes a plurality of gripper mechanisms disposed in the distal section, each gripper mechanism configured to reversibly grip a plunger disposed in the lumen of the pipette tip, the plunger having an engagement section for engaging with the gripper mechanism and a lumen section disposed in the lumen of the pipette tip. In some embodiments, the pipette includes a lock button disposed in the proximal section, which is operably connected to a locking mechanism configured to control the locking of multiple gripper mechanisms. When the lock button is pressed, the locking mechanism transitions from a first unlocked configuration to a second locked configuration. In addition, when the locking mechanism is in the first configuration, the multiple gripper mechanisms are open and configured to receive the engaging sections of the plungers, and when the locking mechanism is in the second configuration, the multiple gripper mechanisms are closed and configured to grip and engage with the engaging sections of the plungers.

[0012] In another embodiment, the Disclosure provides an electroporation system comprising 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.

[0013] In yet another embodiment, the Disclosure provides a method for transfecting cells with a payload. This method includes providing an electroporation system of the Disclosure, providing cells, providing a payload, introducing the cells and payload into the tip of a pipette, and electroporating the cells by operating the electroporation system. In some embodiments, the cells are mammalian cells. In some embodiments, the payload includes nucleic acids, proteins, or combinations thereof.

[0014] To gain a more complete understanding of the nature and merits of the present invention, refer to the following detailed description taken in conjunction with the accompanying drawings. It should be understood that these drawings represent only representative embodiments of the present invention and are therefore not to be considered limiting its scope. This disclosure is described and explained with additional specificities and details using the accompanying drawings. [Brief explanation of the drawing]

[0015] [Figure 1] These are exemplary components of the electroporation system of the present disclosure, and schematic diagrams illustrating such components in one embodiment of the present disclosure. [Figure 2] These are front, right-side, and elevation perspective views of one embodiment of the multichannel pipette disclosed herein. [Figure 3] Figure 2 is a right-side view of the multichannel pipette depicted. [Figure 4] Figure 2 is a left side view of the multichannel pipette depicted. [Figure 5] Figure 2 shows a rear view of the multichannel pipette. [Figure 6] Figure 2 is a front view of the multichannel pipette. [Figure 7] Figure 2 is a top view of the multichannel pipette. [Figure 8] Figure 2 is a bottom view of the multichannel pipette. [Figure 9] This is a front, right-side, and elevation perspective view of one embodiment of the multichannel pipette of the present disclosure, the pipette having a pipette tip attached to the distal section of the pipette. [Figure 10] Figure 9 is a right-side view of the multichannel pipette depicted. [Figure 11] Figure 9 is a left side view of the multichannel pipette depicted. [Figure 12] Figure 9 shows a rear view of the multichannel pipette. [Figure 13] Figure 9 is a front view of the multichannel pipette. [Figure 14] Figure 9 is a top view of the multichannel pipette. [Figure 15] Figure 9 is a bottom view of the multichannel pipette depicted. [Figure 16A]A schematic diagram illustrating aspects of an exemplary pipette tip (e.g., a consumable pipette tip) for use with an electroporation system in an embodiment of the present disclosure. FIG. 16A is a schematic diagram illustrating aspects of an exemplary pipette tip in an embodiment of the present disclosure. [Figure 16B] A schematic diagram illustrating aspects of an exemplary pipette tip (e.g., a consumable pipette tip) for use with an electroporation system in an embodiment of the present disclosure. FIG. 16B is a schematic diagram illustrating aspects of an exemplary pipette tip in an embodiment of the present disclosure. [Figure 17A] A schematic diagram showing an exploded view of the pipette tip depicted in FIGS. 16A and 16B. FIG. 17A is an exploded view of the pipette tip depicted in FIG. 16A. [Figure 17B] A schematic diagram showing an exploded view of the pipette tip depicted in FIGS. 16A and 16B. FIG. 17B is an exploded view of the pipette tip depicted in FIG. 16B. [Figure 18A] A schematic diagram illustrating aspects of a plunger of a pipette tip in an embodiment of the present disclosure. FIG. 18A is an exploded view of an exemplary plunger. [Figure 18B] A schematic diagram illustrating aspects of a plunger of a pipette tip in an embodiment of the present disclosure. FIG. 18B is a schematic diagram of the assembled plunger depicted in FIG. 18A. [Figure 19A] A schematic diagram illustrating aspects of a pipette tip in an embodiment of the present disclosure. FIG. 19A is a cross-sectional view of a pipette tip in an embodiment of the present disclosure. [Figure 19B] A schematic diagram illustrating aspects of a pipette tip in an embodiment of the present disclosure. FIG. 19B is an enlarged cross-sectional view of the distal portion of the pipette tip depicted in FIG. 19A. [Figure 20] A schematic diagram illustrating aspects of a plunger of a pipette tip in an embodiment of the present disclosure. [Figure 21A] A schematic diagram illustrating a pipette tip in an embodiment of the present disclosure. FIG. 21A is a schematic diagram of the pipette tip of the present disclosure. [Figure 21B]Figure 21B is a schematic diagram illustrating the tip of a pipette in one embodiment of the present disclosure, and is a magnified view of the distal portion of the pipette tip depicted in Figure 21A. [Figure 22] This is a perspective view of a pipette tip for use with a multichannel pipette in an embodiment of the present disclosure. [Figure 23] This is an elevation perspective view of a multichannel pipette in one embodiment of the present disclosure. [Figure 24] Figure 23 is a cross-sectional side view of the pipette. [Figure 25] The function of the internal mechanism of the pipette shown in Figure 23 is illustrated by illustrating how the second actuator moves to the first pressed position to dispense fluid. [Figure 26] The function of the internal mechanism of the pipette shown in Figure 23 is illustrated by illustrating how the first actuator moves to the second pressed position to draw in fluid. [Figure 27] The positioning of internal components of a pipette is illustrated when the second actuator is in the first pressed position after the dispensing function has been performed, or when the pipette is configured to lock and secure the plunger at the attached pipette tip by pressing the lock button. [Figure 28] This illustrates the internal components of a pipette that operably connect a lock button to a gripping sleeve via a push plate. [Figure 29] The positioning of the internal components of the pipette after the lock button is pressed and the second actuator is fully depressed is illustrated. It is shown that the plungers at the tips of each pipette are gripped and locked via individual gripper mechanisms. [Figure 30] Figure 28 shows a magnified view of the rectangular area enclosed by the dashed line, which illustrates the plunger at the tip of the pipette being locked and gripped by the gripper jaws of the gripper mechanism. The gripping sleeve moves distally relative to its distal end, closing the gripper jaws and locking the plunger at the tip of the pipette with the gripper mechanism. [Figure 31] This is an elevation perspective view of a multichannel pipette in one embodiment of the present disclosure. [Figure 32] The function of the internal mechanism of the pipette shown in Figure 31 is illustrated by the second actuator moving to the first pressed position to dispense the fluid. [Figure 33] The function of the internal mechanism of the pipette shown in Figure 31 is illustrated by illustrating how the first actuator moves to the second pressed position to draw in fluid. [Figure 34] The positioning of internal components of a pipette is illustrated when the second actuator is in the first pressed position after the dispensing function has been performed, or when the pipette is configured to lock and secure the plunger at the attached pipette tip by pressing the lock button. [Figure 35] The positioning of the internal components of the pipette after the lock button is pressed and the second actuator is fully depressed is illustrated. It is shown that the plungers at the tips of each pipette are gripped and locked via individual gripper mechanisms. [Figure 36] Figure 35 is an enlarged view of the rectangular area enclosed by the dashed line, showing the plunger at the tip of the pipette locked and gripped by the gripper jaws of the gripper mechanism. The gripping sleeve moves distally to the distal tip of the gripper jaws, closing the gripper jaws and gripping and locking the plunger at the tip of the pipette with the gripper mechanism. [Figure 37] This schematic diagram illustrates an open-configuration gripper mechanism in an embodiment of the present disclosure, such that the engaging section of the pipette tip plunger can be received by the gripper jaws. [Figure 38] Figure 37 is a schematic diagram illustrating an open gripper mechanism in an embodiment of the present disclosure, so that the engaging section of the pipette tip plunger can be received by the gripper jaws. [Figure 39] Figure 37 is a schematic diagram illustrating an open gripper mechanism in an embodiment of the present disclosure, so that the engaging section of the pipette tip plunger can be received by the gripper jaws. [Figure 40]This schematic diagram illustrates a closed gripper mechanism in an embodiment of the present disclosure, such that the engaging section of the pipette tip plunger can be gripped by the gripper jaws. [Figure 41] Figure 40 is a schematic diagram illustrating a closed gripper mechanism in an embodiment of the present disclosure, so that the engaging section of the pipette tip plunger can be gripped by the gripper jaws. [Figure 42] This is a schematic diagram illustrating the arrangement and interaction between a magnet disposed on the outer housing of a multichannel pipette and a linkage bar disposed inside, in an embodiment of the present disclosure. [Figure 43] This is a schematic diagram illustrating the arrangement and interaction between a magnet disposed on the outer housing of a multichannel pipette and a linkage bar disposed inside, in an embodiment of the present disclosure. [Figure 44] This is a schematic diagram illustrating the arrangement and interaction between a magnet disposed on the outer housing of a multichannel pipette and a linkage bar disposed inside, in an embodiment of the present disclosure. [Figure 45] Figure 31 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 46] This is a schematic diagram showing the linkage bar when the second actuator is in the first pressed position in an embodiment of the present disclosure, and the pipette mis-trigger prevention shown in Figure 31. [Figure 47] This is a schematic diagram illustrating the step of disengaging the pipette tip after an accidental trigger run of the pipette shown in Figure 31, according to an embodiment of the present disclosure. [Figure 48] This schematic diagram illustrates a step of disengaging the pipette tip in an embodiment of the present disclosure, which involves simultaneously disengaging the plunger shown in Figure 31 and releasing the pipette, as well as disengaging and releasing the pipette tip attachment interface. [Figure 49] Figure 31 is a schematic diagram illustrating the functionality of the pipette tip discharge sleeve shown in the embodiment of this disclosure. [Figure 50] This is a schematic diagram illustrating the internal components of a pipette used to reduce the force during disengagement of the pipette tip. The force reduction is achieved using a lever configuration between the fulcrum and the load. [Figure 51] Figure 31 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 52] This is a schematic diagram showing the linkage bar when the second actuator is in the first pressed position in an embodiment of the present disclosure, and the pipette mis-trigger prevention shown in Figure 31. [Figure 53] This schematic diagram illustrates partial tip ejection in an embodiment of the present disclosure, where the third actuator of the pipette shown in Figure 31 is moved to a partially depressed position until it is fully depressed, causing the linkage bar to move distally, but the pipette tip plunger remains engaged. [Figure 54] Figure 23 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 55] Figure 23 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 56] Figure 23 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 57] Figure 23 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 58] Figure 23 is a schematic diagram illustrating the internal mechanism and function of the pipette shown in the embodiments of this disclosure, which is used to displace and remove the attached pipette tip. [Figure 59]This figure illustrates an electroporation system in one embodiment of the present disclosure. [Figure 60] Figure 59 is an elevation front perspective view of the pipette docking assembly shown in the embodiment of the present disclosure, with the pipette and reservoir not docked. [Figure 61] Figure 59 is an elevation and rear perspective view of the pipette docking assembly shown in the embodiment of the present disclosure, with the pipette and reservoir not docked. [Figure 62] This is a schematic diagram illustrating the components of the reservoir shown in Figure 59 in an embodiment of the present disclosure. [Figure 63] This is a schematic diagram illustrating an example of a pipette station guard in an embodiment of the present disclosure. [Figure 64] Figure 63 is a schematic diagram illustrating the connection of the pipette station guard to the pipette docking station. [Figure 65] Figure 63 is a schematic diagram illustrating the connection of the pipette station guard to the pipette docking station. [Figure 66] This is a schematic diagram illustrating an alternative gear configuration for the gear mechanism of the pipette shown in Figure 32 in an embodiment of the present disclosure. [Figure 67] This is an elevation perspective view of a multichannel pipette in one embodiment of the present disclosure. [Figure 68] This is an elevation perspective view of the multichannel pipette of the present disclosure, which engages with the pipette tip. [Figure 69] Figure 67 illustrates the function of the internal mechanism of the pipette when the lock button is pressed. [Figure 70] The function of the internal mechanism of the pipette shown in Figure 67 is illustrated by the first actuator moving to the second pressed position to draw in fluid. [Figure 71] Figure 67 illustrates the positioning of the internal components of the pipette before the discharge button is pressed. [Figure 72]Figure 67 illustrates the internal components of the pipette, which are operably connected to the pipette tip when the lock button is pressed and the plunger is gripped by gripping the jaws. [Figure 73] Let's illustrate this with an example using the gripper jaw of the pipette shown in Figure 67, which is in the closed position. [Figure 74] The function of the internal mechanism of the pipette in Figure 67 is illustrated when the first actuator is moved to a second depressed position after the tip has been loaded into one embodiment of the present disclosure. [Figure 75] Let us illustrate with an example the pipette gripper jaw shown in Figure 74 when the first actuator is moved to the second pressed position. [Figure 76] Figure 67 illustrates the tip loaded into the pipette. [Figure 77] Figure 67 illustrates an example of the gripper mechanism of a pipette. [Figure 78] Figure 67 is a cross-sectional view illustrating an example of the gripper mechanism of a pipette. [Figure 79] Figure 67 illustrates an example of the gripper mechanism of a pipette. [Figure 80] Figure 67 is a cross-sectional view illustrating an example of the gripper mechanism of a pipette. [Figure 81] Figure 67 illustrates an example of a pipette push plate assembly. [Figure 82] Figure 67 illustrates the function of the internal mechanism of the pipette when the lock button is deactivated. [Figure 83] With the gripper jaws open, we illustrate the configuration of the pipette gripper mechanism shown in Figure 67. [Figure 84] Figure 67 illustrates the function of the internal mechanism of the pipette when the lock button is activated, for example, when it is pressed. [Figure 85] Figure 67 illustrates the configuration of the pipette's gripper mechanism with the gripper jaw closed. [Figure 86]Figure 67 illustrates the function of the internal mechanism of the pipette when the lock button is activated, for example, when it is pressed. [Figure 87] The function of the internal mechanism of the pipette shown in Figure 67 is illustrated by the first actuator moving to the pressed position to draw in fluid. [Figure 88] The function of the internal mechanism of the pipette shown in Figure 67 is illustrated by the first actuator moving to the pressed position to dispense fluid. [Figure 89] Figure 67 illustrates the positioning of the internal mechanism of the pipette when the first actuator moves to the pressed position and draws in fluid. [Figure 90] Figure 67 illustrates the positioning of the internal components of the pipette before the discharge button is pressed. [Figure 91] Figure 67 illustrates an example of a pipette push plate assembly. [Figure 92] An example illustrates the internal configuration of a pipette station top assembly having anode and cathode modules for electroporation. [Figure 93] An example illustrates the internal configuration of a pipette station top assembly having anode and cathode modules for electroporation. [Figure 94] The internal configuration of the pipette station bottom assembly is illustrated with an example. [Figure 95] The internal configuration of the pipette station bottom assembly is illustrated with an example. [Figure 96] This example illustrates the configuration of the cathode module in a pipette station top assembly. [Figure 97] This example illustrates the configuration of the cathode module in a pipette station top assembly. [Figure 98] This document illustrates an example of the anode module of a pipette station top assembly. [Figure 99] This document illustrates an example of the anode module of a pipette station top assembly. [Figure 100]Exemplary components of the electroporation system of this disclosure in several embodiments are illustrated below. [Figure 101] This is a front right-side perspective view of one embodiment of a docking assembly with a pipette docked. [Figure 102] This is a front right-side perspective view of the docking assembly. [Figure 103] This is a front right-side perspective view of the pipette station. [Figure 104] Figure 103 is a front view of the pipette station. [Figure 105] Figure 103 is a rear view of the pipette station. [Figure 106] Figure 103 is a right-side elevation view of the pipette station. [Figure 107] Figure 103 is a left side view of the pipette station. [Figure 108] Figure 103 is a top view of the pipette station. [Figure 109] Figure 103 is a bottom view of the pipette station. [Figure 110] This is a front right-side perspective view of the station guard. [Figure 111] Figure 110 is a front view of the station guard. [Figure 112] Figure 110 is a rear view of the station guard. [Figure 113] Figure 110 is a right-side elevation view of the station guard. [Figure 114] Figure 110 is a left side view of the station guard. [Figure 115] Figure 110 is a top view of the station guard. [Figure 116] Figure 110 is a bottom view of the station guard. [Figure 117] This is a front right-side perspective view of the reservoir. [Figure 118] Figure 117 is a front view of the reservoir. [Figure 119] Figure 117 is a rear view of the reservoir. [Figure 120]Figure 117 is a right-side elevation view of the reservoir. [Figure 121] Figure 117 is a left side view of the reservoir. [Figure 122] Figure 117 is a top view of the reservoir. [Figure 123] Figure 117 is a bottom view of the reservoir. [Figure 124] This is a front-right perspective view of a multichannel pipette. [Figure 125] Figure 124 is a front view of a multichannel pipette. [Figure 126] Figure 124 is a rear view of the multichannel pipette. [Figure 127] Figure 124 is a right-side elevation view of a multichannel pipette. [Figure 128] Figure 124 is a left side view of a multichannel pipette. [Figure 129] Figure 124 is a top view of a multichannel pipette. [Figure 130] Figure 124 is a bottom view of a multichannel pipette. [Figure 131] This is a front-right perspective view of a multichannel pipette with the pipette tip attached. [Figure 132] Figure 131 is a front view of a multichannel pipette. [Figure 133] Figure 131 is a rear view of the multichannel pipette. [Figure 134] Figure 131 is a right-side elevation view of a multichannel pipette. [Figure 135] Figure 131 is a left side view of a multichannel pipette. [Figure 136] Figure 131 is a top view of a multichannel pipette. [Figure 137] Figure 131 is a bottom view of a multichannel pipette. [Figure 138] This is a front right-side perspective view of a pipette tip plunger in one embodiment of the present disclosure. [Figure 139] Figure 138 is a front view of the pipette tip plunger. [Modes for carrying out the invention]

[0016] Embodiments of this disclosure extend at least to pipettes used in electroporation (e.g., multichannel pipettes), and to electroporation systems and / or components thereof that utilize such pipettes. The disclosed embodiments and designs may be implemented to address various shortcomings associated with at least some conventional pipettes and electroporation systems and / or techniques. The following discussion outlines some exemplary improvements and / or practical applications that may 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 exemplary improvements discussed herein.

[0017] Some embodiments of this disclosure provide pipettes that enable the simultaneous processing of multiple samples. The unique design of the multichannel pipettes described herein reduces user muscle stress and / or fatigue by reducing the force involved in manually performing the pipetting function on a device that processes multiple samples simultaneously using multiple pipette tips. In addition, the pipettes of this disclosure are designed to utilize pipette tips through which a clip-on connection is achieved between the pipette tip and the pipette. By using a clip-on connection along with reducing the force that contributes to muscle stress and / or fatigue during pipetting, the pipettes and systems described herein are ideal for high-throughput applications.

[0018] Before describing in detail the various embodiments of this disclosure, it should be understood that this disclosure is not limited to the parameters of the systems, methods, apparatus, products, processes, consumables, and / or kits, which are naturally subject to change. Therefore, while certain embodiments of this disclosure are described in detail with reference to specific configurations, parameters, components, elements, etc., the descriptions are illustrative and should not be construed as limiting the scope of the claimed invention. Furthermore, the terms used herein are intended to describe embodiments and are not necessarily intended to limit the scope of the claimed invention.

[0019] Furthermore, unless otherwise implicitly or explicitly understood or stated, it is understood that, with respect to any given component or embodiment described herein, any of the possible candidates or substitutes listed for that component may generally be used individually or in combination with each other. In addition, unless otherwise implicitly or explicitly understood or stated, it will be understood that any such list of candidates or substitutes is merely illustrative and not limiting.

[0020] In addition, unless otherwise indicated, numbers representing quantities, components, distances, or other measurements used in the specification and claims should be understood as being modified by the term “about” as defined herein. Accordingly, unless otherwise indicated, numerical parameters described in the specification and appended claims are approximations that may vary depending on the desired characteristics to be obtained from the subject matter presented herein. At a minimum, and without intending to limit the application of the principle of equivalents to the claims, each numerical parameter should be interpreted at least in light of the number of significant figures reported and by applying ordinary rounding techniques. Although numerical ranges and parameters describing a broad range of the subject matter presented herein are approximations, the figures described in specific examples are reported as accurately as possible. However, any figure inherently contains a certain degree of error that inevitably arises from the standard deviation observed in each experimental measurement.

[0021] The term "comprising," which is synonymous with "including," "containing," "having," or "characterized by," is comprehensive or open-ended and does not exclude additional, unincorporated elements or methodological steps.

[0022] Note that, as used herein and in the appended claims, the singular forms "a," "an," and "the" refer to multiple objects unless explicitly indicated otherwise. Therefore, for example, a reference to "port" may include one, two, or more ports.

[0023] As used herein and in the appended claims, directional terms such as “top,” “bottom,” “left,” “right,” “up,” “down,” “upper,” “lower,” “inner,” “outer,” “internal,” “external,” “interior,” “exterior,” “proximal,” and “distal” are used herein solely to indicate relative directions and are not intended to limit the scope of this disclosure or the claims in any other respect.

[0024] Where possible, similar numbering of elements is used in various diagrams. Furthermore, each alternative configuration of a particular element may include a separate character attached to the element number. Thus, the attached character may be used to design alternative designs, structures, functions, implementations, and / or embodiments of elements or features that do not have the attached character. For example, the element "80" may be embodied in a different configuration and designated as "80a". Similarly, multiple instances of an element and / or a sub-element of a parent element may each include a separate character attached to the element number. In each case, the element label may generally be used without the attached character to refer to any one of all instances or alternative elements of the element. Element labels with attached characters may be used to refer to a particular instance of the element, or to distinguish or draw attention to multiple uses of that element.

[0025] Various aspects of the devices, systems, and methods of the present invention may be illustrated by reference to one or more exemplary embodiments. As used herein, the term “embodiment” means “serving as an example, illustration, or demonstrative” and should not necessarily be construed as being more preferable or advantageous than other embodiments disclosed herein.

[0026] Various aspects of this device and system can be illustrated by describing components that are coupled, attached, and / or joined together. As used herein, the terms “coupled,” “attached,” “connected,” and / or “joined” are used to indicate a direct connection between two components, or, as appropriate, an indirect connection between them via an intervening or intermediate component. In contrast, when one component is referred to as “directly coupled,” “directly attached,” “directly connected,” and / or “directly joined” to another component, there is no intervening element.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to whom this disclosure relates. Many similar or equivalent methods and materials can be used in carrying out this disclosure, but preferred materials and methods are described herein.

[0028] Electroporation system.

[0029] Figure 1 illustrates various components of an electroporation system 10 that may be used to carry out one or more disclosed embodiments. For example, the electroporation system 10 in Figure 1 may be configured to facilitate cell transfection by applying an electric current to target cells to introduce a payload into the target cells. While Figure 1 illustrates an electroporation system 10 with specific components, in consideration of this disclosure, it will be understood that the electroporation system 10 may include any number of additional and / or alternative components. Furthermore, in consideration of this disclosure, it will be understood that the principles disclosed herein are not limited to the specific form and / or features of the electroporation system 10 shown in Figure 1.

[0030] Figure 1 illustrates an electroporation system 10 including a processor 102, storage 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, including 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 104. The storage 104 may include physical system memory, which may be volatile, non-volatile, or a combination thereof. Furthermore, the storage 104 may include local storage, remote storage (e.g., accessible via the communication system 112 or otherwise), or a combination thereof. Additional details relating to the processor (e.g., processor 102), computer storage media (e.g., storage 104), and other computer components are provided below.

[0031] The processor 102 may be configured to execute instructions 106 stored in the storage 104 to perform a specific action and / or command (e.g., voltage / current control, user interface presentation, user input reception, component detection, etc.). The action may depend at least in part on data 108 stored in the storage 104 in a volatile or non-volatile manner.

[0032] In some examples, the action may rely at least in part on a communication system 112 for receiving data from a remote system 114, which may include, for example, computing devices, sensors, and / or other things. The communication system 112 may include any combination of software or hardware components that are operable to facilitate communication between on-system components / devices and / or with off-system components / devices. For example, the communication system 112 may include ports, buses, or other physical connection devices for communicating with other devices / components. In addition, or alternatively, the communication system 112 may, in non-limiting embodiments, include systems / components that are operable to communicate wirelessly with external systems and / or devices through any suitable communication channel, such as Bluetooth, ultra-wideband, WLAN, infrared communication, and / or other things.

[0033] Furthermore, Figure 1 illustrates that the electroporation system 10 may include or communicate with an I / O system 110. The I / O system 110 may include, but not limited to, any type of input or output device, such as a display, touchscreen, mouse, keyboard or button interface, controller, and / or others, in non-limiting embodiments. For example, Figure 1 illustrates that the electroporation system 10 includes a user interface element implemented in the form of a graphical touchscreen user interface on a pulse generator 100. The user interface element is configured to display information related to the operation of the electroporation system 10 and / or to receive user input to facilitate control of the electroporation system 10 (e.g., selecting, starting, monitoring, optimizing, and / or ending parameters for the electroporation process).

[0034] In various embodiments, the system includes one or more machine learning modules. Such modules may be used to optimize parameters of one or more embodiments of the system described herein, for example, to generate and / or optimize protocols, data storage, event / alarm recording, batch control, functional operation between instruments, and the like.

[0035] In certain embodiments, the Disclosure provides a system for electroporation comprising a computing device (e.g., a remote server or otherwise electronically communicating with a system) having one or more processors and memory, the memory storing instructions that, when executed by the one or more processors, cause the computing device to perform one of the methods disclosed herein. For example, the memory may store one or more weights associated with one or more trained machine learning models, including one or more trained neural networks, as disclosed herein. The term “weights” as used herein in reference to a neural network refers to all parameter values ​​and network structure definitions necessary to propagate input data through the neural network to obtain output values.

[0036] The system embodiments disclosed herein can achieve improved performance compared to conventional approaches. Therefore, in these embodiments, the present invention utilizes machine learning models, such as neural networks, in combination with an integrated high-speed computing architecture, to enable improved instrument operation, either individually or as a set of instruments and / or as a system.

[0037] Various methods and systems of embodiments disclosed herein can improve upon conventional approaches to achieve technical advantages of higher throughput, more accurate algorithms, and faster, more robust processing by utilizing machine learning models, such as trained neural networks that enable real-time processing of data, and representations of bioprocessing data. Such technical advantages cannot be achieved by routine and conventional approaches, and all users of systems including such embodiments may benefit from these advantages by assisting users in performing technical tasks such as real-time high-throughput processing, for example, through guided human-machine interaction processes. Accordingly, the technical features of embodiments disclosed herein are clearly novel in the field of filling and finishing equipment, as they are a combination of the features of embodiments disclosed herein. Thus, this disclosure introduces functionality that neither conventional computing devices nor humans have been able to perform. As used herein, the term “weights” in relation to a neural network refers to the definition of all parameter values ​​and network structure necessary to propagate input data through the neural network to obtain output values.

[0038] The electroporation system 10 includes various physical components that can be used to facilitate the electroporation operation. For example, Figures 1 and 100 illustrate that the electroporation system 10 includes a pulse generator 100 configured to supply electrical pulses to other components of the electroporation system 10. The pulse generator 100 may supply electrical pulses via a cable 105, which may selectively connect the pulse generator 100 to one or more other components of the electroporation system 10. For example, the cable 105 may connect the pulse generator 100 to a pipette docking assembly 110 to supply current to target cells present in the pipette tip of a multichannel pipette 130 connected to the pipette docking assembly 110. The pipette docking assembly 110 may include a pipette docking station 115 (to which the cable 105 may be connected), a pipette station guard 120, and a reservoir 125 configured to hold an electrolytic buffer and receive the distal section of the pipette 130, as well as the pipette tip attached thereto. Additional embodiments of the components of the electroporation system 10 are described in more detail below.

[0039] pipette tip

[0040] Figures 16–21 illustrate embodiments of pipette tips 200 for use with the multichannel pipette 130 and electroporation system 10 of the present disclosure. Such pipette tips may be attached to a pipette (e.g., pipette 130) and may hold cells and payloads to facilitate electroporation. Figure 16 depicts embodiments of 10 μL pipette tip 202 and 100 μL pipette tip 204. Although only the 10 μL and 100 μL sizes are shown in Figure 16, other sizes are also within the scope of the present disclosure.

[0041] Figure 17 shows that a 100 μL pipette tip 204 may include a plunger 302 configured to be at least partially positioned within the lumen 304 (Figure 16 shows the plunger 302 fully inserted into the lumen 304). The plunger 302 is configured to translate along the lumen 304 to facilitate pipetting functions (e.g., aspiration and / or dispensing). For example, the open end 306 of the lumen 304 may be positioned within a reservoir containing cells and a payload (e.g., nucleic acids, proteins, etc.), and the plunger 302 may be retracted from the open end 306 of the lumen 304 to draw the cells and payload into the lumen 304. The 100 μL pipette tip 204 may then be connected to other components of an electroporation system (e.g., the pipette docking assembly 110 in Figure 1) to electroporate cells to introduce a payload into them.

[0042] At least a portion of the plunger 302 may include a conductive material to allow electrical pulses to reach and / or pass through the contents of the lumen 304. For example, the plunger 302 may be coated with, formed from, or otherwise comprise gold (e.g., gold plating), diamond-like carbon, conductive plastic, and / or any other conductive medical-grade material (e.g., a material inert to mammalian cells).

[0043] Figure 17 also shows that a 10 μL pipette tip 202 may include a plunger 310 and a lumen 312 (with an open end 314), similar to the plunger 302 and lumen 304 of a 100 μL pipette tip 204.

[0044] In conventional pipette tips, the seal is created by the plunger and lumen by a metal ring on the plunger that interfaces with the inner surface of the lumen. The amount of frictional force between the metal ring and the lumen can affect the pushing / pulling force required to operate the pipette. The amount of frictional force between the metal ring and the lumen can be affected by the amount of interference between the metal ring and the lumen. As an exemplary example, for a 10 μL tip, interference in the range of 0–30 μm can produce a pushing / pulling force in the range of 0–6 N to operate the pipette. For larger tips, such as a 10 μL tip, interference in the range of 0–10 μm can produce a pushing / pulling force in the range of 0–6 N to operate the pipette. It can be difficult to consistently and reliably achieve interference within the range of 0–10 μm during production, which can result in pipette tips (especially larger pipette tips) with excessive interference between the metal ring and the lumen, leading to excessive pushing / pulling forces (e.g., exceeding 6 N) required to operate the pipette.

[0045] Therefore, at least some of the pipette tips 200 of this disclosure may implement alternative sealing components to create a seal between the lumen and the plunger. This may be particularly beneficial for larger sized pipette tips (e.g., 100 μL pipette tips).

[0046] Figure 18 illustrates an exemplary plunger 402 for a 100 μL pipette tip (in both disassembled and assembled configurations). In the embodiment of Figure 18, the plunger 402 includes an engagement section 404 and a lumen section 406. The engagement section 404 is configured to engage with a portion of the gripper mechanism (e.g., gripper jaws) of the multichannel pipette 130, as will be described in more detail below. The lumen section 406 is positioned within the lumen of the pipette tip and is configured to translate along the lumen of the pipette tip.

[0047] As shown in Figure 18, the lumen section includes a sealing component 410 that creates a seal between the plunger 402 and the lumen in which the plunger is positioned. Figure 19 shows a cross-sectional view of the lumen section 406 of the plunger 402 positioned within the lumen 502. Figure 19 depicts the interference region between the sealing component 410 and the lumen 502. Advantageously, the interference region does not extend along the entire length of the lumen section 406 of the plunger 402 positioned within the lumen 502, thereby facilitating a reduction in frictional forces between the plunger 402 and the lumen 502.

[0048] In the embodiments shown in Figures 18 and 19, the seal component 410 is implemented as a polymer sleeve (other forms are possible, such as the O-ring design shown in Figures 20 and 21). The seal component 410 may include various types of materials, such as polytetrafluoroethylene (PTFE), other Teflon materials, and / or other flexible and biocompatible materials.

[0049] The seal component 410 can be attached to the lumen section 406 of the plunger 402 in various ways. In the embodiments shown in Figures 18 and 19, the lumen section includes a front pin 412 and a shaft section 414. The front pin 412 is configured to connect to the shaft section 414, for example, by inserting a portion of the front pin 412 into a retaining hole 416 of the shaft section 414. The seal component 410 can be further secured to the shaft section 414 by inserting the front pin 412 through an opening in the seal component 410 before the front pin 412 enters the retaining hole 416 of the shaft section 414.

[0050] In some examples, when the seal component 410 is fixed to the lumen section 406, a space is formed between at least a portion of the seal component 410 and at least a portion of the lumen section 406. This can contribute to the flexibility of the seal component 410 in creating a seal between the lumen section 406 and the lumen 502 (for example, reducing frictional forces between them while still maintaining the seal). Figure 19 illustrates the space 504 formed between the seal component 410 and the front pin 412 when the front pin 412 is inserted through the seal component 410.

[0051] In light of this disclosure, it will be understood that other methods for securing the seal component 410 to the lumen section 406 (e.g., adhesives, mechanical fittings, screw connections, etc.) may be implemented in accordance with this disclosure.

[0052] As described above, the sealing component of a plunger can take various forms. Figures 20 and 21 show alternative forms of the sealing component. Figure 20 illustrates a plunger 602 in which the sealing component is implemented as an O-ring 604, which may be coated (e.g., with an inert lubricating material). The lumen section 606 of the plunger 602 includes a circumferential recess 608 configured to receive the O-ring 604. Figure 21 shows the O-ring 604 interfaced with the lumen 702 to form a seal between the lumen section 606 and the lumen 702. In some examples, the O-ring design may require greater force than the polymer sleeve design to facilitate the seal between the plunger and the pipette lumen (e.g., considering the lack of internal space in the O-ring design).

[0053] Figures 138-139 show an example of a pipette tip plunger having a tapered engagement section to improve interaction with the pipette's gripping jaws.

[0054] Multichannel pipette

[0055] Figures 2–15 and 124–137 illustrate a multichannel pipette 130 used in system 10 of the present disclosure. While the present disclosure illustrates an embodiment of the pipette 130 configured to connect with up to eight pipette tips for processing samples, it will be understood that the pipette 130 of the present disclosure may be configured to connect with two, three, four, five, six, seven, eight, nine, ten or more pipette tips 200 while maintaining the same functionality. In the embodiment, the multichannel pipette 130 includes a proximal section 135 having a handle 140, a distal section 145 configured to reversibly engage with a plurality of pipette tips 200, a first actuator 150 disposed in the proximal section 135, and a second actuator 155 disposed in the proximal section 135 and operable to control the dispensing function of the multichannel pipette 130. In various embodiments, a first actuator 150 is operable to control aspiration, and a second actuator 155 is operable to control dispensing. In embodiments, the pipette further includes a third actuator 160 located in the proximal section 135, configured to disengage a pipette tip 200 attached to the distal section of the multichannel pipette 130 when the third actuator 160 is actuated, as well as a lock button 165 located in the proximal section 135. It will be understood that the actuators (150, 155, and 160) and the lock button 165 are oriented so that the operation of each of their respective functions is controllable by the thumb of the user gripping the handle 140 and operating the pipette 130. The pipette 130 also includes a pipette electrode 132 that contacts an electrode located on the docking station when the pipette is docked to the pipette docking assembly (see Figure 3).

[0056] In various embodiments, during the operation of the pipette 130, a first actuator 150 controls the aspiration of fluid to the pipette tip 200 attached to the distal section 145 of the pipette 130. A second actuator 155 controls the dispensing of fluid from the attached pipette tip 200. A lock button 165 functions to activate a locking mechanism located in the distal section 145 of the pipette to grip and lock the plunger lumenically located within each pipette tip 200 during an electroporation protocol, including sample aspiration, electroporation, and dispensing. A third actuator 160 functions to disengage the locking mechanism and simultaneously remove the pipette tip 200 attached via a clip-on connection from the pipette 130, as further discussed herein.

[0057] As described above, one or more pipette tips can be selectively attached to pipette 130 via clip-on connections. Figure 22 illustrates a pipette tip 802 corresponding to the pipette tips shown in Figures 16 and 17, which includes features enabling a clip-on connection with the pipette. As illustrated in Figure 22, the clip-on connection includes a mounting interface 806 adjacent to the lumen 804 of the pipette tip 802. The mounting interface 806 includes tabs 808 (any number of tabs may be available) configured to engage with the corresponding mounting features of pipette 130. The tabs may be angled toward the lumen of pipette tip 802. The corresponding mounting features of pipette 130 may be located in the distal section 145 of pipette 130 for individual gripper mechanisms, which will be discussed in more detail herein.

[0058] The operation of the pipette 130 for aspirating, dispensing, and locking the pipette tip is shown with reference to Figures 23 to 36. In this embodiment, the first actuator 150 is configured to move from a first unpressed position to a second pressed position to aspirate fluid into the pipette tip to which the fluid is attached (see Figures 26 and 33). In addition, the second actuator 155 is configured to move from the first pressed position to the second unpressed position when the first actuator 150 moves from the first unpressed position to the second pressed position, so that the fluid contained in the pipette tip can be electroporated by moving the second actuator 155 from the second unpressed position to the first pressed position and then dispensed. By moving the first actuator 150 from a first non-pressed position to a second pressed position, an aspiration function (for example, aspiration of fluid into the pipette tip attached to the distal section of the pipette) is generated, and by moving the second actuator 155 from a second non-pressed position to a first pressed position, a dispensing function (for example, dispensing fluid from the pipette tip attached to the distal section of the pipette) is generated.

[0059] In various embodiments, the pipette includes a lock button 165 disposed on the proximal section 135 of the pipette, as shown, for example, in Figures 2, 9, 23, and 31. The lock button 165 is operably connected to a locking mechanism configured to control the locking of a plurality of gripper mechanisms, as will be discussed in more detail herein. When the lock button is pressed, the locking mechanism transitions from a first unlocked configuration to a second locked configuration. When the locking mechanism is in the first configuration, the plurality of gripper mechanisms are open and configured to receive the engaging sections of the plungers, and when the locking mechanism is in the second configuration, the plurality of gripper mechanisms are closed and configured to grip and engage with the engaging sections of the plungers at the tip of the pipette. In some embodiments, the locking mechanism transitions from the second locked configuration to the first unlocked configuration when a third actuator 160 is activated (e.g., pressed).

[0060] Figures 23–30 show embodiments of the pipette 130 including a gear mechanism 170 having a specific configuration of operably connected gears. Figures 31–36 show another embodiment of the pipette 130 including a gear mechanism 170 having a different configuration of gears compared to the gear mechanism 170 shown in Figures 23–30. However, both gear mechanism configurations provide similar aspiration and dispensing functionality, achieved by the same manual operation of the first actuator 150, second actuator 155, third actuator 160, and lock button. In addition, Figure 66 shows another embodiment of the gear mechanism 170 shown for the pipette 130 in Figures 31–36, where the gears are fixed to a single shaft to allow frictionless gear rotation.

[0061] Figures 25 and 32 illustrate the movement of individual gears in the transition of the second actuator 155 for dispensing fluid to the first depressed position. Figures 26 and 33 illustrate the movement of individual gears in the transition of the first actuator 150 for aspirating fluid to the second depressed position.

[0062] Additional components of the pipette 130 are shown through Figures 23 to 36. In various embodiments, the pipette 130 includes a shaft 175, a linkage rack 174, a push plate 176, a linkage bar 178, a bush 180, and a plurality of gripper mechanisms 182, each gripper mechanism having a gripper jaw 184 movably disposed within a gripping sleeve 186. The linkage bar 178 is rigidly connected to the push plate 176, which operates to contact the individual gripper jaws 184 and the proximal end of each gripping sleeve 186. The linkage rack 174 is operably connected to the push plate 176 and transmits the movement of the first and second actuators (150, 155) and the pressing of the lock button 165 to the linkage bar 178 during the performance of aspiration, dispensing, and locking functions. The bushing 180 is operably connected to the linkage bar 178 and engages with the shaft 172 so as to have minimal resistance during the linear movement of the linkage bar 178 along the length of the shaft during operation. This allows for highly precise and flexible movement.

[0063] Referring to Figures 42–44, in some embodiments, the pipette 130 also includes magnets configured to interact with each other to enable certain functionalities. In various embodiments, the pipette 130 includes an outer housing 900 which includes a separate magnet 910 that interacts with a separate magnet 910 disposed on the internal components of the pipette, and the magnetic interaction achieves a specific function. Figures 42–44 illustrate portions of the pipettes depicted in Figures 2–15 and Figures 31–36. In such embodiments, the outer housing 900 includes magnets 910 disposed within or on the outer housing to interact with magnets disposed within or on the linkage bar 178 of the pipette. Figure 42 shows the location of magnets disposed on the outer housing of the pipette that magnetically interact with magnets disposed on the linkage bar 178, as shown in Figure 43. As shown in Figure 44, after the pipette has performed its aspiration function (for example, by moving the first actuator from a first non-pressed position to a second pressed position), the linkage bar 178 is magnetically held in a position such that the push plate 176, the bush 180, and the components of the gripper mechanism (e.g., the gripper jaws 184 and the gripping sleeve 186) are oriented toward the proximal section of the pipette along the length of the shaft. In this configuration, a large portion of the length of the individual plungers of the mounted pipette tip held within the gripper jaws 184 is also withdrawn from the lumen of the pipette tip containing the aspirated fluid. The pipette 130 is held in this configuration while the pipette is inserted into the pipette docking assembly 110 and the electroporation procedure is performed.

[0064] Referring to Figures 25 and 32, the pipette described herein includes a plurality of gripper mechanisms 182 disposed in the distal section 145 of the pipette. In embodiments, each gripper mechanism 182 is configured to engage with a component of a pipette tip 200 suitable for performing electroporation. The pipette tip is first attached to the distal section of the pipette via a clip-on connection formed by the engagement of a tab on the pipette tip mounting interface with a mounting feature surrounding a portion of the gripper mechanism. The plunger of the pipette tip is then grasped and locked by a gripper mechanism that allows control of the linear motion of the plunger within the lumen of the pipette tip via the operation of first and second actuators and a lock button on the pipette in order to perform the pipette function.

[0065] As discussed herein, each pipette tip for use with the pipettes of this disclosure includes a clip-on connection with a plunger disposed within the lumen of the pipette tip, which is made of a conductive material to facilitate the flow of current through the sample contained in the lumen of the pipette tip during use for carrying out cell transfection protocols.

[0066] As shown in Figures 30 and 36, each gripper mechanism is configured to reversibly grip the plunger 402 of the pipette tip 200 via the engagement section of the plunger when the pipette tip is attached to the gripper mechanism via a clip-on connection. When the engagement section of the plunger is gripped by the gripper mechanism and the locking mechanism is activated (for example, in the second locking configuration via pressing a lock button), the movement of the plunger 402 in the lumen of the pipette tip is controlled by the first and second actuators to perform pipetting functions, such as aspirating and dispensing fluid into and out of the lumen of the pipette tip.

[0067] Figures 37 to 41 illustrate the components of a gripper mechanism in several embodiments. Each gripper mechanism 182 includes a gripper jaw 184 having a jaw opening 185 for receiving and holding the engagement section 404 of a plunger, and a gripping sleeve 186 positioned around the gripper jaw. The gripping sleeve 186 is configured to exert an inward force on the gripper jaw 184 such that when a lock button is pressed, it causes the gripper jaw to exert an inward force on the engagement section of the plunger in order to hold the engagement section of the plunger within the gripper jaw.

[0068] Figures 38 and 39 show the gripper mechanism 182 in an open configuration suitable for receiving the engagement section 404 of the plunger. Figures 40 and 41 show the gripper mechanism 182 in a closed configuration suitable for gripping the engagement section 404 of the plunger. In particular, distal movement of the gripping sleeve 186 relative to the gripper jaws 184 moves the gripper mechanism 182 from the open configuration to the closed configuration. This movement is achieved by pressing the lock button.

[0069] Figure 36 is a cross-sectional view of a gripper mechanism for gripping the engagement section of a pipette tip plunger attached to the tip interface via the pipette tip mounting interface. Surrounding the gripping sleeve 186 is the tip interface 188, which is configured to engage with the pipette tip mounting interface 806 (see Figure 22) and includes a retaining platform 955 that engages with the tab 808 (see Figure 22) of the mounting interface to facilitate clip-on connection. In embodiments, the retaining platform 955 is distally positioned on the tip interface and encircles the outer surface of the tip interface 188, as further discussed herein.

[0070] As shown in Figure 36, the tip interface 188 includes mounting features, such as a retaining platform 955, for engaging with the tab 808 of the mounting interface 806. Figure 36 shows the retaining platform 955 positioned on the outer surface of the tip interface 188 in the distal section of the pipette. The retaining platform 955 rests on the inclined surface 960 of the tip interface and is defined by a flat annular surface that traverses the circumference of the tip interface 188, the surface extending radially at an angle of 90 degrees from the longitudinal axis of the tip interface such that the annular surface of the retaining platform is perpendicular to the longitudinal axis of the tip interface and the longitudinal axis of the pipette. During pipette tip mounting, as the distal section 145 of the pipette 130 is pushed into the pipette tip mounting interface 806, the tab 808 of the mounting interface may advance on the inclined surface 960 and extend until it reaches the retaining platform 955, at which point the tab 808 may retract inward toward the longitudinal axis of the tip interface and engage with the retaining platform 955.

[0071] In some examples, after the tab 808 reaches the retaining platform 955 and retracts toward the central axis of the tip interface 188, the biasing member of the pipette 130 may act to bias the tab 808 to engage with the retaining platform 955. Figure 35 shows the pipette tip 200 attached to the tip interface and the biasing member 962 of the distal section 145 of the pipette 130, including the biasing platform 964. A spring 966 is operably connected to the biasing member 962 and functions to align the pipette tip perpendicular to the biasing platform 964.

[0072] Figure 36 shows the pipette tip mounting interface 806 engaging with the holding platform 955 of the gripping sleeve 186. During pipette tip mounting, as the mounting interface 806 advances over the tip interface 188, the mounting interface 806 pushes and moves the biasing platform 964, allowing the tab 808 to reach the holding platform 955. After the mounting interface 806 stops pushing the pipette 130 into the distal section 145 (after the tab 808 has reached the holding platform), the spring 966 compresses responsively, pressing the biasing platform 964 against the mounting interface 806, engaging its tab 808 with the holding platform, and aligning the pipette tip perpendicular to the biasing platform.

[0073] Figure 36 shows the engagement section 404 of the plunger at the tip of the pipette that engages with the gripper jaw 184 of the gripper mechanism. During pipette operation, the engagement section 404 is locked and gripped by the gripper mechanism in the distal section 145 of the pipette 130 by activating (e.g., pressing) a lock button located in the proximal section 135 of the pipette 130. In this embodiment, a second actuator is operable to advance the gripper jaw 184 distally and engage with the engagement section 404 of the plunger at the tip of the pipette, which is attached to the pipette via the tip interface 188. During pipette operation, the gripper mechanism is activated by pressing the lock button to lock the engagement section of the plunger in the gripper jaw. The movement of the plunger is then controlled by the operation of the first and second actuators to perform aspiration and dispensing functions.

[0074] In one embodiment, the gripper jaw advances to engage with the engagement section 404 simultaneously with the tab 808 of the pipette tip mounting interface 806 advancing to engage with the retaining platform 955 (for example, the locking mechanism is in a first configuration (via the operation of a third actuator) while the distal section of the pipette and the pipette tip mounting interface are pushed against each other by having a second actuator in a first depressed position). Alternatively, the gripper jaw advances to engage with the engagement section 404 asynchronously with the tab advancing to engage with the retaining platform 955 by first pushing the tab 808 to engage with the retaining platform 955, and then moving the second actuator to a first depressed position. Alternatively, the gripper jaw can be advanced to engage with the engaging section 404 asynchronously with the tab 808 being advanced to engage with the holding platform 955 by moving the second actuator to the first depressed position, followed by moving the first actuator to the first depressed position, and then pushing the tab 808 to engage with the holding platform 955.

[0075] The gripper mechanism includes a gripping sleeve 186 positioned around the gripper jaw 184. The gripping sleeve 186 is configured to exert an inward force on the gripper jaw 184, causing the gripper jaw 184 to exert an inward force on the engagement section 404, thereby holding the engagement section of the plunger within the gripper jaw 184.

[0076] After gripping and locking the plunger, which engages with the gripper mechanism (by pressing the lock button), the plunger is controlled by first and second actuators to perform the pipetting function. Once the plunger is locked by the gripper jaws, the sample may be drawn into the pipette tip or dispensed from the pipette tip.

[0077] The operation of the pipette for electroporating a sample is described in embodiments of this disclosure as follows: First, the technician grasps the pipette handle and holds the pipette with one hand. Next, the second actuator is moved to the first pressed position, moving the gripper jaws of each gripper mechanism fully distally and positioning them to receive the engagement section of the pipette tip (see Figures 25 and 32). This action causes the magnet located on the outer housing of the pipette to magnetically align with the magnet located on the linkage bar, holding the first actuator in the first unpressed position (see Figure 52). Next, the third actuator is actuated by pressing the actuator, unlocking the gripper mechanism, thereby moving the locking mechanism operably connected to the third actuator to the second unlocked configuration. In particular, the gripper mechanism remains in the first unlocked and open configuration when the third actuator is pressed without additional manual operation, as a result of the unique design of the gripper mechanism which allows it to be held in the open configuration without requiring the technician to continuously hold the button in the pressed position.

[0078] The technician then moves the pipette over the pipette tip, aligning the distal section of the pipette with the pipette tip and pushing the gripper jaws and surrounding tip interface into the mounting interface of the individual pipette tip. This action forms a clip-on connection so that the pipette tip is attached to the distal section of the pipette and the engaging section of the plunger at the pipette tip is received by the open gripper jaws. The locking mechanism is then switched to a second locking configuration by pressing the lock button, in which the gripper mechanism grips the plunger at the pipette tip, locking the plunger in a gripping engagement with the pipette.

[0079] Next, while the distal tip of the pipette tip is in contact with the liquid sample, the liquid sample is drawn into the pipette tip by moving the first actuator from a first unpressed position to a second pressed position. The pipette is then fixed to the pipette docking assembly of the electroporation system described herein so that the liquid sample contained within the lumen of the pipette tip is in contact with the electrolytic buffer contained within the reservoir of the docking assembly. The liquid sample is electroporated by a current generated by a pulse generator, which is electrically connected to the electrolytic buffer in the reservoir via a first electrode electrically connected to the pulse generator, and by delivering a conductive plunger at each pipette tip to the sample via a second electrode contained within the pipette and electrically connected to the pulse generator. The electroporated sample is then dispensed from the pipette tip into a desired container by moving the second actuator from a second unpressed position to a first pressed position.

[0080] After the sample has been dispensed, the pipette tip can be disengaged (e.g., ejected) from the pipette by a one-step or two-step process. In a one-step process, the pipette tip is disengaged by activating a third actuator by fully depressing the actuator. Alternatively, a two-step process may be used to reduce the amount of peak force required to disengage the pipette tip, which involves first partially depressing the third actuator to separate the tabs of each retaining interface of the attached pipette tip from the pipette's retaining platform while the plunger remains grippingly locked in the gripper jaws, and then fully depressing the third actuator to transition the locking mechanism from a second locking configuration to a first unlocking configuration (gripper jaws open) so that the engaging section of the plunger is released from the gripper jaws. In embodiments, a one-step ejection process may require a peak force of about 60 N, and a two-step ejection process such as the one described herein may require a peak force of about 40 N.

[0081] As discussed herein, the pipette 130 includes a third actuator that is operable to facilitate the disengagement of the pipette tip attached to the distal section of the pipette. As shown throughout the figures, the third actuator 160 is located in the proximal section 135 of the pipette 130 and, when the third actuator 160 is actuated, is configured to disengage the pipette tip attached to the distal section 145 of the pipette 130 (for example, by being fully pressed for disengagement of the pipette tip via a one-step process, or by being partially pressed and then fully pressed for disengagement of the pipette tip via a two-step process).

[0082] In both processes, partial depression of the third actuator causes the tip ejection sleeves, positioned around each tip interface in the distal section of the pipette, to move distally relative to the tip interface, displacing the pipette tip tabs engaged with the tip interface retaining platform, thereby disengaging the pipette tip mounting interface from the distal section of the pipette (without disengaging the gripper mechanism from the plunger). Simultaneously with the disengagement of the pipette tip mounting interface, the biasing platform in contact with the mounting interface is pushed distally via a force exerted by a compression spring operably connected to the biasing platform, ejecting the mounting interface distally.

[0083] Furthermore, in both processes, fully pressing the third actuator also transitions the locking mechanism from the second locked configuration to the first unlocked configuration, thereby transitioning the gripper mechanism to the open configuration, disengaging the engaging section of the plunger at the pipette tip and releasing it from the gripper jaws. It will be understood that after the third actuator is fully pressed, the entire pipette tip (including, for example, the plunger and mounting interface) is disengaged from the pipette.

[0084] Figures 45–49 illustrate the disengagement of the pipette tip from the pipette embodiment shown in Figure 31 (e.g., ejection of the pipette tip) using a one-step process. With the second actuator 155 in the first depressed position (e.g., after the dispensing function has been performed), fully pressing the third actuator 160 (e.g., the ejection button) causes the tip ejection sleeve 190 (see Figure 49) to move distally relative to the tip interface 188, disengaging the mounting interface via a distal force transmitted through the clip tip release plate 750, and causing disengagement of the plunger engagement section by proximal movement of the gripping sleeve relative to the distal end of the gripper jaw, which is caused by distal movement of the plunger release plate 760 resulting in proximal movement of the lifter plate 770, which occurs only when the third actuator is fully pressed. The separation of the pipette tip, which is completely disengaged from the distal section of the pipette, is facilitated by the distal force exerted on the pipette tip via the biasing platform.

[0085] Figures 51–53 illustrate the disengagement of the pipette tip from the pipette embodiment shown in Figure 31 (e.g., pipette tip ejection) using a two-step process. With the second actuator in the first depressed position (e.g., after the dispensing function has been performed), the third actuator (e.g., the ejection button) is partially pressed, causing the tip ejection sleeve (see Figure 49) to move distally relative to the tip interface and disengage the attachment interface via a distal force transmitted through the release plate. Figure 53 illustrates the partial disengagement of the pipette tip after the third actuator has been partially pressed and the tip ejection sleeve has detached the pipette tip attachment interface from the distal section of the pipette. The separation of the partially disengaged pipette tip attachment interface from the distal section of the pipette is facilitated by a distal force exerted on the pipette tip via the biasing platform. Next, the third actuator is fully depressed to disengage the plunger engagement section by the proximal movement of the gripping sleeve relative to the distal end of the gripper jaw, which is caused by the distal movement of the plunger release plate, resulting in the proximal movement of the lifter plate that occurs only when the third actuator is fully depressed.

[0086] As discussed herein, when the third actuator 160 is partially or completely pressed, it can cause the tip discharge sleeve 190 to actuate, thereby advancing the tip discharge sleeve distally (e.g., downward) from the inside of the tab 808 toward the tab 808. During distal advancement, the discharge sleeve 190 may push the inside of the tab 808, bending the tab 808 outward and disengaging the tab 808 from the retaining platform 955. The biasing member 962 then pushes the mounting interface 806 of the pipette tip 802 downward from the pipette 130 (via contact with the biasing platform 964), thereby discharging the mounting interface 806 of the pipette tip 802 (and the lumen attached thereto) from the pipette.

[0087] In this embodiment, the third actuator 160 is configured to traverse the blank travel distance when pressed, before causing disengagement of the mounting interface 806 from the pipette 130 and the lumen of the pipette tip (this occurs by pressing the third actuator 160 to its final distance) (see Figures 46 and 58). Such functionality is designed to prevent accidental discharge of the mounting interface and lumen of the pipette tip from the pipette 130.

[0088] Pipette docking assembly

[0089] In many existing electroporation systems, the reservoir holding the electrolytic buffer (also referred to herein as the “buffer tube”) can be easily detached from the pipette station (also referred herein as the “pipette docking station”, “pipette station”, or “docking station”), so that the reservoir may be inadvertently detached from the docking station when withdrawing the pipette tip from the reservoir. Such inadvertent detachment may lead to spillage and / or damage to the pipette tip. In some examples, the reservoir (e.g., the buffer tube) is held in place by a pipette station guard (also referred to herein as the “station guard”) associated with the docking station (e.g., to protect the user from electric shock), but conventional pipette station guards may also be inadvertently detached during pipette withdrawal from the reservoir (or even during electroporation, which may present a risk of electric shock).

[0090] In embodiments, the disclosure provides a pipette station guard that locks to a docking station via movement of the station guard in a locking direction different from the pipette removal direction for removing a pipette from a reservoir. The reservoir is inserted into an opening in the station guard and locked to the station guard. The reservoir can be released from the station guard by applying force in the same or different force application direction as the pipette removal direction for removing a pipette from the reservoir (e.g., to a latching member). Such features can reduce or eliminate the occurrence of accidental removal of the reservoir (or other reservoir) and / or the station guard from the pipette docking station during pipette removal, thereby reducing or avoiding spills and / or damage to the pipette tip.

[0091] Figure 59 illustrates an electroporation system 10 of the present disclosure, which includes a pipette docking assembly 110 electrically coupled to a pulse generator 100. Figure 59 also shows a pipette 130 coupled to the pipette docking assembly 110, positioned to perform an electroporation procedure. The distal section (and attached pipette tip) of the pipette 130 is docked within the pipette docking assembly 110 so that the pipette is received into the reservoir 125 and subsequently by the station guard 120.

[0092] Figures 60 and 61 illustrate the front and rear views of the pipette docking assembly 110 shown in Figure 59, with the pipette 130 and reservoir 125 not docked.

[0093] Figure 62 illustrates an embodiment of a reservoir 125 used with the pipette docking assembly 110 shown in Figure 59. In this embodiment, the reservoir 125 includes a plurality of electrodes 117, typically one electrode corresponding to each channel of the pipette. The reservoir shown in Figure 62 includes eight electrodes 117 for use with a pipette configured to use eight pipette tips. It will be understood that the reservoir 125 may include 2 to 16 electrodes for use with a pipette configured to use a corresponding number of pipette tips. The electrodes 117 of the reservoir are electrically connected to the pulse generator 100 via electrodes disposed on the docking station 115 when the reservoir 125 is docked to the docking station 115 together with the station guard 120.

[0094] Figure 63 illustrates a pipette station guard 120 in an embodiment of the present disclosure. The pipette station guard 120 may be attached to a pipette docking station 115 to protect the user from potential electric shock. The pipette station guard 120 may also include a reservoir opening for receiving a buffer reservoir (e.g., reservoir 125) that can receive a pipette (e.g., pipette 130) and / or components connected thereto (e.g., consumable pipette tip).

[0095] To facilitate connection to the pipette docking station 115, the pipette station guard 120 may include various connecting elements, such as one or more locking hooks configured to engage with one or more corresponding hook catches of the pipette docking station. Such locking hooks may take various forms. For example, the embodiment in Figure 63 illustrates the pipette station guard 120 as including one or more pivot hooks 122 configured to rotate to engage with one or more hook catches (e.g., pivot hook catches) of the pipette docking station (see Figure 64).

[0096] In the embodiment shown in Figure 63, the pipette station guard 120 includes a pair of pivot hooks 122 located on the rear surface of the pipette station guard 120. The pair of pivot hooks 122 are located on the upper portion of the rear surface.

[0097] Figure 64 illustrates a pipette station guard 120 being moved to engage with a docking station 115. After the pivot hook 122 is inserted into the receiving point of the docking station, the station guard 120 rotates until the opposing latch 124 reaches the corresponding catch and engages, as shown in Figure 65.

[0098] Although the present invention has been described in considerable detail with reference to certain preferred embodiments, other versions are also possible. Therefore, the spirit and scope of the appended claims should not be limited to the descriptions and preferred versions contained herein. Various aspects of the present invention are illustrated with reference to the following non-limiting embodiments.

[0099] Any modifications and / or alterations to the features of the invention illustrated herein, as well as additional uses of the principles illustrated herein, made by persons skilled in the relevant art and owners of this disclosure, may be made to the illustrated embodiments without departing from the spirit and scope of the invention as defined by the claims, and should be considered within the scope of this disclosure. Accordingly, although various aspects and embodiments are disclosed herein, other aspects and embodiments are contemplated. Numerous methods and components similar or equivalent to those described herein may be used in particular embodiments of this disclosure, but only certain specific components and methods are described herein.

[0100] Furthermore, it will be understood that a system, process, and / or product according to a particular embodiment of the present disclosure may include, incorporate, or otherwise incorporate properties (e.g., components, members, elements, parts, and / or parts) described in other embodiments disclosed and / or described herein. Accordingly, various features of a particular embodiment may be compatible with, combined with, included in, and / or incorporated in other embodiments of the present disclosure. Therefore, the disclosure of a particular feature relating to a particular embodiment of the present disclosure should not be interpreted as limiting the application or inclusion of that feature in that particular embodiment. Rather, it will be understood that other embodiments may include that feature without necessarily departing from the scope of the present disclosure.

[0101] Furthermore, unless a feature is described as requiring another feature in combination with it, any feature herein can be combined with any other feature of the same or different embodiments disclosed herein. Moreover, various well-known embodiments of exemplary systems, processes, products, etc., are not described in particular detail herein to avoid obscuring the embodiments of the exemplary models. However, such embodiments are contemplated herein as well.

[0102] This disclosure may be embodied in other specific forms without departing from the spirit or essential features of the invention. The embodiments described are, in all respects, illustrative only and not restrictive. Accordingly, the scope of the invention is indicated by the appended claims rather than by the foregoing description. While certain embodiments and details are included herein and in the appended disclosure for the purpose of illustrating embodiments of this disclosure, it will be apparent to those skilled in the art that various modifications are possible in the methods, products, devices, and apparatus disclosed herein without departing from the scope of this disclosure or the invention as defined in the appended claims. All modifications that fall within the meaning and scope of equivalent claims are encompassed within that scope.

[0103] The exemplary subject matter of the present invention is represented by the following clauses.

[0104] Clause 1: A multichannel pipette, A proximal section having a handle, A distal section configured to reversibly engage with multiple pipette tips, A first actuator is located in the proximal section, It comprises a second actuator located in the proximal section and capable of operating to control the dispensing function of the multichannel pipette, A multi-channel pipette in which a first actuator is operable to control aspiration function and a second actuator is operable to control dispensing function.

[0105] Clause 2: The multichannel pipette of Clause 1, further comprising a third actuator disposed in the proximal section, the third actuator being configured to disengage a pipette tip attached to the distal section of the multichannel pipette when the third actuator is actuated.

[0106] Clause 3: A multichannel pipette according to Clause 2, wherein a first actuator is configured to move from a first non-pressed position to a second pressed position, and a second actuator is configured to move from a first pressed position to a second non-pressed position when the first actuator moves from a first non-pressed position to the second pressed position, thereby producing an aspiration function by moving the first actuator from a first non-pressed position to a second pressed position, and a dispensing function by moving the second actuator from a second non-pressed position to a first pressed position.

[0107] Clause 4: A multichannel pipette according to Clause 3, wherein the aspiration function includes the aspiration of fluid into the pipette tip engaged with the distal section of the multichannel pipette.

[0108] Clause 5: A multichannel pipette according to Clause 3, wherein the dispensing function includes dispensing fluid from a pipette tip engaged with the distal section of the multichannel pipette.

[0109] Clause 6: The multichannel pipette of Clause 2, further comprising a plurality of gripper mechanisms disposed in the distal section, each gripper mechanism configured to reversibly grip a plunger disposed in the lumen of the pipette tip, wherein the plunger has an engagement section for engaging with a gripper mechanism and a lumen section disposed in the lumen of the pipette tip.

[0110] Clause 7: A multichannel pipette according to Clause 6, further comprising a lock button disposed in the proximal section, operably connected to a locking mechanism configured to control the locking of a plurality of gripper mechanisms, wherein pressing the lock button causes the locking mechanism to transition from a first unlocked configuration to a second locked configuration, wherein when the locking mechanism is in the first configuration, the plurality of gripper mechanisms are configured to open and receive the engaging sections of plungers, and when the locking mechanism is in the second configuration, the plurality of gripper mechanisms are configured to close and grip and engage with the engaging sections of plungers.

[0111] Clause 8: The locking mechanism of the multichannel pipette of Clause 7 transitions from a second locked configuration to a first unlocked configuration when a third actuator is activated.

[0112] Clause 9: Each of the multiple gripper mechanisms, A gripper jaw comprising a jaw opening for receiving and holding an engaging section, A multichannel pipette according to Clause 7 or Clause 8, comprising a gripping sleeve positioned around a gripping jaw, wherein the gripping sleeve is configured to exert an inward force on the gripping jaw such that the gripping jaw exerts an inward force on the engaging section of the plunger in order to hold the engaging section of the plunger within the gripping jaw when the locking mechanism is in a second locking configuration.

[0113] Clause 10: The multichannel pipette according to Clause 9, wherein pressing the lock button moves the gripping sleeve distal to the gripper jaws, thereby applying an inward force to the gripper jaws.

[0114] Clause 11: The multichannel pipette of Clause 10, wherein by activating a third actuator, the gripping sleeve moves proximal to the gripper jaws, thereby retracting the inward force on the gripper jaws.

[0115] Clause 12: The multichannel pipette according to Clause 11, wherein the first actuator comprises a button located at the proximal end of an elongated shaft, and the second actuator comprises a button located at the proximal end of an elongated shaft.

[0116] Clause 13: A multichannel pipette according to Clause 12, wherein a lock button is located at the distal end of the shaft, the shaft of the lock button is parallel to the shaft of the second actuator, and when the second actuator moves from a first pressed position to a second unpressed position, or from a second unpressed position to a first pressed position, the lock button and the shaft of the lock button translate to the button of the second actuator and the shaft of the second actuator.

[0117] Clause 14: A multichannel pipette according to Clause 13, comprising a locking mechanism with a push plate that reversibly contacts the proximal end of the gripping sleeve, wherein pressing the lock button causes the push plate to move the gripping sleeve distally relative to the gripper jaws.

[0118] Clause 15: The multichannel pipette of Clause 14, wherein by activating a third actuator, the tip discharge sleeve moves distally to the gripper jaws and contacts the proximal end of the pipette tip, which is circumferentially arranged around the gripper jaws, thereby disengaging the pipette tip.

[0119] Clause 16: A multichannel pipette according to Clause 15, wherein the locking mechanism is in a second configuration, and while engaged with the engaging section of the plunger, the plunger moves proximal to the gripper jaw by moving the first actuator from a first non-pressed position to a second pressed position, thereby drawing fluid into the pipette tip via the suction force generated by the proximal movement of the lumen section within the lumen of the pipette tip.

[0120] Clause 17: A multichannel pipette according to Clause 16, wherein the locking mechanism is in a second configuration and, while engaged with the engaging section of the plunger, moves a second actuator from a second non-pressed position to a first pressed position, thereby moving the plunger distally relative to the gripper jaw, thereby dispensing fluid from the pipette tip via a displacement force generated by the distal movement of the lumen section within the lumen of the pipette tip.

[0121] Clause 18: A multichannel pipette according to Clause 6, having two, three, four, five, six, seven, or eight or more gripper mechanisms.

[0122] Clause 19: The multichannel pipette according to Clause 18, which has eight gripper mechanisms.

[0123] Clause 20: A multichannel pipette according to any one of the preceding clauses, further comprising a tip interface arranged circumferentially around each gripper jaw, the tip interface being configured to engage with tabs of a pipette tip retaining interface to secure the pipette tip to the pipette.

[0124] Clause 21: A multichannel pipette according to any one of the preceding clauses, further comprising a tip discharging sleeve disposed on the tip interface, which, when a third actuator is activated, moves distally to the tip interface and is operable to displace the pipette tip attachment interface, thereby detaching the pipette tip from the pipette.

[0125] Clause 22: A multichannel pipette according to any one of the preceding clauses, further comprising an electrode disposed in the distal section, wherein the electrode is electrically coupled to each gripper jaw of a plurality of gripper mechanisms.

[0126] Clause 23: A multichannel pipette according to Clause 22, wherein each gripper jaw is made of a conductive material and is operable to allow an electrical pulse applied to an electrode to pass through the electrode, through each gripper jaw, through the plunger at the pipette tip, through the sample contained in the lumen of the pipette tip, and through a second electrode positioned adjacent to the distal end of the pipette tip, thereby enabling electroporation of cells contained in the sample.

[0127] Clause 24: An electroporation system, A multichannel pipette according to any one of clauses 1 to 23, The tip of the pipette and Pipette docking assembly and An electroporation system equipped with a pulse generator.

[0128] Clause 25: The electroporation system of Clause 24, wherein the pipette docking assembly comprises a pipette station, a pipette station guard, and a reservoir.

[0129] Clause 26: The electroporation system of Clause 25, including the reservoir and buffer.

[0130] Clause 27: An electroporation system according to any one of Clauses 24 to 26, wherein the pipette tip is equipped with a 10 μL pipette tip or a 100 μL pipette tip.

[0131] Clause 28: An electroporation system according to any one of Clauses 24-27, wherein the pipette tip comprises a plunger at least partially disposed within the lumen, and the plunger is configured to translate along the lumen to facilitate aspiration and / or dispensing.

[0132] Clause 29: An electroporation system according to Clause 28, wherein the plunger comprises gold, diamond-like carbon, and / or conductive plastic.

[0133] Clause 30: An electroporation system according to Clause 28 or Clause 29, wherein the plunger comprises an engaging section and a lumen section, the lumen section comprising a sealing component for creating a seal between the plunger and the lumen.

[0134] Clause 31: The electroporation system of Clause 30, wherein the lumen section comprises a front pin and a shaft section, the front pin being configured to connect to the shaft section and to secure a sealing component to the shaft section.

[0135] Clause 32: The electroporation system of Clause 31, wherein the sealing component comprises a polymer sleeve, and a front pin is configured to be inserted through the polymer sleeve and engage with a retaining hole in the shaft section to secure the sealing component to the shaft section.

[0136] Clause 33: The electroporation system of Clause 32, wherein the insertion of the front pin through the polymer sleeve defines a space between the polymer sleeve and the front pin, and this space contributes to the flexibility of the polymer sleeve to create a seal between the plunger and the lumen.

[0137] Clause 34: An electroporation system according to any one of Clauses 30 to 33, wherein the sealing component includes polytetrafluoroethylene (PTFE).

[0138] Clause 35: The electroporation system of Clause 30, wherein the sealing component comprises a coated O-ring, and the lumen section includes a circumferential recess configured to receive the coated O-ring.

[0139] Clause 36: An electroporation system according to any one of Clauses 30 to 35, wherein the pipette tip further comprises a mounting interface adjacent to the lumen, and the mounting interface comprises one or more tabs configured to engage with a retaining platform of the distal section of the pipette assembly.

[0140] Clause 37: An electroporation system according to any one of Clauses 24 to 36, The pipette tip is connected to a multichannel pipette and positioned in contact with the buffer within the reservoir. The first electrode is electrically coupled to the buffer. The plunger at the tip of the pipette is electrically connected to the sample inside the pipette tip and the pipette electrode. An electroporation system according to any one of clauses 24 to 36, wherein the pipette electrode is in contact with the second electrode, and the first electrode and the second electrode are electrically connected to a pulse generator.

[0141] Clause 38: An electroporation system according to any one of Clauses 24 to 37, comprising one or more voltage sources configured to charge one or more high-voltage capacitors, the one or more high-voltage capacitors configured to operate as a power source for an amplifier circuit, the amplifier circuit configured to supply voltage to a sample in a multichannel pipette in a manner that takes into account load variations.

[0142] Clause 39: The electroporation system of Clause 38, wherein the amplifier circuit comprises a common source amplifier configured to output high-voltage pulses.

[0143] Clause 40: An electroporation system according to Clause 39, in which a common source amplifier receives a signal from an amplitude setting loop, the signal from the amplitude setting loop is based on inputs from a digital-to-analog converter and an input from a voltage sensing loop, the input from the voltage sensing loop is determined using high-voltage pulses, a voltage divider, and a differential amplifier.

[0144] Clause 41: The electroporation system of Clause 40, wherein a common source amplifier amplifies the signal from the amplitude setting loop by a coefficient of approximately 1,000 to approximately 2,000.

[0145] Clause 42: An electroporation system according to Clause 40 or Clause 41, wherein the input from the digital-to-analog converter corresponds to a waveform selected by the user.

[0146] Clause 43: An electroporation system according to any one of Clauses 24 to 42, comprising an arc discharge detection module configured to detect an arc discharge within a sample while a pulse generator applies voltage to the sample.

[0147] Clause 44: The arc discharge detection module, A first step amplifier configured to provide an amplified current signal based on the current signal associated with the voltage applied to the sample, A bandpass filter configured to filter a falling edge signal from an amplified current signal, wherein the falling edge signal indicates a decrease in current passing through the sample, and the decrease in current suggests an arc discharge. An electroporation system according to Clause 43, comprising: a comparator configured to determine whether an arc discharge occurred in a sample by comparing a falling edge signal filtered by a bandpass filter with one or more reference standards.

[0148] Clause 45: The electroporation system of Clause 44, wherein the amplification of the current signal by a first-stage amplifier is based on the output of a low-voltage detection circuit for determining the resistance associated with the sample.

[0149] Clause 46: A method for transfecting cells with a payload, To provide an electroporation system as described in any one of clauses 24 to 45, Providing cells and To provide a payload, Introducing cells and payload into the tip of the pipette, A method comprising electroporating cells by operating an electroporation system.

[0150] Clause 47: The method of Clause 46, wherein the cells are mammalian cells.

[0151] Clause 48: The method of Clause 46, wherein the cells are microorganisms or organoids.

[0152] Clause 49: The method of Clause 46, wherein the payload is selected from the group consisting of nucleic acids, proteins, or combinations thereof.

Claims

1. It is a multichannel pipette, A proximal section having a handle, A distal section configured to reversibly engage with multiple pipette tips, A first actuator disposed in the aforementioned proximal section, The pipette comprises a second actuator disposed in the proximal section and capable of operating to control the dispensing function of the multichannel pipette, A multichannel pipette in which the first actuator is operable to control a suction function and the second actuator is operable to control the dispensing function.

2. The multichannel pipette according to claim 1, further comprising a third actuator disposed in the proximal section, wherein the third actuator is configured to disengage the pipette tip attached to the distal section of the multichannel pipette when the third actuator is activated.

3. The multichannel pipette according to claim 2, wherein the first actuator is configured to move from a first non-pressed position to a second pressed position, and the second actuator is configured to move from a first pressed position to a second non-pressed position when the first actuator moves from a first non-pressed position to a second pressed position, and a suction function is generated by moving the first actuator from a first non-pressed position to a second pressed position, and a dispensing function is generated by moving the second actuator from a second non-pressed position to a first pressed position.

4. The multichannel pipette according to claim 3, wherein the aspiration function includes aspirating fluid into the pipette tip engaged with the distal section of the multichannel pipette.

5. The multichannel pipette according to claim 3, wherein the dispensing function includes dispensing a fluid from a pipette tip engaged with the distal section of the multichannel pipette.

6. The multichannel pipette according to claim 2, further comprising a plurality of gripper mechanisms disposed in the distal section, each gripper mechanism configured to reversibly grip a plunger disposed in the lumen of the pipette tip, wherein the plunger has an engagement section for engaging with the gripper mechanism and a lumen section disposed in the lumen of the pipette tip.

7. The multichannel pipette according to claim 6, further comprising a lock button disposed in the proximal section, which is operably connected to a locking mechanism configured to control the locking of the plurality of gripper mechanisms, wherein when the lock button is pressed, the locking mechanism transitions from a first unlocking configuration to a second locking configuration, and when the locking mechanism is in the first configuration, the plurality of gripper mechanisms are configured to open and receive the engaging sections of the plungers, and when the locking mechanism is in the second configuration, the plurality of gripper mechanisms are configured to close and grip and engage with the engaging sections of the plungers.

8. The multichannel pipette according to claim 7, wherein the locking mechanism transitions from the second lock configuration to the first unlock configuration when the third actuator is activated.

9. Each of the aforementioned plurality of gripper mechanisms, A gripper jaw comprising a jaw opening for receiving and holding the engagement section, A multichannel pipette according to claim 7 or 8, comprising: a gripping sleeve positioned around the gripping jaw, wherein the gripping sleeve is configured to exert an inward force on the gripping jaw such that the gripping jaw exerts an inward force on the engaging section of the plunger in order to hold the engaging section of the plunger within the gripping jaw when the locking mechanism is in the second locking configuration.

10. The multichannel pipette according to claim 9, wherein pressing the lock button causes the gripping sleeve to move distal to the gripper jaws, thereby applying the inward force to the gripper jaws.

11. The multichannel pipette according to claim 10, wherein by activating the third actuator, the gripping sleeve moves proximal to the gripper jaw, thereby retracting the inward force on the gripper jaw.

12. The multichannel pipette according to claim 11, wherein the first actuator comprises a button disposed at the proximal end of an elongated shaft, and the second actuator comprises a button disposed at the proximal end of an elongated shaft.

13. The multichannel pipette according to claim 12, wherein the lock button is disposed at the distal end of the shaft, the shaft of the lock button and the shaft of the second actuator are parallel, and when the second actuator moves from the first pressed position to the second unpressed position, or from the second unpressed position to the first pressed position, the lock button and the shaft of the lock button translate to the button of the second actuator and the shaft of the second actuator.

14. The multichannel pipette according to claim 13, wherein the locking mechanism comprises a push plate that reversibly contacts the proximal end of the gripping sleeve, and when the lock button is pressed, the push plate moves the gripping sleeve distally relative to the gripper jaw.

15. The multichannel pipette according to claim 14, wherein by activating the third actuator, the tip discharge sleeve moves distal to the gripper jaw and contacts the proximal end of the pipette tip arranged circumferentially around the gripper jaw, thereby disengaging the pipette tip.

16. The multichannel pipette according to claim 15, wherein the locking mechanism is in the second configuration, and while the locking mechanism is engaged with the engagement section of the plunger, the first actuator is moved from a first non-pressed position to a second pressed position, causing the plunger to move proximal to the gripper jaw, thereby drawing fluid into the pipette tip via the suction force generated by the proximal movement of the lumen section within the lumen of the pipette tip.

17. The multichannel pipette according to claim 16, wherein the locking mechanism is in the second configuration, and while the locking mechanism is engaged with the engagement section of the plunger, the second actuator is moved from the second non-pressed position to the first pressed position, thereby moving the plunger distally to the gripper jaw, and thereby dispensing fluid from the pipette tip via a displacement force generated by the distal movement of the lumen section within the lumen of the pipette tip.

18. The multichannel pipette according to claim 6, wherein the multichannel pipette comprises two, three, four, five, six, seven, or eight or more gripper mechanisms.

19. The multichannel pipette according to claim 18, wherein the multichannel pipette comprises eight gripper mechanisms.

20. A multichannel pipette according to any one of claims 1 to 19, further comprising a tip interface arranged circumferentially around each gripper jaw, the tip interface comprising a retaining platform configured to engage with tabs of a pipette tip retaining interface to secure the pipette tip to the pipette.

21. The multichannel pipette according to any one of claims 1 to 20, further comprising a tip ejection sleeve disposed on the tip interface, which, when the third actuator is activated, moves distally to the tip interface and operates to displace the pipette tip attachment interface, thereby removing the pipette tip from the pipette.

22. The multichannel pipette according to any one of claims 1 to 21, further comprising an electrode disposed in the distal section, wherein the electrode is electrically coupled to each gripper jaw of the plurality of gripper mechanisms.

23. The multichannel pipette according to claim 22, wherein each gripper jaw is made of a conductive material and is operable to allow an electrical pulse applied to the electrode to pass through the electrode, through each gripper jaw, through the plunger at the tip of the pipette, through the sample contained in the lumen of the tip of the pipette, and through a second electrode disposed adjacent to the distal end of the tip of the pipette, thereby enabling electroporation of cells contained in the sample.

24. It is an electroporation system, A multichannel pipette according to any one of claims 1 to 23, The tip of the pipette and Pipette docking assembly and An electroporation system equipped with a pulse generator.

25. The electroporation system according to claim 24, wherein the pipette docking assembly comprises a pipette station, a pipette station guard, and a reservoir.

26. The electroporation system according to claim 25, wherein the reservoir includes a buffer.

27. The electroporation system according to any one of claims 24 to 26, wherein the pipette tip is provided with a 10 μL pipette tip or a 100 μL pipette tip.

28. The electroporation system according to any one of claims 24 to 27, wherein the tip of the pipette comprises a plunger at least partially disposed within the lumen, and the plunger is configured to translate along the lumen to facilitate aspiration and / or dispensing.

29. The electroporation system according to claim 28, wherein the plunger comprises gold, diamond-like carbon, and / or conductive plastic.

30. The electroporation system according to claim 28 or 29, wherein the plunger comprises an engaging section and a lumen section, and the lumen section comprises a sealing component for creating a seal between the plunger and the lumen.

31. The electroporation system according to claim 30, wherein the lumen section comprises a front pin and a shaft section, the front pin being connected to the shaft section and configured to fix the sealing component to the shaft section.

32. The electroporation system according to claim 31, wherein the sealing component comprises a polymer sleeve, and the front pin is configured to be inserted through the polymer sleeve and engage with a retaining hole in the shaft section to secure the sealing component to the shaft section.

33. The electroporation system according to claim 32, wherein the insertion of the front pin through the polymer sleeve defines a space between the polymer sleeve and the front pin, and the space contributes to the flexibility of the polymer sleeve for creating the seal between the plunger and the lumen.

34. The electroporation system according to any one of claims 30 to 33, wherein the sealing component comprises polytetrafluoroethylene (PTFE).

35. The electroporation system according to claim 30, wherein the sealing component comprises a coated O-ring, and the lumen section includes a circumferential recess configured to receive the coated O-ring.

36. The electroporation system according to any one of claims 30 to 35, wherein the pipette tip further comprises a mounting interface adjacent to the lumen, the mounting interface comprising one or more tabs configured to engage with a retaining platform of the distal section of the pipette assembly.

37. The tip of the pipette is connected to the multichannel pipette and positioned in contact with the buffer within the reservoir. The first electrode is electrically coupled to the buffer, The plunger at the tip of the pipette is electrically in communication with the sample and pipette electrode inside the tip of the pipette. The electroporation system according to any one of claims 24 to 36, wherein the pipette electrode is in contact with a second electrode, and the first electrode and the second electrode are electrically in communication with the pulse generator.

38. The electroporation system according to any one of claims 24 to 37, 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 configured to operate as a power source for an amplifier circuit, and the amplifier circuit is configured to supply voltage to the sample in the multichannel pipette in a manner that takes into account load fluctuations.

39. The electroporation system according to claim 38, wherein the amplifier circuit comprises a common source amplifier configured to output high-voltage pulses.

40. The electroporation system according to claim 39, wherein the common source amplifier receives a signal from the amplitude setting loop, and the signal from the amplitude setting loop is based on an input from a digital-to-analog converter and an input from a voltage sensing loop, and the input from the voltage sensing loop is determined using the high-voltage pulse, voltage divider, and differential amplifier.

41. The electroporation system according to claim 40, wherein the common source amplifier amplifies the signal from the amplitude setting loop by a coefficient of approximately 1,000 to approximately 2,000.

42. The electroporation system according to claim 40 or 41, wherein the input from the digital-to-analog converter corresponds to a waveform selected by the user.

43. The electroporation system according to any one of claims 24 to 42, wherein the pulse generator comprises an arc discharge detection module configured to detect an arc discharge in the sample while a voltage is applied to the sample.

44. The aforementioned arc discharge detection module A first step 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, wherein the falling edge signal indicates a decrease in the current passing through the sample, and the decrease in current suggests an arc discharge. The electroporation system according to claim 43, further comprising: a comparator configured to determine whether an arc discharge has occurred in the sample by comparing the falling edge signal filtered by the bandpass filter with one or more reference standards.

45. The electroporation system according to claim 44, wherein the amplification of the current signal by the first step amplifier is based on the output of a low-voltage detection circuit for determining the resistance associated with the sample.

46. A method for transfecting cells with a payload, To provide an electroporation system according to any one of claims 24 to 45, To provide the aforementioned cells, To provide the aforementioned payload, The process involves introducing the aforementioned cells and payload into the tip of the pipette, A method comprising electroporating the cells by operating the electroporation system.

47. The method according to claim 46, wherein the cells are mammalian cells.

48. The method according to claim 46, wherein the cell is a microorganism or an organoid.

49. The method according to claim 46, wherein the payload is selected from the group consisting of nucleic acids, proteins, or combinations thereof.

Images