System for delivery of a payload into cell

Abstract

To provide a System for delivery of a payload into cell.SOLUTION: A system for delivering a payload to a cell that includes: a platform supporting an input container, an output container, and a receiver for receiving all or part of a disposable assembly, the disposable assembly including a preparation vessel and a constriction cartridge. The preparation vessel holds a cell suspension as it is prepared for passage through one or more cell-deforming constrictions, and the constriction cartridge houses a component that includes the one or more cell deforming constrictions. Passage through the cell-deforming constrictions causes perturbations in cell membranes to allow entry of a payload into the cells.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 62 / 608,202, filed December 20, 2017, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to techniques for the delivery of payloads into cells, and more particularly to a tabletop system for perturbing cell membranes to allow passage of a payload through the cell membrane. [Background technology]

[0003] The controlled delivery of various materials into cells is important in the developing medical field of cell therapy. For example, various research and therapeutic applications may involve the delivery of peptides, nucleic acids, proteins, small molecules, and nanomaterials through cell membranes and into cells. As discussed in WO2013059343, WO2015023982, PCT / US2015 / 058489, PCT / US2015 / 060689, and PCT / US2016 / 13113, constricting microfluidic channels can be used to deliver compounds and other payloads into cells. However, previous systems and methods for delivering materials into cells involve multiple separate instruments to prepare the cells and payload, pass the cells through the constriction, and process the cells following passage through the constriction. These multiple separate instruments may need to be operated by a variety of different technicians, and the overall cell processing procedure may be slowed down due to the time required to perform tasks by different people and with different instruments. Additionally, operation of multiple different instruments by different operators can lead to inconsistent results across different operators. Furthermore, various numbers of separate instruments can occupy scarce and expensive space and time in a laboratory clean room. Summary of the Invention [Means for solving the problem]

[0004] As mentioned above, known methods for intracellular payload delivery utilize many different types of laboratory equipment, many different operators performing spatially and temporally distributed tasks, and large amounts of space and time in laboratory clean rooms. Therefore, improved techniques for intracellular payload delivery are needed. In some embodiments, the disclosed systems, methods, and techniques for intracellular delivery can be performed at a tabletop scale by a single operator on a single piece of laboratory equipment suitable for use inside a laboratory clean room. The systems, methods, and techniques disclosed herein can improve the processing time, throughput rate, consistency, and efficiency of intracellular payload delivery processes at a clinical scale and in a clinical setting.

[0005] In some embodiments, a tabletop laboratory and / or clinical system is provided, whereby the tabletop laboratory and / or clinical device is configured to accept a cell suspension and a payload for delivery of the cell suspension into cells and to force the cell suspension through a disposable constriction cartridge to create a perturbation in the membrane of the cells in the cell suspension. The system may be configured to automatically control the flow, temperature, agitation, and / or pressure of the cell suspension and / or its environment before, during, and after the system flows the cell suspension through the constriction cartridge. In some embodiments, the system may be controllable by a single user operating a user interface to control the flow, temperature, agitation, pressure, pH, concentration, and / or other characteristics and properties of the cell suspension and / or its environment. In some embodiments, the system includes one or more disposable components configured for single use, which may be attached to the system by hand, without the use of tools, such that the cell suspension may flow through one or more of the disposable components before, during, and / or after constriction of the cells.

[0006] In some embodiments, a first system for delivering a payload to cells is provided, the system including: a holder configured to hold a cell suspension input container containing a cell suspension comprising cells; a receiver configured to receive all or a portion of a disposable assembly, the disposable assembly being in fluid communication with the input container and configured to hold the cell suspension as it is prepared for passage through one or more cell deformation constrictions; and a constriction cartridge configured to be in fluid communication with the preparation container, the constriction cartridge comprising one or more cell deformation constrictions configured to store components that cause perturbations in the plasma membranes of the cells. and a receiver comprising a constriction cartridge configured to allow entry of a payload into the cells; and one or more processors configured to receive input from a user and control one or more control modules, the one or more control modules configured to control one or more of the pressure, temperature, agitation, and flow of the cell suspension, the one or more control modules comprising a flow control module configured to flow the cell suspension from the input container, through the disposable assembly, to a cell suspension output container so that the payload is delivered into the cells.

[0007] In some embodiments, a first disposable assembly is provided for use in a system for delivering a payload to cells, the first disposable assembly comprising: a preparation vessel configured to hold a cell suspension as it is prepared for passage through one or more cell-deforming constrictions; and a constriction cartridge configured to be in fluid communication with the preparation vessel, the constriction cartridge configured to store components comprising one or more cell-deforming constrictions, the cell-deforming constrictions configured to create a perturbation in a cell membrane that allows entry of the payload into the cell.

[0008] In some embodiments, a first method for delivering a payload to cells is provided, the first method including the steps of providing cells in a cell suspension, passing the cell suspension into a preparation vessel in a tabletop system, preparing the cell suspension, including by applying pressure to the cell suspension while the cell suspension is in the preparation vessel, and passing the prepared cell suspension from the preparation vessel through a constriction cartridge of the system, the constriction cartridge configured to store components comprising a cell-deforming constriction that creates a perturbation in the membrane of the cell that allows entry of the payload into the cell.

[0009] In some embodiments, a second system for delivering a payload through a cell membrane is provided, the second system comprising: a preparation vessel configured to contain a cell suspension, the suspension comprising cells; a constriction cartridge fluidly connected to the preparation vessel; a touchscreen display; one or more processors; and a memory configured to store instructions executable by the one or more processors to cause the system to detect a contact on the display at a location corresponding to an icon for initiating a process for delivering a payload through the membrane of cells in the cell suspension, and in accordance with detecting the contact, adjust a temperature of the cell suspension inside the preparation vessel, apply pressure to the cell suspension inside the preparation vessel, and pass the cell suspension from the preparation vessel through a constriction in a component contained within the constriction cartridge, the constriction being a cell-deforming constriction that creates a perturbation in the membrane of cells in the cell suspension that allows entry of the payload into the cells.

[0010] In some embodiments, a third system for delivering a payload to cells is provided, the third system including: a holder configured to hold a cell suspension input container containing a cell suspension comprising cells; a receiver configured to receive a disposable assembly, the disposable assembly in fluid communication with the input container and configured to hold the cell suspension as it is prepared for passage through one or more cell deformation constrictions; and a constriction cartridge configured to be in fluid communication with the preparation container, the constriction cartridge comprising one or more cell deformation constrictions configured to store components that create perturbations in the plasma membrane of the cells to allow entry of the payload into the cells. a receiver comprising a constriction cartridge configured to fluidly connect the cell suspension to the input container, a cell suspension output container configured to be in fluid communication with the constriction cartridge, and one or more processors configured to receive input from a user and control one or more control modules configured to control one or more of pressure, temperature, agitation, and flow of the cell suspension, the one or more control modules comprising a flow control module configured to flow the cell suspension from the input container, through the disposable assembly, and to the output container such that a payload is delivered into the cells.

[0011] In some embodiments, a second disposable assembly is provided for use in a system for delivering a payload to cells, the second disposable assembly comprising: a preparation vessel configured to hold a cell suspension as it is prepared for passage through one or more cell deformation constrictions; a constriction cartridge configured to be in fluid communication with the preparation vessel, the constriction cartridge configured to store components comprising one or more cell deformation constrictions, the cell deformation constrictions configured to create a perturbation in a cell membrane that allows entry of the payload into the cells; and a cell suspension output container configured to be in fluid communication with the constriction cartridge.

[0012] In some embodiments, a second method for delivering a payload to cells is provided, the second method including providing the cells in a cell suspension; passing the cell suspension into a preparation vessel in a tabletop system; preparing the cell suspension, including by applying pressure to the cell suspension while the cell suspension is in the preparation vessel; and passing the prepared cell suspension from the preparation vessel through a constriction cartridge of the system, the constriction cartridge configured to store components comprising a cell-deforming constriction that creates a perturbation in the membrane of the cell that allows entry of the payload into the cell.

[0013] In some embodiments, a fourth system for delivering a payload through a cell membrane is provided, the fourth system comprising: a preparation vessel configured to contain a cell suspension, the suspension comprising cells; a constriction cartridge fluidly connected to the preparation vessel; a touchscreen display; one or more processors; and a memory configured to store instructions executable by the one or more processors to cause the system to detect a contact on the display at a location corresponding to an icon for initiating a process for delivering a payload through the membrane of cells in the cell suspension, and, in accordance with detecting the contact, adjust the temperature of the cell suspension inside the preparation vessel, apply pressure to the cell suspension inside the preparation vessel, and pass the cell suspension from the preparation vessel through a constriction in a component contained within the constriction cartridge, the constriction being a cell-deforming constriction that creates a perturbation in the membrane of cells in the cell suspension that allows entry of the payload into the cells.

[0014] In some embodiments, any one or more of the features, characteristics, or elements discussed above with respect to any of the system, method, or assembly embodiments may be incorporated into any of the other system, method, or assembly embodiments described above. In some embodiments, any one or more of the features, characteristics, or elements discussed anywhere in this disclosure may be incorporated into any of the system, method, or assembly embodiments described above. The present invention provides, for example, the following. (Item 1) 1. A system for delivering a payload to a cell, the system comprising: A platform, a holder configured to hold a cell suspension input container containing a cell suspension comprising cells; A receptacle configured to receive all or a portion of a disposable assembly, the disposable assembly comprising: a preparation vessel in fluid communication with the input container and configured to hold the cell suspension as it is prepared for passage through one or more cell-deforming constrictions; a constriction cartridge configured to be in fluid communication with the preparation vessel, the constriction cartridge configured to house a component comprising the one or more cell-deforming constrictions, the cell-deforming constrictions configured to create a perturbation in a cell membrane of the cell to allow entry of a payload into the cell; a receiver comprising: one or more processors configured to receive input from a user and control one or more control modules configured to control one or more of pressure, temperature, agitation, and flow of the cell suspension, the one or more control modules comprising: a flow control module configured to flow the cell suspension from the input container, through the disposable assembly, and to a cell suspension output container such that the payload is delivered into the cells. one or more processors, Supporting platform A system comprising: (Item 2) the component comprising the one or more cell deformation constrictions is a microfluidic chip comprising a microfluidic channel, the microfluidic channel comprising the one or more cell deformation constrictions; the constriction cartridge is a microfluidic chip cartridge configured to store the microfluidic chip; Item 1. The system of item 1. (Item 3) the component comprising the one or more cell deformation constrictions is a filter comprising a plurality of cell deformation constrictions; the constriction cartridge is a filter cartridge configured to house the filter; 3. The system according to any one of items 1 and 2. (Item 4) 4. The system of any one of items 1-3, wherein the platform is a tabletop platform. (Item 5) 5. The system of any one of items 1-4, wherein the cell suspension comprises the payload. (Item 6) 6. The system of any one of items 1-5, wherein the system is configured to contact the payload with the cell suspension prior to flow of at least a portion of the cell suspension through the constriction cartridge. (Item 7) 7. The system of any one of items 1-6, wherein the system is configured to contact the payload with the cell suspension following flow of at least a portion of the cell suspension through the constriction cartridge. (Item 8) 8. The system of any one of items 1-7, wherein the one or more processors are configured to apply pressure to the cell suspension inside the preparation vessel. (Item 9) Item 10. The system of item 8, wherein the one or more control modules include a pressure control module including a pressure source configured to deliver sterilizing gas to the preparation vessel. Item 11. The system according to any one of items 8 and 9, wherein the pressure applied to the cell suspension inside the preparation vessel is sufficient to bring the preparation vessel into contact with the inner wall of the receiving vessel. 11. The system of any one of items 8-10, wherein the pressure applied to the cell suspension inside the preparation vessel is sufficient to force the cell suspension out of the preparation vessel through the constriction cartridge. (Item 12) Item 13. The system of any one of items 1-11, wherein the one or more processors are configured to heat or cool the cell suspension inside the preparation vessel. Item 13. The system of item 12, wherein the one or more control modules comprise a temperature control module comprising one or more thermoelectric temperature control devices configured to heat or cool a portion of the receiver configured to contact the preparation vessel. (Item 14) Item 15. The system of item 13, wherein the portion of the receiver configured to contact the preparation vessel is a conductive jacket configured to conduct heat to and from the preparation vessel. 15. The system of any one of items 12-14, wherein the one or more control modules comprise a temperature control module comprising one or more thermoelectric temperature control devices configured to heat or cool the preparation vessel. (Item 16) Item 16. The system of item 15, wherein the one or more thermoelectric temperature control devices comprise a cooling plate, the cooling plate being disposed within a wall of the receptacle and configured to contact an outer wall of the preparation vessel when the preparation vessel is inserted into the receptacle. (Item 17) 17. The system of any one of items 1-16, wherein the one or more processors are configured to agitate the cell suspension inside the preparation vessel so that cells are homogenously dispersed in the cell suspension. (Item 18) Item 18. The system of item 17, wherein the one or more control modules comprise an agitation control module comprising a stir plate configured to be driven by one or more motors, the stir plate configured to cause agitation of all or a portion of the receiver. (Item 19) 19. The system of any one of items 1-18, wherein the flow control module is configured to cause one or more valves to control the flow of the cell suspension from the input container, through the disposable assembly, and to the output container. (Item 20) 20. The system of any one of items 1-19, wherein the flow control module is configured to flow the cell suspension at a target fluid velocity. (Item 21) 21. The system of any one of items 1-20, further comprising an input device configured to receive instructions from the user, wherein the one or more processors are configured to operate one or more of the control modules in response to the instructions. (Item 22) Item 22. The system of item 21, wherein the input device comprises a touchscreen display configured to transmit a signal to one or more of the control modules in response to detecting contact by a user. (Item 23) 23. The system of any one of items 21 and 22, wherein the input device comprises an agitation speed adjustment device configured to control the speed of a motor causing agitation of the cell suspension inside the preparation vessel. (Item 24) 24. The system of any one of items 1-23, wherein the preparation vessel is configured to hold up to 10 liters of the cell suspension. (Item 25) 25. The system of any one of items 1-24, wherein the preparation vessel is configured to allow a pressure of up to 120 psi to be applied to the cell suspension. (Item 26) 26. The system of claim 1, wherein the preparation vessel is configured to allow a pressure of up to 120 psi to be applied to the interior of the preparation vessel. (Item 27) 27. The system of any one of items 1-26, wherein the preparation vessel comprises a first inlet, the first inlet being fluidly connected to the cell suspension input container and configured to receive a flow of the cell suspension. (Item 28) 28. The system of any one of items 1-27, wherein the preparation vessel comprises a second inlet, the second inlet being fluidly connected to a pressure source and configured to receive a flow of sterilizing gas into the preparation vessel to apply pressure to the cell suspension. (Item 29) 29. The system of any one of items 1-28, wherein the preparation vessel comprises an outlet configured to be fluidly connected to the constriction cartridge. (Item 30) 30. The system of any one of items 1-29, wherein the constriction cartridge comprises an inlet configured to be fluidly connected to the preparation vessel. (Item 31) 31. The system of any one of items 1-30, wherein the constriction cartridge comprises a housing portion configured to hold the component comprising the one or more cell deforming constrictions and direct flow of the cell suspension through the component comprising the one or more cell deforming constrictions. (Item 32) 32. The system of any one of items 1-31, wherein the constriction cartridge is configured such that the components are held between a base portion and a removable lid portion. (Item 33) Item 33. The system of item 32, wherein the removable lid portion is configured to be slidably attachable to and removable from the base portion without the use of tools. (Item 34) Item 35. The system of any one of items 1-33, wherein the constriction cartridge is configured to hold the components in place with one or more O-rings. 35. The system of any one of items 1-34, wherein the constriction cartridge is configured to direct the flow of the cell suspension into the component through one or more O-rings. (Item 36) the disposable assembly comprising a cell suspension output container configured to be in fluid communication with the constriction cartridge; the constriction cartridge comprises an outlet configured to be fluidly connected to the output container. Item 36. The system of any one of items 1-35. (Item 37) 37. The system of any one of items 1-36, wherein the one or more processors are configured to receive signals from one or more sensors and automatically control one or more of pressure, temperature, and agitation of the cell suspension according to the received signals. (Item 38) Item 38. The system of item 37, wherein the one or more sensors are contained within the disposable assembly. (Item 39) 39. The system of any one of items 37 and 38, wherein the one or more sensors comprise a temperature sensor configured to monitor the temperature of the cell suspension. (Item 40) 40. The system of claim 39, wherein the temperature sensor comprises a thermistor, the thermistor being included in the disposable assembly and configured to be attached to the receptacle. (Item 41) 41. The system of any one of items 37-40, wherein the one or more sensors comprise a bubble sensor configured to monitor the flow of the cell suspension. (Item 42) Item 43. The system of any one of items 37-41, wherein the one or more sensors include a pressure sensor configured to monitor a pressure applied to the cell suspension. 43. The system of any one of items 37-42, wherein the one or more sensors comprise a pressure sensor configured to monitor the pressure inside the preparation vessel. (Item 44) 44. The system of any one of items 1-43, further comprising a memory configured to store log information comprising one or more of pressure, temperature, agitation, flow, and elapsed time while the cell suspension is located within one or both of the preparation vessel and the constriction cartridge. (Item 45) 45. The system of any one of items 1-44, further comprising a display configured to display information comprising one or more of pressure, temperature, agitation, flow, and elapsed time while the cell suspension is located within one or both of the preparation vessel and the constriction cartridge. (Item 46) 46. ​​The system of any one of items 1-45, further comprising a network communication interface configured to transmit information to a remote computing device, the information comprising one or more of pressure, temperature, agitation, flow, and elapsed time while the cell suspension is located within one or both of the preparation vessel and the constriction cartridge. (Item 47) 47. The system of any one of items 1-46, wherein the system is configured to be capable of being moved from a first location to a second location without disassembly. (Item 48) Item 48. The system of item 47, wherein moving the system without disassembly includes moving the platform without removing the holder, receiver, or one or more processors from the platform. (Item 49) Item 49. The system of any one of items 1-48, wherein the system is less than 2 feet in height. (Item 50) 50. The system of any one of items 1-49, wherein the system has a footprint of less than 3 feet by 2 feet. (Item 51) The system of any one of items 1-50, wherein the system is less than 60 pounds. (Item 52) 52. The system of any one of items 1-51, wherein the system is configured to be sterilizable. (Item 53) 53. The system of any one of items 1-52, further comprising a filter configured to receive fluid flow downstream from the preparation vessel and upstream of the constriction cartridge, the filter configured to remove multicellular aggregates from the cell suspension before reaching the constriction cartridge. (Item 54) Item 54. The system of item 53, wherein the filter is configured to withstand an internal pressure of greater than 120 psi. (Item 55) 55. The system of any one of items 1-54, wherein the preparation container comprises a flexible plastic bag. (Item 56) Item 56. The system of item 55, wherein the receptacle comprises a housing configured to receive the flexible plastic bag so that the contents of the bag can be cooled. (Item 57) 57. The system of any one of items 1-56, wherein the preparation vessel comprises a circulation loop configured to draw liquid from a main body of the preparation vessel and circulate the liquid back into the main body of the preparation vessel. (Item 58) Item 58. The system of item 57, wherein the circulation loop comprises a peristaltic pump configured to cause the liquid to flow through the circulation loop. (Item 59) 59. The system of any one of items 57 and 58, wherein part of the circulation loop is integrated with a flow path leading from the preparation vessel to the constriction cartridge. (Item 60) 1. A disposable assembly for use in a system for delivering a payload to a cell, said assembly comprising: a preparation vessel configured to hold a cell suspension as it is prepared for passage through one or more cell-deforming constrictions; a constriction cartridge configured to be in fluid communication with the preparation vessel, the constriction cartridge configured to house a component comprising the one or more cell-deforming constrictions, the cell-deforming constrictions configured to create a perturbation in a cell membrane that allows entry of a payload into the cell; An assembly comprising: (Item 61) Item 61. The assembly of item 60, wherein the assembly is configured to be capable of being connected to and disconnected from the system without the use of tools. (Item 62) the component comprising the one or more cell deformation constrictions is a microfluidic chip comprising a microfluidic channel, the microfluidic channel comprising the one or more cell deformation constrictions; the constriction cartridge is a microfluidic chip cartridge configured to store the microfluidic chip; 62. The assembly according to any one of items 60 and 61. (Item 63) the component comprising the one or more cell deformation constrictions is a filter comprising a plurality of cell deformation constrictions; the constriction cartridge is a filter cartridge configured to house the filter; 63. The assembly according to any one of items 60-62. (Item 64) 64. The assembly of any one of items 60-63, wherein the system is a tabletop system. (Item 65) 65. The assembly of any one of items 60-64, wherein connecting the assembly to the system without the use of a tool comprises fluidly connecting the assembly to the system such that the system can receive a flow of the cell suspension from the system. (Item 66) 66. The assembly of any one of items 60-65, wherein connecting the assembly to the system without the use of a tool comprises electronically connecting one or more sensors of the assembly to one or more controllers of the system. (Item 67) Item 67. The assembly of item 66, wherein the one or more sensors comprise a temperature sensor configured to monitor the temperature of the cell suspension inside the preparation vessel and transmit data regarding the temperature to a temperature control module of the system. (Item 68) 68. The assembly of any one of items 66 and 67, wherein the one or more sensors comprise a pressure sensor configured to monitor the pressure applied to the cell suspension and transmit data regarding the pressure to a pressure control module of the system. (Item 69) 69. The assembly of any one of items 66-68, wherein the pressure sensor is configured to monitor the pressure inside the preparation vessel and transmit data regarding the pressure to a pressure control module of the system. (Item 70) 70. The assembly of any one of items 66-69, wherein the one or more sensors comprise a bubble sensor configured to monitor the flow of the cell suspension through the assembly. (Item 71) 71. The assembly of any one of items 60-70, wherein the constriction cartridge comprises an inlet configured to be fluidly connected to the preparation vessel. (Item 72) 72. The assembly of any one of items 60-71, wherein the constriction cartridge comprises a housing portion configured to hold the component comprising the one or more cell deforming constrictions and to direct flow of the cell suspension through the component comprising the one or more cell deforming constrictions. (Item 73) 73. The assembly of any one of items 60-72, wherein the constriction cartridge is configured such that the components are held between a base portion and a removable lid portion. (Item 74) Item 74. The assembly of item 73, wherein the removable lid portion is configured to be slidably attachable to and removable from the base portion. (Item 75) 75. The assembly of any one of items 60-74, wherein the constriction cartridge is configured to hold the components in place by one or more O-rings. (Item 76) 76. The assembly of any one of items 60-75, wherein the constriction cartridge is configured to direct the flow of the cell suspension into the component through one or more O-rings. (Item 77) 77. The assembly of any one of items 60-76, further comprising a cell suspension output container configured to be in fluid communication with the outlet of the constriction cartridge. (Item 78) 78. The assembly of any one of items 60-77, wherein the constriction cartridge houses a second component comprising one or more cell-deforming constrictions and is configured to direct the flow of the cell suspension fluid through the second component in parallel with the first component. (Item 79) 79. The assembly of any one of items 60-78, wherein the assembly is configured to be sterilizable. (Item 80) 1. A method for delivering a payload to a cell, the method comprising: providing cells in a cell suspension; passing the cell suspension into a preparation vessel in a tabletop system; preparing the cell suspension, including by applying pressure to the cell suspension while the cell suspension is in the preparation vessel; passing the prepared cell suspension from the preparation vessel through a constriction cartridge of the system, the constriction cartridge configured to store a component comprising a cell-deforming constriction that creates a perturbation in the membrane of the cells to allow entry of a payload into the cells; A method comprising: (Item 81) the component comprising the one or more cell deformation constrictions is a microfluidic chip comprising a microfluidic channel, the microfluidic channel comprising the one or more cell deformation constrictions; the constriction cartridge is a microfluidic chip cartridge configured to store the microfluidic chip; Item 80. The method according to item 80. (Item 82) the component comprising the one or more cell deformation constrictions is a filter comprising a plurality of cell deformation constrictions; the constriction cartridge is a filter cartridge configured to house the filter; 82. The method according to any one of items 80 and 81. (Item 83) 83. The method of any one of items 80-82, further comprising contacting the payload with the cell suspension prior to flowing at least a portion of the cell suspension through the constriction cartridge. (Item 84) 84. The method of any one of items 80-83, further comprising contacting the payload with the cell suspension following flow of at least a portion of the cell suspension through the constriction cartridge. (Item 85) 85. The method of any one of items 80-84, further comprising attaching to the system a disposable assembly comprising the preparation vessel and the constriction cartridge. (Item 86) Item 86. The method of item 85, wherein attaching the disposable assembly includes attaching the disposable assembly without the use of a tool. (Item 87) 87. The method of any one of items 85 and 86, wherein attaching the disposable assembly comprises inserting the preparation vessel into a receptacle of the system configured to receive the preparation vessel. (Item 88) 88. The method of any one of items 85-87, wherein attaching the disposable assembly includes attaching one or more sensors contained within the disposable assembly such that the sensors are configured to transmit signals to the system. (Item 89) Item 90. The method of item 88, wherein the one or more sensors included in the disposable assembly include a temperature sensor configured to monitor the temperature of the cell suspension. Item 90. The method of item 89, wherein the temperature sensor comprises a temperature probe configured to be attached to the receptacle. (Item 91) 91. The method of any one of items 88-90, wherein the one or more sensors included in the disposable assembly comprises a bubble sensor configured to monitor the flow of the cell suspension. (Item 92) 92. The method of any one of items 88-91, wherein the one or more sensors included in the disposable assembly comprise a pressure sensor configured to monitor pressure applied to the cell suspension. (Item 93) 93. The method of any one of items 85-92, wherein attaching the disposable assembly includes attaching one or more sensors included in the system to the disposable assembly, wherein the sensors are configured to detect one or more characteristics related to the contents of the disposable assembly. (Item 94) Item 94. The method of item 93, wherein the one or more sensors included in the system comprise a flow sensor configured to monitor the flow of liquid through tubing of the disposable assembly. (Item 95) 95. The method of any one of items 93 and 94, wherein the one or more sensors included in the system comprise a level sensor configured to monitor the fill level of the preparation vessel. (Item 96) 96. The method of any one of items 93-95, wherein the one or more sensors included in the system comprise a temperature sensor configured to monitor the temperature of the liquid in the preparation vessel. (Item 97) 97. The method of any one of items 80-96, wherein preparing the cell suspension comprises stirring the cell suspension inside the preparation vessel to homogenously disperse the cells in the cell suspension. (Item 98) 98. The method of claim 97, wherein agitating the cell suspension comprises monitoring the agitation of the cell suspension. (Item 99) 99. The method of any one of items 80-98, wherein preparing the cell suspension comprises heating or cooling the cell suspension inside the preparation vessel. (Item 100) 100. The method of claim 99, wherein heating or cooling the cell suspension comprises using a thermoelectric temperature control device to heat or cool a portion of a receiver containing the preparation vessel. (Item 101) 101. The method of any one of items 99 and 100, wherein heating or cooling the cell suspension comprises monitoring the temperature of the cell suspension as it is heated or cooled. (Item 102) 102. The method of any one of items 80-101, wherein applying pressure to the cell suspension comprises monitoring pressure as it is applied to the cell suspension. (Item 103) 103. The method of any one of items 80-102, wherein providing the cells in a cell suspension comprises providing the cell suspension in an input container. (Item 104) 104. The method of any one of items 80-103, comprising passing the cell suspension from the constriction cartridge into an output container. (Item 105) 105. The method of any one of items 80-104, comprising performing an integrity check on the disposable assembly prior to passing the cell suspension through the disposable assembly. (Item 106) Item 106. The method of item 105, wherein performing an integrity check on the disposable assembly includes pressurizing a gas inside the disposable assembly and monitoring the pressure of the pressurized gas over a predetermined period of time. (Item 107) 107. The method of any one of items 80-106, comprising passing a primer solution through the disposable assembly prior to passing the cell suspension through the disposable assembly. (Item 108) providing a supply of blood comprising a first plurality of types of cells; isolating cells of a target type from among said first plurality of cells of a type; suspending said isolated target-type cells in a delivery material to form said cell suspension; 8. The method according to any one of items 80-107, further comprising: (Item 109) washing the treated cells following passage through the cell deformation constriction to remove the delivery material; suspending the washed treated cells in a buffer for cryopreservation; Item 109. The method of item 108, further comprising: (Item 110) monitoring the fill level of the preparation vessel using one or more level sensors while all or part of the cell suspension is in the preparation vessel; 109. The method of any one of items 80-109, further comprising: (Item 111) circulating the cell suspension out of the main body of the preparation vessel through a circulation loop and back into the main body of the preparation vessel while the cell suspension is in the preparation vessel. 111. The method of any one of items 80-110, further comprising: (Item 112) and ceasing circulation of the cell suspension through the circulation loop in response to a determination that the fill level of the preparation vessel is below a predetermined threshold. Item 112. The method of item 111, further comprising: (Item 113) 1. A system for delivering a payload across a cell membrane, said system comprising: a preparation vessel configured to contain a cell suspension, the suspension comprising cells; and a constriction cartridge fluidly connected to the preparation vessel; A touchscreen display and one or more processors; a memory configured to store instructions executable by the one or more processors, the instructions providing the system with: detecting a contact on the display at a location corresponding to an icon to initiate a process for delivering a payload through membranes of cells in the cell suspension; In response to detecting the contact, adjusting the temperature of the cell suspension inside the preparation vessel; applying pressure to the cell suspension inside the preparation vessel; passing the cell suspension from the preparation vessel through a constriction in a component contained within the constriction cartridge, the constriction being a cell-deforming constriction that creates a perturbation in the membrane of cells in the cell suspension that allows entry of the payload into the cells; memory and A system comprising: (Item 114) The instructions are executable by the one or more processors, and provide the system with: While controlling the temperature of the suspension inside the preparation vessel, displaying an indication of the current pressure being applied to the cell suspension; displaying an indication of the current temperature of the cell suspension; displaying a dynamic indication of elapsed time for the payload delivery process; and Item 114. The system according to Item 113, (Item 115) The instructions are executable by the one or more processors, and the system includes: While passing the cell suspension through the constriction cartridge, continuing to display an indication of the current pressure being applied to the cell suspension; continuing to display an indication of the current temperature of the cell suspension; continuing to display a dynamic indication of elapsed time for the payload delivery process; and displaying a dynamic indication of the elapsed time for the process of passage of the cell suspension through the constriction cartridge; and Item 115. The system according to item 114, (Item 116) The instructions are executable by the one or more processors, and the system includes: Detecting that the payload delivery process is complete; and In response to detecting that the payload delivery process is completed, ceasing to display a dynamic indication of elapsed time for the payload delivery process; and stopping the display of a dynamic indication of the elapsed time for the process of passage of the cell suspension through the constriction cartridge; and displaying an indication of the total elapsed time for the payload delivery process; and displaying an indication of the total elapsed time for the process of passage of the cell suspension through the constriction cartridge; and Item 116. The system according to item 115, (Item 117) Item 118. The system of any one of items 113-116, wherein heating or cooling the cell suspension inside the preparation vessel is performed by detecting a touch on the display at a location corresponding to an icon for performing a heating or cooling process. 118. The system of any one of items 113-117, wherein applying pressure to the cell suspension inside the preparation vessel comprises delivering pressurized gas into the preparation vessel at a pressure indicated by one or more contacts detected on the display at locations corresponding to icons for setting pressure. (Item 119) 119. The system of any one of items 113-118, wherein applying pressure to the cell suspension comprises delivering pressurized gas into the preparation vessel at a pressure indicated by one or more contacts detected on the display at locations corresponding to icons for setting pressure. (Item 120) The instructions are executable by the one or more processors to cause the system to agitate the cell suspension inside the preparation vessel; agitating the cell suspension inside the preparation vessel is performed according to detecting a contact on the display at a location corresponding to an icon for performing an agitation process. 119. The system of any one of items 113-119. (Item 121) The instructions are executable by the one or more processors, and the system includes: displaying instructions for connecting a disposable assembly in response to detecting that the disposable assembly is not connected to the system; and ceasing the display of instructions to connect to the disposable assembly in response to detecting that the disposable assembly is connected to the system; The system according to any one of items 113-120, (Item 122) The instructions are executable by the one or more processors, and the system includes: In accordance with detecting that a first portion of the disposable assembly is connected, ceasing the display of a first page of instructions, the first page comprising instructions for connecting a first portion of the disposable assembly; and replacing the display of the first page of instructions with a display of a second page of instructions, the second page comprising instructions for connecting to a second portion of the disposable assembly; Item 122. The system according to Item 121, (Item 123) The instructions are executable by the one or more processors, and the system includes: Detecting a touch on the display at a location corresponding to an icon for initiating an integrity test; initiating the integrity test in response to detecting the contact and displaying an indication of the current pressure and an indication of the elapsed time for the integrity test; The system of any one of items 113-122, (Item 124) The instructions are executable by the one or more processors, and provide the system with: detecting a touch on the display at a location corresponding to an icon for initiating a priming process; In response to detecting the contact, passing a primer solution through the preparation vessel and the constriction cartridge; displaying the current temperature of the primer solution during the priming process; 124. The system of any one of items 113-123, [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 illustrates a tabletop system for delivering a payload to a cell, according to some embodiments.

[0016] [Figure 2] FIG. 2 illustrates a partially see-through view of a pressure control module of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0017] [Figure 3A] FIG. 3A illustrates a platform of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0018] [Figure 3B] FIG. 3B illustrates a detailed view of a platform of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0019] [Figure 4] FIG. 4 illustrates a partially see-through view of a temperature control module of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0020] [Figure 5A] FIG. 5A illustrates a preparation vessel housing of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0021] [Figure 5B] FIG. 5B illustrates an exploded view of a door of a preparation vessel housing of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0022] [Figure 6A] FIG. 6A illustrates a preparation vessel of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0023] [Figure 6B] FIG. 6B illustrates a cap of a preparation vessel of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0024] [Figure 7A] FIG. 7A illustrates an exploded view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to house two tips.

[0025] [Figure 7B] FIG. 7B illustrates a side cross-sectional view and detailed extract of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store two chips.

[0026] [Figure 7C]FIG. 7C illustrates a partially transparent overhead view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to house two chips.

[0027] [Figure 8A] FIG. 8A illustrates a first view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store four tips.

[0028] [Figure 8B] FIG. 8B illustrates a second view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store four tips.

[0029] [Figure 8C] FIG. 8C illustrates a first partially exploded view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store four tips.

[0030] [Figure 8D] FIG. 8D illustrates a second partially exploded view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store four tips.

[0031] [Figure 9A] FIG. 9A illustrates a partially assembled sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0032] [Figure 9B]FIG. 9B illustrates a first detailed view of a sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0033] [Figure 9C] FIG. 9C illustrates a second detailed view of a sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0034] [Figure 9D] FIG. 9D illustrates a sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0035] [Figure 10] FIG. 10 illustrates a schematic diagram of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0036] [Figure 11A] 11A and 11B illustrate schematic diagrams of systems for providing pressurized gas for use in delivering a payload to a cell, according to some embodiments. [Figure 11B] 11A and 11B illustrate schematic diagrams of systems for providing pressurized gas for use in delivering a payload to a cell, according to some embodiments.

[0037] [Figure 12] FIG. 12 illustrates a method for treating cells, including intracellular payload delivery, according to some embodiments.

[0038] [Figure 13] FIG. 13 illustrates a method for intracellular payload delivery, according to some embodiments.

[0039] [Figure 14A]14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14B] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14C] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14D] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14E] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14F] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14G] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14H] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14I] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14J] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14K] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14L] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14M] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14N] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14O] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14P] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14Q] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14R] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14S] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14T] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14U] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 14V] 14A-14V illustrate a user interface for controlling a tabletop system for delivering a payload to a cell, according to some embodiments.

[0040] [Figure 15A] 15A-15C illustrate a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 15B] 15A-15C illustrate a tabletop system for delivering a payload to a cell, according to some embodiments. [Figure 15C] 15A-15C illustrate a tabletop system for delivering a payload to a cell, according to some embodiments.

[0041] [Figure 16] FIG. 16 illustrates a schematic of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0042] [Figure 17] FIG. 17 illustrates a flexible bag for holding cell suspension fluid as it is prepared for passage through a constriction component of a tabletop system for delivering a payload to cells, according to some embodiments.

[0043] [Figure 18] FIG. 18 illustrates a flexible bag for holding cell suspension fluid as it is prepared for passage through a constriction component of a tabletop system for delivering a payload to cells, according to some embodiments.

[0044] [Figure 19A]19A-19D illustrate a flexible bag for holding cell suspension fluid during the performance of four different functions of a tabletop system for delivering a payload to cells, according to some embodiments. [Figure 19B] 19A-19D illustrate a flexible bag for holding cell suspension fluid during the performance of four different functions of a tabletop system for delivering a payload to cells, according to some embodiments. [Figure 19C] 19A-19D illustrate a flexible bag for holding cell suspension fluid during the performance of four different functions of a tabletop system for delivering a payload to cells, according to some embodiments. [Figure 19D] 19A-19D illustrate a flexible bag for holding cell suspension fluid during the performance of four different functions of a tabletop system for delivering a payload to cells, according to some embodiments.

[0045] [Figure 20] FIG. 20 is a computer, according to some embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0046] Described below are exemplary embodiments of tabletop laboratory and / or clinical systems for partially or fully automated intracellular payload delivery, as well as associated devices, systems, methods, techniques, and user interfaces. Below, the description of Figures 1-11B primarily describes exemplary embodiments of tabletop systems and associated devices for intracellular payload delivery, which systems and devices may be used in conjunction with the methods, techniques, and user interfaces described herein. Thereafter, the description of Figures 12 and 13 primarily describes methods and techniques for cell processing and intracellular payload delivery, which may be implemented by or in conjunction with the systems, methods, devices, and user interfaces described elsewhere herein. Next, the description of Figures 14A-14V primarily describes exemplary user interfaces for controlling the systems and devices and for implementing the methods and techniques for intracellular payload delivery. Next, Figures 15-19D primarily describe further exemplary embodiments of tabletop laboratory and / or clinical systems and associated devices for intracellular payload delivery, including flexible bags usable therein, which systems and devices may be used in conjunction with the methods, techniques, and user interfaces described herein. Finally, the description of Figure 20 primarily describes a computing device that may be integrated into or used in conjunction with any of the systems or devices described herein.

[0047] The following description describes example systems, methods, techniques, parameters, and the like, however, it should be recognized that such description is not intended as a limitation on the scope of the present disclosure, but is instead provided as a description of example embodiments. definition

[0048] For purposes of interpreting this specification, the following definitions shall apply, and where appropriate, terms used in the singular shall also include the plural and vice versa. In the event that any definition set forth below conflicts with any document incorporated herein by reference, the set forth definition shall control.

[0049] As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated.

[0050] It should be understood that the aspects and embodiments of the invention described herein include "comprising," "consisting," and "consisting essentially of" aspects and embodiments.

[0051] Furthermore, it will be understood that the terms "includes," "including," "comprises," and / or "comprising" specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups.

[0052] The term "if" may be interpreted to mean "when" or "upon" or "in response to determining" or "in response to detecting," depending on the context. Similarly, the phrase "when it is determined" or "when [a stated condition or event] is detected" may be interpreted to mean "in response to determining" or "in response to determining," or "in response to detecting [a stated condition or event]," or "in response to detecting [a stated condition or event]," depending on the context.

[0053] The term "about," as used herein, refers to a normal error range for a particular value that is readily ascertained by one of ordinary skill in the art. As used herein, reference to "about" a value or parameter includes (and describes) embodiments that are directed to the value or parameter itself.

[0054] Although the description herein uses the terms "first," "second," etc. to describe various elements, these elements should not be limited by the terms. These terms are used only to distinguish one element from another.

[0055] With respect to any of the structural and functional properties described herein, methods for determining these properties are known in the art.

[0056] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety. Intracellular payload delivery systems and devices

[0057] As described below, the techniques, systems, and methods disclosed herein can provide intracellular payload delivery that can be performed at a tabletop scale by a single operator in a single laboratory instrument suitable for use inside a laboratory clean room. Furthermore, the techniques, systems, and methods disclosed herein can provide intracellular payload delivery systems with removable and / or disposable components, such as components that can be configured for one-time use, that can be quickly and conveniently attached to the overall system by hand, without the use of tools, in a closed, sterile environment. As described below, the disposable components of the systems described herein may be configured for use in a sterile environment and yet be sufficiently robust to withstand the pressures used to force fluids to flow through the system. The systems, methods, and techniques disclosed herein can significantly improve the processing time, throughput rate, consistency, and efficiency of intracellular payload delivery processes. Below, Figures 1-20 provide descriptions of exemplary techniques, systems, and methods for intracellular payload delivery, according to some embodiments.

[0058] 1 illustrates a tabletop system 100 for delivering a payload to cells, according to some embodiments. System 100 may be a tabletop system, such as a piece of laboratory equipment, configured to receive a cell suspension fluid, process the cell suspension fluid, and deliver a payload to cells in the cell suspension. System 100 may be configured to be operated by a single user, two users, or more than two users, such as by being controlled by an electronic user interface, in some embodiments, presented via one or more buttons, keyboards, keypads, dials, knobs, and / or touchscreen interfaces.

[0059] In some embodiments, system 100 may include one or more computing devices configured to enable fully or partially automated control of one or more components of system 100. As described below, system 100 may include one or more control modules, such as a pressure control module and a temperature control module, all of which may contain one or more computing devices. As used herein, the term “control module” may refer to one or more components of a system that work together to control one or more properties of the system and / or of a fluid or other medium controlled by the system, such as a pressure control module that controls pressure or a temperature control module that controls temperature. In some embodiments, a control module may comprise one or more physical components (e.g., sensors, control devices such as pumps, or heating devices, etc.) and / or one or more electronic devices (e.g., computer processor, computer memory, etc.). In some embodiments, a control module may refer to multiple system components that may or may not be physically separated / isolated from other system components and that may or may not be removable from the system. In some embodiments, all or a portion of a control module may be provided in a single housing with all or a portion of another control module. In some embodiments, a single system component (e.g., a processor, a sensor, a pump, a portion of tubing, etc.) may form part of only one control module or of more than one control module. Computing devices as described and referenced herein may comprise one or more processors coupled to a computer-readable storage medium that stores computer instructions that, when executed, may cause the one or more computing devices to perform all or a portion of one or more of the methods described herein.In some embodiments, one or more of the computing devices described herein may be electronically / communicatively coupled to each other to send and receive electronic signals representing data, information, and / or instructions. In some embodiments, one or more of the computing devices described herein may be coupled to one or more electronic devices configured to send signals to one or more computing devices (e.g., sensors) and / or receive instructions from one or more computing devices (e.g., electronically controllable valves). The electronic communication coupling of the computing devices and associated electronic components may, in some embodiments, include wired electronic communication via a wired network or computer bus, and in some embodiments, wireless electronic communication via a wireless computer network. The various systems described herein, including systems 100, 1000, 1100, 1500, and 1600 and various associated electronic devices, may all include one or more computing devices as described above. Thus, this disclosure may describe certain actions (e.g., generating data, detecting input, controlling electronic components) performed by a system or by a component or device of a system, and it will be understood by one of ordinary skill in the art that these actions may be performed by one or more computing devices of the system or device. Where specific hardware or software is not specifically described to perform an action performed by any of the systems or devices described herein, it will be understood by one of ordinary skill in the art that the action may be performed by a computing device of the system or device.

[0060] In some embodiments, as described in further detail below, system 100 may be configured for use inside a sterile environment (e.g., a closed sterile environment), such as a laboratory clean room. For example, system 100 may be configured such that all of its components (including disposable components, as described below) may be sterile (e.g., pre-sterilized) and / or sterilizable so as not to contaminate the sterile environment. In addition, one or more components of system 100 may be made from a material acceptable for use in a sterile environment, such as stainless steel (e.g., marine grade stainless steel, 316 grade stainless steel, 316L grade stainless steel, etc.). In addition, system 100 may include one or more particulate filters to ensure compliance with requirements for operation within a sterile environment. Furthermore, system 100 may be made compact so as not to occupy scarce and expensive space inside a sterile environment, where small spaces are often at a premium. For example, in some embodiments, system 100 may be less than about 3 feet, less than about 2 feet, or less than about 1 foot in length. In some embodiments, system 100 may be greater than about 2 feet, greater than about 1 foot, or greater than about 6 inches in length. In some embodiments, the system may be less than about 2 feet, less than about 1 foot, or less than about 8 inches deep. In some embodiments, the system may be greater than about 1 foot, greater than about 8 inches, or greater than about 4 inches deep. In some embodiments, the system may be less than about 3 feet, less than about 2 feet, or less than about 1 foot in height. In some embodiments, the system may be greater than about 2 feet, greater than about 1 foot, or greater than about 6 inches in height. In some embodiments, system 100 may weigh less than about 60 pounds, less than about 40 pounds, less than about 20 pounds, or less than about 10 pounds. In some embodiments, system 100 may weigh more than about 40 pounds, more than about 20 pounds, more than about 10 pounds, or more than about 5 pounds.

[0061] As shown in FIG. 1 , system 100 may be a tabletop system with several components mounted on platform 102. Platform 102 may be a rigid base configured to support the weight of the other components of system 100 and configured so that the other components of system 100 can be securely mounted to platform 102 in fixed locations. Platform 102 may be configured to sit on a tabletop, laboratory bench, mobile cart, or the like. In some embodiments, platform 102 may be configured to sit on the floor. By constructing the various components of system 100 so that they are all supported by platform 102, system 100 may be made compact, lightweight, and portable. For example, system 100 may be sufficiently lightweight and compact so that platform 102 can be moved or transported from one laboratory space to another, and the other components can move with platform 102, such that system 100 can be fully functional immediately after platform 102 is moved from one location to another.

[0062] System 100 may further include a hook 104, which may be configured to suspend one or more bags. In the example shown in FIG. 1 , hook 104 is configured to suspend a cell suspension input bag 106 and a buffer input bag 108. Bags 106 and 108 may be flexible bags configured to contain liquids to be passed through and processed by system 100. Bags 106 and 108 may be suspended from hook 104 and configured to be fluidly connected to flow paths of system 100. The flow paths, which will be described in more detail throughout this specification, may be flow paths through which the cell suspension is configured to process the cell suspension and prepare the cells of the cell suspension for payload delivery, and in some embodiments, to effect payload delivery. In some embodiments, the flow paths may originate in bags 106 and 108 and lead through one or more pipes or flexible tubing into a preparation vessel housed within preparation vessel housing 110.

[0063] System 100 may further include a preparation vessel housing 110, which may be any structure or component configured to store a preparation vessel. As described in more detail below, the preparation vessel may be any vessel configured to store a cell suspension fluid as it is prepared for passage through a constriction component that defines a portion of a flow path configured to create a perturbation within the membrane of cells in the cell suspension fluid to facilitate entry of a payload into the cells through the membrane. For example, the preparation vessel may be configured to hold a cell suspension while the suspension is cooled (or heated) and agitated as air pressure is applied thereto and / or as the cell suspension is otherwise manipulated or controlled to be forced through the constriction component. Preparation vessel housing 110, in some embodiments, may be a rigid housing, such as the rectangular housing shown in FIG. 1, and may have an inlet opening for the flow path to enter the housing and an outlet opening for the flow path to exit the housing. In the embodiment of FIG. 1, the inlet opening is at the top of the housing 110 and the outlet opening is at the bottom of the housing 110 so that the flow path into, through, and out of the preparation vessel can be gravity-assisted.

[0064] In some embodiments, the housing 110 may be configured to be able to open and close so that a preparation vessel inside the housing (and / or other components inside the housing) can be inserted, adjusted, and / or removed. In the example of FIG. 1 , the housing 110 has a hinged door on the front that can be opened and closed. In some embodiments, the housing 110 may be configured to contact the preparation vessel (e.g., when the preparation vessel is inserted into the housing) to facilitate one or more preparation processes. For example, the housing 110 may be configured to contact the preparation vessel and facilitate heat transfer between the preparation vessel and the housing 110, and / or the housing 110 may be configured to contact the preparation vessel and facilitate agitation of the cell suspension by shaking the housing 110 and transferring motion to the preparation vessel.

[0065] In some embodiments, in addition to the preparation vessel, the housing 110 may be further configured to store one or more additional components, such as components defining additional portions of the flow path. For example, as discussed in more detail below, the housing 110 may additionally be configured to store the entire disposable assembly including the preparation vessel and the constriction cartridge, which may be configured to store components, such as a microfluidic chip or filter, that define a portion of the flow path configured to produce a cell membrane perturbation. In some embodiments, the preparation vessel may additionally be configured to store and / or be connected to one or more sensors, such as a temperature sensor (e.g., a thermistor), a flow sensor (e.g., a bubble sensor, a flow rate sensor), a pressure sensor, a weight sensor, an accelerometer, a CO2 sensor, a pH sensor, an osmometer, a conductivity sensor, an impedance detector, an optical-based detector, and / or any one or more sensors configured to measure cell concentration, membrane disruption, and / or other properties of the cells or of the cell suspension. Any of the sensors listed herein may, in some embodiments, be connected to a preparation vessel, in some embodiments may be connected to other system components, and / or may be configured to be in direct contact with fluid flowing through the system (e.g., inside the preparation vessel or otherwise inside the flow path of the system).

[0066] In some embodiments, in addition to the temperature sensors discussed below with reference to sensor assembly 900, housing 110 may include one or more integrated temperature sensors configured to detect the temperature of a portion of the housing and transmit data to the system according to the detected temperature. In some embodiments, the one or more integrated temperature sensors may be configured for use in determining the temperature of the housing, the preparation vessel, and / or the cell suspension inside the preparation vessel, as discussed further below.

[0067] System 100 may further include an output bag tray 111, which may be a tray or other holder component configured to hold, store, or otherwise support one or more bags or other containers configured to receive fluid after flow through the flow paths of system 100. In the example of FIG. 1 , output bag tray 111 is configured to hold a cell suspension output bag 112 and a buffer output bag 114. Output bags 112 and 114 may share one or more properties in common with input bags 106 and 108, but output bags 112 and 114 may be configured to begin the intracellular payload delivery process empty and end the process full. That is, output bags 112 and 114 may be filled with cell suspension and buffer fluid, respectively, after fluid flow through the flow paths of system 100. In some embodiments, output bags 112 and 114 may be fluidly connected to a flow path through an outlet opening at the bottom of preparation vessel housing 110 such that fluid may flow through the preparation vessel and constriction cartridge, and then out of housing 110 into one of bags 112 and 114. In some embodiments, such as the example shown in FIG. 1 , output bag tray 111 may be positioned on platform 102 such that the output bags it supports are located near and / or below preparation vessel housing 110, and fluid flow from housing 110 toward tray 111 may be gravity-assisted.

[0068] System 100 may further include a pressure control module 116, which may comprise any structure, housing, or component of system 100 configured to store pressure control hardware and / or software, as will be described in more detail below. In the example of FIG. 1 , pressure control module 116 is mounted on platform 102 and comprises a rigid structure housing a touchscreen display 120, which may be configured to display a graphical user interface for controlling one or more operations of system 100. In some embodiments, pressure control module 116 may be electrically and mechanically coupled to one or more other components of system 100, such that a processor located within pressure control module 116 may send signals to and receive signals from other electronic components of system 100, and such that pressure control hardware (e.g., pumps, filters, etc.) may be fluidly connected to deliver and receive one or more pressurized fluids (e.g., sterilant gas, air, etc.) to and from the other components of system 100.

[0069] System 100 may further include a temperature control module 118, which may comprise any structure, housing, or component of system 100 configured to store temperature control hardware and / or software, as will be described in more detail below. In the example of FIG. 1, temperature control module 118 comprises a rigid structure mounted on platform 102. In some embodiments, temperature control module 118 may be electrically and mechanically coupled to one or more other components of system 100, such that a processor located within temperature control module 118 may send and receive signals to and from other electronic components of system 100, and such that temperature control hardware (e.g., heating / cooling elements) may be physically connected to transfer heat to or from other components of system 100.

[0070] 2 illustrates a partially transparent view of a pressure control module 116 of a tabletop system 100 for delivering a payload to cells, according to some embodiments. In some embodiments, the pressure control module 116 is the same pressure control module 116 as discussed above with reference to FIG. 1. In some embodiments, the pressure control module 116 may receive a flow of pressurized gas from an external source, such as a pressurized canister, adjust the pressure of the flow, direct the flow through one or more filters, and direct the flow into a preparation vessel of the system, so that the pressurized gas may force a flow of cell suspension through a constriction cartridge of the system. In some embodiments, the pressure control module 116 may be configured to be attachable to one or more filter assemblies, which may be removable and / or replaceable, and in some embodiments, may be configured to be attachable by one or more inlets or outlets external to the body / housing of the pressure control module.

[0071] 2, pressure control module 116 may include a touchscreen display 120 mounted on an exterior surface of the housing of module 116. Touchscreen display 120 may be electronically connected to other electronic components of system 100 for sending and receiving control and display signals. Touchscreen display 120 may be configured to display a graphical user interface for controlling one or more operations of system 100. In some embodiments, rather than a touchscreen display, pressure control module 120 may include a non-touchscreen display, one or more buttons, one or more knobs, one or more sliders, one or more keyboards, one or more additional touchscreen or non-touchscreen displays, and / or any other component configured to receive input from a user for controlling the operation of system 100. In some embodiments, touchscreen display 120 (or other user controls, such as those described above) may be located on another component of system 100 and may likewise be electronically coupled to a component of system 100.

[0072] The pressure control module 116 may further comprise various internal components, as described below. The pressure control module 116 may comprise an electro-pneumatic regulator assembly 204 and may be configured to control the flow of gas using one or more valves (e.g., push valves, vent valves, solenoid valves 208, etc.) to maintain an output gas pressure at a pressure input or otherwise indicated by a user or by the system. The electro-pneumatic regulator 204 may be controlled by one or more integrated or external processors. The electro-pneumatic regulator 204 may be configured to inhale gas from a sterilizing gas source or from the environment (e.g., to inhale air) and output pressurized gas. The electro-pneumatic regulator 204 may be configured to be fluidly connected to an output path for pressurized gas, which may be fluidly connected to an opening in the preparation vessel, as discussed above. In some embodiments, the opening in the preparation vessel configured to receive the flow of pressurized gas may be located on or near the preparation vessel. In this manner, the electro-pneumatic regulator 204 may be configured to deliver pressurized gas to a preparation vessel above the cell suspension fluid (or buffer fluid) such that the pressurized gas applies pressure downward on the fluid.

[0073] The pressure control module 116 may further include a PID controller 202, which may be any proportional-integral-derivative controller, configured to regulate the pressure. The PID controller 202 may be electronically coupled to the electro-pneumatic regulator 204 and may be configured to receive a signal indicative of the current pressure and deliver a control signal to the electro-pneumatic regulator 204 based on calculations made based on the current pressure, the control signal being configured to maintain the desired pressure produced by the electro-pneumatic regulator 204 in an optimal manner (e.g., without using excessive power and without causing pressure corrections that unnecessarily exceed the desired pressure).

[0074] Pressure control module 116 may further include an air filter (e.g., a gas filter) and a regulator assembly 206, which may include a pressure regulator and an air filter. In some embodiments, air filter and regulator assembly 206 may be configured to be attached to a source of pressurized gas, such as a pressurized canister containing gas (e.g., air, nitrogen, etc.). The pressure regulator of assembly 206 may be fluidly connected to the source of pressurized gas and may be configured to reduce the input pressure to a desired pressure, such as a pressure that may be set by a user or by the system. The pressure regulator may further be fluidly connected to the air filter of assembly 206, which may be an optional filter configured to remove contaminants from the gas flowing from the regulator. Thus, gas may flow from the pressurized gas source, through the regulator, then through the air filter, and then finally to and into electro-pneumatic regulator 204, as described above.

[0075] The pressure control module 116 may further include an electronics board 212 that may be electronically connected to any one or more electronic components of the module 116 (or other electronic components of the system 100) and configured to send and receive signals to and from the components.

[0076] The pressure control module 116 may further include a power supply and relay assembly 210 that may be configured to provide power to one or more components of the pressure control module 116 (such as any of the components discussed above) and / or any other electrically powered components.

[0077] 3A and 3B illustrate a platform 102 of a tabletop system 100 for delivering a payload to a cell, according to some embodiments. In some embodiments, the platform 102 is the same platform 102 discussed above with reference to FIG. 1.

[0078] As shown in FIG. 3A , the platform 102 may have a motor 302 and a shaker plate 304 mounted on the platform's upper surface. The motor 302 may be configured to drive a belt, such as shown as part of a belt drive 306 in FIG. 3B , to cause movement of the shaker plate 304. In some embodiments, the motor 302 may be connected to the shaker plate 304 by one or more belts, drive axles, or other mechanical components. In the embodiment of FIGS. 3A and 3B , the motor 302 and the shaker plate 304 are connected by the belt drive 306, which is located on the underside of the platform 102. However, in some embodiments, the motor 302 and the shaker plate 304 may be connected by one or more components located above, near, and / or within the platform 102.

[0079] In some embodiments, the shaker plate 304 may be configured to support a preparation vessel housing, such as the preparation vessel housing 110, as discussed above with respect to Figure 1. Movement of the shaker plate 304 (e.g., oscillating, circular, back-and-forth, and / or up-and-down) may cause agitation of the cell suspension located inside a preparation vessel housed within the preparation vessel housing when the housing is mounted on the shaker plate 304. Thus, movement of the shaker plate 304 may be configured to cause agitation of the cell suspension to prevent cells from falling out of suspension, as discussed further below.

[0080] In some embodiments, the drive speed of the motor 302 may be controllable by one or more input devices of the system 100, such as a physical input knob and / or a touchscreen user interface.

[0081] 4 illustrates a partially see-through view of a temperature control module 118 of a tabletop system 100 for delivering a payload to a cell, according to some embodiments. In some embodiments, the temperature control module 118 is the same temperature control module 118 as discussed above with reference to FIG.

[0082] 4, the temperature control module 118 may be mounted on the platform 102 near the preparation vessel housing 110 and the output bag tray 111. In some embodiments, the temperature control module 118 may be mounted such that it is in proximity to and / or in physical contact with the preparation vessel housing 110 and can be easily electrically and / or physically connected to the preparation vessel housing 110, as discussed above.

[0083] The temperature control module 118 may further comprise various internal components, as described below. In some embodiments, the temperature control module 118 may include one or more components configured to heat and / or cool the preparation vessel housing 110 or its components. For example, the temperature control module 118 may include one or more forced air heaters, one or more forced air coolers, one or more thermoelectric cooling devices (e.g., Peltier coolers), one or more resistive heating devices, one or more liquid heating devices, one or more liquid cooling devices, or the like. In some embodiments, the heating and / or cooling components may be in electronic, electrical, and / or physical contact with the preparation vessel housing 110. In some embodiments, one or more Peltier coolers, such as a stainless steel jacket forming the interior wall of the housing 110 and configured to contact the preparation vessel 600, may be used to cool a coolant that may be circulated to contact all or a portion of the housing 110 and draw heat away therefrom.

[0084] The temperature control module 118 may include a vent gas input 402. In some embodiments, the vent gas input 402 may be configured to allow excess gas to be vented from one or more regulators of the system (such as any of the regulators discussed below) so that the gas may be vented to the interior of the system, thereby preventing pressure buildup within the system, limiting vibrations, and preventing particle contamination within the sterile environment.

[0085] Temperature control module 118 may further include filter 404, which may be any filter configured to pass exhaust air generated by the system through before being released into the environment outside system 100. For example, filter 404 may be a HEPA filter, such that system 100 may be suitable for use in a sterile environment, as the exhaust air is passed through a HEPA filter before being released into the sterile environment.

[0086] 4 also shows mounts 408 for hooks 104. Mounts 408 may be mounted on platform 102, the housing of pressure control module 116, and / or the housing of temperature control module 118. Mounts 408 may be configured such that they can be loosened and / or tightened to adjust the position of hooks 104.

[0087] 5A and 5B illustrate a preparation vessel housing 110 of a tabletop system 100 for delivering a payload to a cell, according to some embodiments. In some embodiments, the preparation vessel housing 110 is the same preparation vessel housing 110 as discussed above with reference to FIG. 1.

[0088] As shown in FIG. 5A , preparation vessel housing 110 may be a rigid housing with a rectangular exterior configured to house preparation vessel 600 and constriction cartridge 700, both of which will be described in more detail below. As described above, preparation vessel 600 and constriction cartridge 700 may each define a portion of a flow path for the cell suspension and may be connected to each other by tubing. As shown in FIG. 5A , preparation vessel housing 110 may include inlet opening 512 and outlet opening 514. The openings may be positioned and configured such that piping or tubing defining the cell suspension flow path of system 100 can pass therethrough. In some embodiments, inlet opening 512 is located at or near the top of housing 110, while outlet opening 514 is located at or near the bottom of housing 110, such that fluid flow through the flow path traveling from inlet opening 512 to outlet opening 514 can be gravity-assisted. In the embodiment shown, the inlet opening 512 is configured to allow buffer fluid or cell suspension fluid to enter through a tube and air for pressurization of the preparation vessel 600 to enter through another tube. In the embodiment shown, the outlet opening 514 is configured to allow buffer fluid or cell suspension fluid to exit through a tube after passing through the preparation vessel 600 and the constriction cartridge 700.

[0089] As shown in FIG. 5A, the inner opening of the preparation vessel housing 110 may be shaped so that the preparation vessel 600 and constriction cartridge 700 fit securely into place within the housing 110 and contact the inner walls of the housing 110.

[0090] The preparation vessel housing 110 may further include a door 504, as shown in FIG. 5B , which may allow access to the interior of the housing 110. In the example of FIGS. 5A and 5B , the door 504 is hinged along one side and closeable by a latch 510 along the other side. As shown in FIG. 5B , a window 506 in the door 504 may be acted upon by a spring 508. The spring 508 may, in some embodiments, attach the window 506 to the door 504 and be arranged such that they are compressed when the door 504 is closed and latched with the preparation vessel 600 inside the housing 110. For example, when the door 504 is closed, the window 506 may press against the preparation vessel 600, compressing the spring 508. The spring force of the spring 508 may thus press the window 506 firmly against the preparation vessel 600, ensuring that the preparation vessel 600 is held firmly in place against the interior wall of the housing 110.

[0091] In some embodiments, one or more interior walls of the housing 110 may be configured to contact an exterior surface of the preparation vessel 600 such that the walls may transfer heat to and / or from the preparation vessel 600. In some embodiments, the interior walls of the housing 110 may be made from a metal, such as stainless steel, to facilitate fast and efficient transfer of heat from the walls to the preparation vessel 600. In some embodiments, the spring force of the spring 508 may press the window 506 firmly against the preparation vessel 600, ensuring that the preparation vessel 600 is in contact with the interior walls of the housing 110, thereby facilitating optimal heat transfer.

[0092] 6A and 6B illustrate a preparation vessel 600 of tabletop system 100 for delivering a payload to cells, according to some embodiments. In some embodiments, preparation vessel 600 is the same preparation vessel 600 as discussed above with reference to FIGS. 5A and 5B and / or the same preparation vessel as discussed above with reference to FIG. 1. Preparation vessel 600 may be any vessel or container configured to store fluids to be passed through the system, including cell suspension fluid, buffer fluid, and / or pressurized gas. That is, preparation vessel 600 may be configured to hold cell suspension fluid while the cell suspension is prepared for passage through a constriction cartridge, such as constriction cartridge 700. In some embodiments, preparing the cell suspension for passage through a constriction cartridge may include cooling the suspension, heating the suspension, agitating the suspension, and / or applying pressure to the suspension.

[0093] As shown in FIG. 6A , the preparation vessel 600 may be a rigid syringe-shaped container in some embodiments. In some embodiments, the preparation vessel 600 may be made of plastic or other suitable inert and sterilizable material. In some embodiments, the preparation vessel 600 may have a volume of about 25 mL, about 50 mL, about 100 mL, about 250 mL, about 500 mL, about 1 L, about 2 L, about 5 L, or about 10 L. In some embodiments, the preparation vessel 600 may have a volume of more than about 10 mL, 25 mL, about 50 mL, about 100 mL, about 250 mL, about 500 mL, about 1 L, about 2 L, about 5 L, or about 10 L. In some embodiments, the preparation vessel 600 may have a volume of less than about 25 mL, about 50 mL, about 100 mL, about 250 mL, about 500 mL, about 1 L, about 2 L, about 5 L, about 10 L, or about 20 L. In some embodiments, preparation vessel 600 may be formed from a modified 250-mL medical-grade syringe.

[0094] In some embodiments, preparation vessel 600 may have a tapered shape at its bottom portion to gravity-direct fluid flow within the vessel toward vessel outlet 606, which may be located at or near the bottom end of preparation vessel 600. In some embodiments, vessel inlet 604 may be located at or near the top end of preparation vessel 600. In the embodiment shown in FIG. 6A , vessel inlet 604 is an opening formed in vessel cap 608, a removable cap configured to seal the top opening of vessel 600 and be held in place by one or more O-rings 610, which may be medical-grade O-rings. In some embodiments, vessel 600 may be provided without a removable cap, or the removable cap or other lid or door may be held in place by a mechanism other than an O-ring, such as by threads, a clamp, an adhesive, or the like.

[0095] In some embodiments, one or more of openings 604 and 606 may be configured to fluidly connect to tubing, plumbing, or other system components that may define flow paths for cell suspension fluid, gas, or both. In some embodiments, openings 604 and / or 606 may include threads, Luer taper connectors, Luer lock connectors, Luer slip connectors, slip tip connectors, or other connector mechanisms to allow connection to other components.

[0096] In some embodiments, the preparation vessel 600 may be configured to allow the gas inside the preparation vessel to be pressurized to at least the operating pressure of the system 100. In some embodiments, the operating pressure of the system 100 may be air pressure against which the gas inside the preparation vessel 600 is pressurized to force the cell suspension fluid from the preparation vessel 600, out the vessel outlet 606, and through the constriction cartridge 700. In some embodiments, the operating pressure of the system 100 may be about 20 psi, about 30 psi, about 50 psi, about 70 psi, about 90 psi, about 110 psi, or about 130 psi. In some embodiments, the operating pressure may be greater than 10 psi, 20 psi, 50 psi, 70 psi, 90 psi, 110 psi, or 130 psi. In some embodiments, the operating pressure may be less than 20 psi, 50 psi, 70 psi, 90 psi, 110 psi, 130 psi, or 150 psi. Preparation vessel 600 may be constructed and configured such that its cap, openings, valves, and other components may remain intact under operating pressures.

[0097] 7A-7C illustrate various views of a constriction cartridge 700 of a tabletop system 100 for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store two tips. In some embodiments, the constriction cartridge 700 is the same constriction cartridge 700 as discussed above with reference to FIGS. 1, 5A, and 5B. FIG. 7A illustrates an exploded view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store two tips. FIG. 7B illustrates a side cross-sectional view and a detailed extract view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store two tips. FIG. 7C illustrates a partially transparent overhead view of a constriction cartridge of a tabletop system for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store two tips.

[0098] In some embodiments, the constriction cartridge 700 may be any structure configured to contain or store a narrowing component, such as a narrowing filter (containing one or more narrowing microfluidic pores) or a narrowing microfluidic chip (containing one or more narrowing microfluidic channels). (Narrowing filters, according to some embodiments, are disclosed in Application No. WO / 2017 / 041050A1, which is incorporated herein by reference in its entirety.) Note that in some embodiments, a narrowing microfluidic channel or a narrowing microfluidic pore may simply be referred to as a "constriction" or a "cell-deforming constriction." The narrowing component may be any component having a channel, passage, or other opening (e.g., a constriction) with a diameter smaller than the cells of the cell suspension, such that forcing the cells through the opening under pressure creates a perturbation in the membrane of the cells as they are constricted by the opening. In some embodiments, constriction cartridge 700 may include an integrated constriction filter or microfluidic channel configured to constrict cells, while in some embodiments, constriction cartridge 700 may itself be configured to house a distinct component, including a constriction filter or a constriction microfluidic channel. In either case, constriction cartridge 700 may define a flow path of a portion of system 100 such that a cell suspension may flow from preparation vessel 600 toward and into constriction cartridge 700, and such that the cell suspension may then flow through and out of flow constriction cartridge 700 toward and into output bag 112 or 114 (or other suitable downstream flow path component).

[0099] 7A-7C , constriction cartridge 700 is disposed on cartridge body 702 and includes a constriction cartridge inlet 708 and a constriction cartridge outlet 710 that define the beginning and end of flow paths for the cell suspension and buffer fluids flowing through constriction cartridge 700. In some embodiments, constriction cartridge inlet 708 and constriction cartridge outlet 710 may include any one or more connection mechanisms discussed above with respect to the inlet and outlet of preparation vessel 600, such as threaded connections and / or Luer-type connections. Between constriction cartridge inlet 708 and constriction cartridge outlet 710, the flow path of system 100 may diverge into two or more parallel portions as fluids progress through constriction cartridge 700 and then reconverge before flowing out of constriction cartridge 700. In some embodiments, rather than defining multiple parallel flow paths through separate constriction components, constriction cartridge 700 may instead force fluid to flow serially through multiple constriction components, one after the other. In some embodiments, constriction cartridge 700 may be configured to be capable of accepting an empty surrogate component in place of a functional constriction component, which may not contain any channels or pores or may otherwise be configured to prevent flow through the portion of constriction cartridge 700 that houses the surrogate component. By using an empty surrogate component, constriction cartridge 700 may force fluid flow through only one constriction component at a time, such that the system need not be used with two constriction components at all times.

[0100] 7A-7C, constriction cartridge 700 is configured to flow the cell suspension (and buffer fluid) into and through constriction component 706, which may be a constriction microfluidic chip having multiple constriction microfluidic channels, or a constriction filter having multiple constriction openings or pores. In either case, constriction component 706 may have a separate constriction component inlet 716 for fluid flow into the component and a separate constriction cartridge outlet 718 for fluid flow out of the component.

[0101] Both the constriction component inlet 716 and the constriction component outlet 718 may be positioned to align with one of the O-ring fluid connections 712, which may be part of the cartridge body 702 configured to house an O-ring 714 and force fluid flow through the O-ring to and / or from the constriction component 706. The O-ring 714 may create a seal such that fluid may flow through the connection 712 and proceed between the body 702 and the constriction component 706 without leaking out of the flow path defined by the O-ring. In some embodiments, other sealing options other than, or in addition to, O-rings may be used to create a seal for the fluid connection between the constriction cartridge and the constriction component. For example, overmolding, chemical bonding, and / or mechanical interlocking may be used.

[0102] As shown in the detailed view of Figure 7B, the flow path inside the cartridge body 702 may diverge into multiple path portions flowing toward the O-ring fluid connector, and may reconverge following multiple path portions flowing from the O-ring fluid connector. In the example of Figure 7B, the flow path diverges from one path into two portions at a T-junction, causing fluid to flow from the cartridge inlet 708 into both constriction components 706, and the two portions converge into one path at the T-junction, causing fluid to flow from both constriction components 706 toward the cartridge outlet 710.

[0103] Thus, a fluid, such as a buffer fluid or a cell suspension, may flow into constriction cartridge 700 via constriction cartridge inlet 708, then toward and through an upstream pair of O-ring fluid connections 712. The fluid may flow through the upstream pair of O-ring fluid connections 712 into each constriction component inlet 716. From constriction component inlet 716, the fluid may flow through one or more channels or flow paths defined by constriction component 706 and then out of constriction component 706 at each of constriction component outlets 718. From constriction component outlet 718, the fluid may flow through a downstream pair of O-ring fluid connections and reconverge to flow toward and out of constriction cartridge outlet 710, thereby flowing out of constriction cartridge 700. Thus, in short, a fluid such as a buffer fluid or a cell suspension can flow into the constriction cartridge 700 and can be passed through one or more constriction components before flowing out of the constriction cartridge 700.

[0104] As shown in FIGS. 7A-7C , the constriction cartridge 700 may include a removable cover 704, which may be an element configured to be placed over the constriction components 706 and urge the constriction components 706 toward the cartridge body 702. In some embodiments, the removable cover 704 may be configured to apply an inward force to the constriction components 706, urging them toward the cartridge body 702 using one or more springs or other compressible components, such as rubber O-rings. In some embodiments, the removable cover 704 may be configured to be urged flush against a surface of one of the constriction components 706. In some embodiments, the removable cover 704 may serve to ensure that the constriction components 706 do not delaminate under the pressure of fluid being forced therethrough. That is, by holding the top of the constriction component 706 downward under force, the constriction component 706 may be prevented from delaminating. 7A-7C may provide superior durability under pressure over other mechanisms that may be used to manually assemble a constriction cartridge, such as attaching a cover with threaded components. In some embodiments, in addition to or as an alternative to one or more removable covers, a constriction cartridge may be configured to securely store constriction components without the use of a removable cover.

[0105] In some embodiments, the removable cover 704 may be configured to be removable by a user, such as by a hinge mechanism, a catch mechanism, a sliding mechanism, a threading mechanism, a locking mechanism, or other attachment and detachment mechanism. In the example shown in FIG. 7A , the removable cover 704 may be slid laterally to attach and detach the cartridge body 702 so that the constriction component 706 and O-ring 714 may be adjusted and / or replaced. As shown in FIG. 7A , interlocking tooth elements on the removable cover 704 and the body 702 may be configured to slide together and hold the removable cover 704 in place.

[0106] 8A-8D illustrate various views of a constriction cartridge 800 of a tabletop system 100 for delivering a payload to a cell, according to some embodiments, where the constriction cartridge is configured to store four tips. In some embodiments, the constriction cartridge 800 may share some or all characteristics in common with the constriction cartridge 700 as discussed above with reference to FIGS. 1, 5A, 5B, and / or 7A-7C, but the constriction cartridge may be configured to hold four constriction components rather than two.

[0107] In general, the constriction cartridge of system 100 may be configured to hold any number of constriction components for use in parallel (or, alternatively, in series) by configuring the shape of the body of the cartridge (and removable cover) to support the desired number of constriction components. For example, a two-constriction component cartridge may have a planar body as shown in FIGS. 7A-7C, a three-constriction component cartridge may have a triangular body, a four-constriction component cartridge may have a rectangular body as shown in FIGS. 8A-8D, etc. A cartridge body having any given number of sides and configured to hold a given number of constriction components may have a single inlet and a single outlet, but rather than a T-shaped junction where the flow path diverges into two path segments (and a corresponding T-shaped junction where the two path segments converge into one), the constriction cartridge body may instead have a junction where the flow path diverges into a given number of path segments, one path segment going toward each of the body's faces.

[0108] 8A-8D, constriction cartridge 800 has a body 802 and a removable cover 804, which may share any one or more characteristics in common with body 702 and removable cover 704, respectively, as described above with respect to Figures 7A-7C. As shown, body 802 may have an inlet 808 and an outlet 810, which may share any one or more characteristics in common with inlet 708 and outlet 710, respectively, as described above with respect to Figures 7A-7C.

[0109] As shown, body 802 may have four sides configured to support constriction component 806, which may each share any one or more characteristics in common with constriction component 706 as described above with respect to FIGS. 7A-7C. Removable cover 804, in some embodiments, may be configured to be removable from body 802 by sliding it upward or downward, as shown in FIG. 8C. Unlike removable cover 704 in FIGS. 7A-7C, removable cover 804 may not have teeth for attaching to the constriction cartridge body, as in some embodiments, removable cover 804 may be configured to completely surround the constriction cartridge body, as shown, thereby preventing lateral movement once slid into place.

[0110] 8D , constriction cartridge 800 may further include an O-ring fluid connection 812 disposed within body 802, which may share any one or more characteristics in common with O-ring fluid connection 712 and O-ring 714, respectively, as described above with respect to FIGS. 7A-7C , along with an O-ring 814. As described above, in some embodiments, O-ring fluid connection 812 may be fluidly connected to a flow path defined by inlet 802 and outlet 804 and may be configured to direct the flow of fluid (e.g., cell suspension and / or buffer fluid) into and / or out of constriction component 806.

[0111] 8A-8D shown herein discuss constriction components arranged on different outward-facing surfaces of constriction cartridge 800, different arrangements for cartridges containing two or more constriction components may be used in some embodiments. For example, in some embodiments, the constriction cartridge may include three or more slots for inserting constriction components such that the constriction components are arranged in a layered, stacked arrangement (e.g., similar to the arrangement of constriction component 706 in constriction cartridge 700) and the cartridge may be configured to direct flow in parallel into and through each of the stacked components.

[0112] 9A-9D illustrate a sensor assembly 900 of tabletop system 100 for delivering a payload to a cell, according to some embodiments. In some embodiments, sensor assembly 900 may be configured to transmit electronic signals to one or more electronic components of system 100, as discussed above with respect to FIG. 1. In some embodiments, sensor assembly 900 may include one or more sensors configured to measure one or more properties of a component of system 100 or of a substance contained within system 100 and transmit data regarding the measurements made to the electronic components of system 100. In some embodiments, sensor assembly 900 may contain one or more sensors configured to measure one or more properties. For example, the sensor assembly may comprise a temperature sensor, a presence sensor, a flow sensor, and / or a pressure sensor.

[0113] That is, the sensor assembly 900, in some embodiments, may be configured to make one or more measurements and send signals to the electronic components of the system 100 regarding the state of the cell suspension as it passes through the system 100, such that the system may monitor the state of the cell suspension (and, optionally, control the state of the cell suspension accordingly). In some embodiments, the sensor assembly 900 may be configured to measure and / or monitor the temperature of the cell suspension (or the temperature of a container holding the cell suspension), such as while the cell suspension is in a preparation vessel and being heated or cooled by the system. In some embodiments, the sensor assembly 900 may be configured to measure and / or monitor the pressure being applied to the cell suspension, such as the pressure of a gas inside a preparation vessel as the cell suspension accumulates inside the preparation vessel and / or flows through a flow path downstream of the preparation vessel. In some embodiments, the sensor assembly 900 may be configured to monitor the flow path of the cell suspension and determine whether the cell suspension is present in the flow path, such as by using an optical presence sensor to determine whether the cell suspension is flowing through a tube to determine whether the cell suspension has finished flowing through a constriction cartridge.

[0114] 9A-9D, in the example, FIG. 9A illustrates a partially assembled sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments. FIG. 9B illustrates a detailed view of the sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments. FIG. 9C illustrates a detailed view of the sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments. And FIG. 9D illustrates the sensor assembly of a tabletop system for delivering a payload to a cell, according to some embodiments.

[0115] The sensor assembly 900 may include a multi-pin sensor wire connector 904 at one end, with one or more wires 901 extending longitudinally along the length of the assembly and a temperature sensor connector 906 disposed at the opposite end of the wires 901. In some embodiments, the multi-pin wire connector 904 is an electronic connector configured to allow connection of a sensor with one or more electronic interfaces such that electronic signals, such as signals representing data measured or detected by one or more sensors, can be transmitted and received through the connector 904. In some embodiments, the multi-pin wire connector 904 may be configured such that one or more pins of the connector can correspond to different sensors and may be configured to transmit and receive different types of data. In some embodiments, instead of or in addition to a wire connector, an alternative type of electronic connector configured to transmit and receive data to and from other portions of the system 100 may be used as part of the sensor assembly 900. In some embodiments, the sensor assembly 900 may be configured to be disposable and capable of being used in a sterile environment, such that the connector 904 may be configured to be easily attached to and detached from the electronic interface by hand and / or without the use of tools.

[0116] Sensor assembly 900 may further include a stenosis cartridge seat 902, which may be a component mounted on wire 901 and configured to retain the stenosis cartridge, such as by removably clipping onto the stenosis cartridge. In some embodiments, cartridge seat 902 may be formed to allow attachment of one or more types of stenosis cartridges, such as stenosis cartridge 700 or stenosis cartridge 800 as discussed above. In some embodiments, sensor assembly 900 may be configured to be disposable and capable of being used in a sterile environment, such that cartridge seat 902 may be configured to allow easy attachment and detachment from the stenosis cartridge by hand and / or without the use of tools. In the example of FIGS. 9A-9D , stenosis cartridge seat 902 includes a clip 910 that may hold the stenosis cartridge in place under tension. In alternative embodiments, different attachment means may be used.

[0117] In some embodiments, the constriction cartridge seat 902 may be configured to attach to the wire 901 such that when the constriction cartridge is attached to the seat 902, the wire 901 extends in the same or similar linear direction as a linear direction defined by a flow path leading to and from the constriction cartridge inlet and outlet. In this manner, the sensor assembly 900 may be configured such that the wire 901 and the tube leading to / from the constriction cartridge may extend alongside one another and may be able to pass through one or more of the same openings as one another, such as by passing through an outlet of the preparation vessel housing, such as outlet 514 as discussed above with reference to the preparation vessel housing 110.

[0118] Sensor assembly 900 may include a temperature sensor connector 906, which may be any component configured to physically and / or electronically connect the temperature sensor to other components of sensor assembly 900. In the example of Figures 9A-9D, temperature sensor connector 906 may be an electrical connector disposed at the opposite end of wire 901 from connector 904 and configured to removably physically and electronically connect to a temperature sensor (e.g., a thermistor or temperature probe). As shown in Figure 9D, temperature sensor 914 may be physically and electronically attached to temperature sensor connector 906 such that temperature sensor 914 may transmit a signal related to temperature data measured by temperature sensor 914 via connector 906 (as well as through wire 901 and connector 904).

[0119] In some embodiments, the temperature sensor 914 may be any device configured to measure one or more temperatures and generate and / or transmit a signal related to the one or more temperatures measured by the device. In some embodiments, the temperature sensor 914 may be any suitable type of temperature probe or thermistor. In some embodiments, the temperature sensor 914 may be an adhesive temperature probe configured to be adhered to a wall of the preparation vessel or preparation vessel housing to measure a temperature associated with a fluid (e.g., a cell suspension) contained within the vessel.

[0120] In some embodiments, by being positioned at the end of wire 901 opposite connector 904, temperature sensor 914 can be positioned such that it will be in proximity to the preparation vessel or preparation vessel housing when sensor assembly 900 is connected to system 100. For example, when the constriction cartridge is connected to seat 902, such that fluid may flow through the constriction cartridge from right to left with respect to FIG. 9A , temperature sensor 914 can be positioned in proximity to the preparation vessel or preparation vessel housing, from which fluid flows toward and through the constriction cartridge. In this manner, temperature sensor 914 may be positioned in physical contact with the preparation vessel or its housing to measure the temperature associated with the fluid in the preparation vessel while sensor assembly 900 and associated chip cartridge are fully assembled into system 100.

[0121] The sensor assembly 900 may further include a flow sensor connector 908, which may be any component configured to physically and / or electronically connect the flow sensor to other components of the sensor assembly 900. In the example of FIGS. 9A-9D , the flow sensor connector 908 may be an electrical connector disposed at one end of the cartridge seat 902 and configured to physically and electronically connect to a flow sensor (e.g., an optical presence sensor, a capacitive sensor, a weight sensor, or other sensor configured to determine whether flow is occurring). As shown in FIGS. 9C and 9D , the flow sensor 912 may be physically and electronically attached to the flow sensor connector 908 such that the flow sensor 912 may transmit signals related to data measured by the flow sensor 912 via the connector 908 (and through the wire 901 and the connector 904).

[0122] In some embodiments, the flow sensor 912 may be any sensor configured to measure or otherwise determine whether flow is occurring within a particular portion of a flow path, either by making an analog determination as to whether flow is occurring or by measuring the flow rate. In some embodiments, the flow sensor 912 may be an optical sensor configured to use the presence, absence, and / or change of fluid in the path of a light beam to determine whether fluid is flowing through the path. For example, in some embodiments, the flow sensor 912 may be a foam sensor configured to determine whether flow is occurring through a translucent or transparent tube extending through the path of the foam sensor's light beam. In some embodiments, the flow sensor 912 may be a capacitive sensor configured to sense the presence of liquid, a weight sensor configured to sense the weight of liquid, or a physical sensor configured to be installed within the flow path to measure the velocity and / or flow rate of the fluid.

[0123] In some embodiments, by being disposed on or near cartridge seat 902, flow sensor 912 may be positioned so that it will be in the vicinity of the constriction cartridge when sensor assembly 900 is connected to system 100. For example, if the constriction cartridge is connected to seat 902 such that fluid may flow through the constriction cartridge from right to left with respect to FIG. 9A , flow sensor 912 may be positioned near the inlet of the constriction cartridge such that the flow sensor may measure fluid flow through the tube or pipe as it approaches (or alternatively, as it flows away from) the constriction cartridge. For example, if the flow sensor is an optical bubble sensor, the tubing leading to the inlet of the constriction cartridge may be configured to be seated within the cavity of the bubble sensor such that the tubing is in the optical path from which the sensor derives measurements. In this way, flow sensor 912 performs measurements to determine whether fluid (e.g., a cell suspension) is flowing toward, away from, and / or through the constriction cartridge while sensor assembly 900 and chip cartridge are fully assembled into system 100.

[0124] FIG. 10 illustrates a schematic diagram of a tabletop system 1000 for delivering a payload to cells, according to some embodiments. In some embodiments, system 1000 may share some or all characteristics in common with system 100 and / or any other systems for delivering a payload to cells discussed herein. Rather than depicting the physical shapes of the various components of a system for delivering a payload to cells, FIG. 10 primarily depicts diagrammatically the flow paths and associated components for fluids (e.g., cell suspension, buffer fluid) to travel through the system and for pressurized gas to travel through the system. That is, FIG. 10 depicts various components through which cell suspension and / or buffer fluid may flow while being processed by the system, and various components through which gas (e.g., pressurized gas) may flow as it is being passed through the system. FIG. 10 shows components fluidly connected to one another via arrows and lines connecting the depictions of the components. Unless otherwise stated, any suitable tubing or plumbing may be used to fluidly connect the various components, such as flexible plastic tubing, rigid plastic tubing, PVC tubing, metal tubing, or the like.

[0125] The system 1000 may include a cell suspension input bag 1002 that may share some or all characteristics in common with the cell suspension input bag 106 discussed above with reference to Figure 1. In some embodiments, the flow path of the liquid flowing through the system 1000 may originate (or part of) from the cell suspension input bag 1002.

[0126] System 1000 may include a buffer input bag 1004 that may share some or all characteristics in common with buffer input bag 108 discussed above with reference to Figure 1. In some embodiments, the flow path of liquid flowing through system 1000 may originate (or part of) from buffer input bag 1004.

[0127] System 1000 may include a tubing clamp 1006 that may be used to prevent liquid from flowing through the tubing extending from cell suspension input bag 1002 or buffer input bag 1004. In some embodiments, other mechanisms for preventing flow from the bags, such as valves, caps, or the like, may be used as an alternative to or in addition to clamp 1002. In some embodiments, clamp 1006 may be configured to be capable of being automatically actuated by an electronic control system such that manual actuation by a user is not required.

[0128] System 1000 may include fitting 1008, which may be any connector, configured to fluidly connect tubing or tubing extending from bags 1002 and 1004 to additional components of system 1000 defining further downstream portions of the flow path. In some embodiments, fitting 1008 may be a connector mechanism configured to provide a sterile connection (e.g., a connector mechanism configured to ensure that the interior of the flow path is not contaminated from direct handling as the connection is secured), such as an ASEPTIQUIK sterile fitting. In some embodiments, one or more connection mechanisms other than fittings may be used in addition to or instead of fitting 1008, such as a tube weld connection, a sterile assembly connection, etc.

[0129] System 1000 may include a Y-joint 1010, which may be any diverter or junction configured to converge the flow paths from bag 1002 and bag 1004 into a single flow path.

[0130] System 1000 may include a fitting 1012, which may be any suitable tubing or piping fitting or connector for connecting the tubing or piping of the flow path of system 1000 to valve 1014 so that fluid may flow into valve 1014. In some embodiments, fitting 1012 may be a threaded fitting and / or a luer fitting.

[0131] System 1000 may include valve 1014, which may be any valve configured to control the flow of fluid from fitting 1012 to and into vessel 1016. In some embodiments, valve 1014 may be a syringe valve, a manual valve, an electronic valve, and / or a solenoid valve. In some embodiments, valve 1014 may be configured to be capable of being automatically actuated by an electronic control system such that manual actuation by a user is not required.

[0132] System 1000 may include a vessel 1016 that may share any one or more characteristics in common with preparation vessel 600 discussed above with respect to Figures 6A and 6B.

[0133] System 1000 may include connector 1018, which may share any one or more characteristics in common with connector 1012. Connector 1018 may be configured to connect tubing or plumbing of a flow path of system 1000 to vessel 1016 so that fluid may flow out of vessel 1016.

[0134] System 1000 may include constriction cartridge 1020, which may share any one or more characteristics in common with constriction cartridge 700 and / or constriction cartridge 800 described above with respect to Figures 7A-7C and 8A-8D. Although system 1000 is shown with only one constriction cartridge 1020, some embodiments of system 1000 (or other systems described herein) may include multiple constriction cartridges, which may be arranged in parallel and / or series with one another. In some embodiments, multiple constriction cartridges within the same system may be associated with the same or separate sensor assemblies, the same or separate preparation vessels, and / or the same or separate set input / output containers.

[0135] System 1000 may include connector 1022, which may share any one or more characteristics in common with connector 1012. Connector 1022 may be configured to connect tubing or plumbing of the flow path of system 1000 to valve 1024 such that fluid may flow from constriction cartridge 1020 toward and into valve 1024.

[0136] System 1000 may include valve 1024, which may share some or all characteristics in common with valve 1014. Valve 1024 may be configured to control the flow of fluid from connector 1022 to and into connector 1026, or more broadly, the flow of fluid from constriction cartridge 1020 to and into output bags 1032 and 1039.

[0137] System 1000 may include connector 1026, which may share any one or more characteristics in common with connector 1012. Connector 1026 may be configured to connect tubing or plumbing of the flow path of system 1000 to valve 1024 such that fluid may flow out of syringe 1024 toward output bags 1032 and 1039.

[0138] The system 1000 may include a Y-joint 1028, which may be any diverter or junction configured to split the flow path from the connector 1026 into two separate flow paths, one leading toward the cell suspension output bag 1032 and the other leading toward the buffer output bag 1039.

[0139] System 1000 may include clamp 1030, which may share any one or more characteristics in common with clamp 1006. In some embodiments, clamp 1030 may be used to prevent liquid from flowing through the tubing leading to cell suspension output bag 1032 or buffer output bag 1039. In some embodiments, other mechanisms for preventing flow into the bags, such as valves, caps, or the like, may be used as an alternative to or in addition to clamp 1030. In some embodiments, clamp 1030 may be configured to be capable of being automatically actuated by an electronic control system, such that manual actuation by a user is not required.

[0140] The system 1000 may include a cell suspension output bag 1032 that may share any one or more characteristics in common with the cell suspension output bag 112 as described above with respect to Figure 1. In some embodiments, the flow path of the liquid flowing through the system 1000 may terminate (or be partially terminated) in the cell suspension input bag 1002.

[0141] System 1000 may include a buffer output bag 1034 that may share any one or more characteristics in common with buffer output bag 114 as described above with respect to Figure 1. In some embodiments, the flow path of liquid flowing through system 1000 may terminate (or be partially terminated) in buffer output bag 1034.

[0142] System 1000 may include connector 1036, which may share any one or more characteristics in common with connector 1012. Connectors 1036 may each connect to a gas outlet line extending from one of output bags 1032 and 1039, allowing gas to be forced out of the output bag before, during, or after the process of flowing the cell suspension and / or buffer fluid through system 1000 and into the output bag. For example, because fluids may be forced through system 1000 under the force of pressurized air, the output bag, which may form the termination point of the flow path for the cell suspension and / or buffer fluid through system 1000, may need to have a gas outlet to prevent the bag from bursting under pressure.

[0143] System 1000 may include filter 1038, which may be a gas filter, connected to one of connectors 1036 so that air or other gas may flow through one of the output bags, through one of filters 1038, and into the environment external to system 1000. In some embodiments, filter 1038 may ensure that gas exhausted into the environment external to system 1000 is suitable for a sterile environment. In some embodiments, one or both of filters 1038 may share one or more characteristics in common with filter 404 as discussed above with respect to FIG. 4. For example, filter 1038 may be a HEPA filter suitable for use in filtering exhaust air to be exhausted into a sterile laboratory environment.

[0144] System 1000 may include a filter subassembly 1042, which may be configured to be pre-sterilized or capable of being sterilized (e.g., by autoclaving or by ethylene oxide sterilization). In some embodiments, filter subassembly 1042 may be configured to receive a gas, such as pressurized sterile gas (e.g., pressurized sterile nitrogen), from an inlet, direct a portion of the gas into container 1016, and direct a portion of the gas to an outlet. As shown in FIG. 10 , filter subassembly 1042 may receive gas from gas inlet 1044, direct some or all of the received gas into container 1016, and direct some or all of the received gas to outlet 1058. In some embodiments, filter subassembly 1042 may be used to direct pressurized sterile gas into container 1016 to apply pressure to a fluid (e.g., a cell suspension) within container 1016 to urge the fluid to flow under pressure out of container 1016 through constriction cartridge 1020. 10, system 1000 may, in some embodiments, include multiple filters in series, as redundancy may improve reliability and safety. In some embodiments, filter subassembly 1042 may be located outside the housing of system 1000 so that filter subassembly 1042 may be easily replaced.

[0145] System 1000 may include a gas inlet 1044, which may comprise any suitable flexible or rigid inlet configured to receive a flow of gas, such as pressurized sterilizing gas. In some embodiments, gas inlet 1044 may be configured to be fluidly connectable to flexible tubing for gas and / or rigid tubing for gas. In some embodiments, gas inlet 1044 may be configured to be fluidly connectable to the tubing by a clamp, threads, a luer connector, or any other suitable connection mechanism.

[0146] System 1000 may include gas outlet 1058, which may be any suitable flexible or rigid outlet configured to exhaust a flow of gas, such as pressurized sterilizing gas. In some embodiments, gas outlet 1058 may be configured to be fluidly connectable to flexible tubing for gas and / or rigid tubing for gas. In some embodiments, gas outlet 1058 may be configured to be fluidly connectable to the tubing by a clamp, threads, a luer connector, or any other suitable connection mechanism.

[0147] System 1000 may further include connectors 1046, 1048, 1050, 1052, 1054, and 1056, any one or more of which may share any one or more characteristics in common with connector 1012. Connector 1012 may be configured to connect to tubing or piping or other flow path elements for the flow path of liquids, while connectors 1046, 1048, 1050, 1052, 1054, and 1056 may be configured to connect to tubing or piping or other flow path elements for the flow path of gases. As shown in FIG. 10 , connectors 1046, 1048, 1050, 1052, 1054, and 1056 may be used to connect tubing or piping elements between gas inlet 1044, filter subassembly 1042, vessel 1016, and gas outlet 1058. In this manner, connectors 1046, 1048, 1050, 1052, 1054, and 1056 may allow gas to flow from the inlet, through the filter subassembly, and to the outlet reservoir.

[0148] System 1000 may include a sensor assembly 1060 that may share any one or more characteristics in common with sensor assembly 900 discussed above with reference to Figures 9A-9D. In some embodiments, sensor assembly 1060 may include one or more sensors not depicted in Figure 10, such as a temperature sensor.

[0149] System 1000 may include a flow sensor 1062, which in some embodiments may be part of a sensor assembly 1060. In some embodiments, flow sensor 1062 may share one or more characteristics in common with flow sensor 912 discussed above with reference to FIGS. 9C and 9D.

[0150] In some embodiments, one or more components shown in FIG. 10 may together form all or part of a disposable assembly. For example, the components may be configured for one-time use such that they can be used to perform a payload delivery process once and then discarded. That is, a cell suspension may be flowed through the flow paths of system 1000 once, and then some or all of the elements of system 1000 may be replaced before another payload delivery process is performed. In some embodiments, the components of the disposable assembly may be constructed from materials suitable for gamma sterilization to be suitable for use in a sterile environment. In some alternative embodiments, the components of the disposable assembly may be constructed from materials suitable for sterilization by other methods, such as autoclaving or ethylene oxide sterilization. In some embodiments, the components of the disposable assembly may be packaged and / or shipped together, such as packaged and / or shipped in a sealed sterile container. In some embodiments, the components of the disposable assembly may be configured to be capable of being attached to other components of a system for intracellular payload delivery in a manner suitable for implementation in a sterile environment, such as by hand, without the use of tools, and / or by using a sterile connector mechanism. In the example of Figure 10, the disposable assembly 1066 may include all of the components shown inside the dotted box indicated by reference numeral 1066.

[0151] 11A and 11B illustrate schematic diagrams of a system 1100 for supplying pressurized gas for use in delivering a payload to cells, according to some embodiments. In some embodiments, system 1100 may be configured to supply pressurized gas to a system such as system 1000 as discussed above with reference to FIG. 10. In some embodiments, system 1100 may form part of, or share some or all characteristics in common with, pressure control module 116 discussed above with respect to FIGS. 1 and 2. As shown in FIGS. 11A and 11B, system 1100 may include flow paths for gas, such as pressurized sterilizing gas (e.g., pressurized sterilizing nitrogen), to flow from an inlet source to and into a system having a vessel containing a fluid (e.g., a cell suspension), and to and out of an outlet into an environment external to system 1100 and / or outside an associated system, such as system 1000. In some embodiments, system 1100 may be used to direct pressurized sterilizing gas into container 1016 to apply pressure to a fluid (e.g., a cell suspension) within container 1016 to force the fluid to flow under pressure out of container 1016 through constriction cartridge 1020.

[0152] 11A and 11B show components fluidly connected to one another via arrows and lines connecting the depictions of the components. Unless otherwise stated, any suitable tubing or plumbing, such as flexible plastic tubing, rigid plastic tubing, PVC tubing, metal tubing, or the like, may be used to fluidly connect the various components.

[0153] 11A, system 1100 may include inlet 1101, which may be any inlet configured to be fluidly connected to a source of gas, such as pressurized sterilizing gas (e.g., pressurized sterilizing nitrogen). In some embodiments, inlet 1101 may comprise any suitable flexible or rigid inlet and / or connector configured to receive a flow of gas, such as pressurized sterilizing gas. In some embodiments, inlet 1101 may be configured to be fluidly connected to flexible tubing for gas and / or rigid tubing for gas, may be configured to be fluidly connected to a pressurized gas canister, and / or may be configured to draw gas from the environment (e.g., from air). In some embodiments, inlet 1101 may be configured to be fluidly connectable to tubing by a clamp, threads, a luer connector, or any other suitable connection mechanism to direct the flow of gas toward regulator 1102.

[0154] System 1100 may include a regulator 1102 that may be fluidly connected to inlet 1101 and configured to receive a flow of gas therefrom. In some embodiments, regulator 1102 may be configured to receive the flow of gas and output the gas at a predetermined pressure. In some embodiments, the pressure of the gas output by regulator 1101 may be controlled by a user or automatically controlled by the system. In some embodiments, the pressure of the gas output by regulator 1101 may be variable by electronic control such that no manual intervention is required.

[0155] System 1100 may include filter 1104, which may be fluidly connected to regulator 1102 and configured to receive a flow of gas therefrom. In some embodiments, filter 1104 may be any filter configured to allow gas output by regulator 1102 to pass therethrough before flowing along the flow path to regulator 1106. For example, filter 1104 may be a HEPA filter, such that system 1100 and associated systems may be suitable for use in a sterile environment.

[0156] System 1100 may include a regulator 1106 fluidly connected to filter 1104 and configured to receive a gas flow therefrom. In some embodiments, regulator 1106 may be configured to receive a gas flow at one pressure and output a gas flow at another, lower, user- or system-selectable pressure. In some embodiments, regulator 1106 may be an electro-pneumatic regulator and may share some or all properties in common with regulator 204 as described above with respect to FIG. 2. In some embodiments, regulator 1106 may include a silencer to reduce vibration and noise. The use of a silencer to reduce vibration and noise may make system 1100 suitable for use in a more sterile environment in that it may minimize agitation of particles in the environment. In some embodiments, regulator 1106 may be configured for more precise pressure control over a narrower range than regulator 1102, which may be configured to more roughly control pressure over a wider range.

[0157] System 1100 may include a valve 1108 that may be fluidly connected to regulator 1106 and configured to receive the flow of gas therefrom. Valve 1108 may be configured to block or allow the flow of gas toward diverter 1110. In some embodiments, valve 1108 may be manually and / or electronically actuable, and valve 1108 may be actuated by a user or by an electronic system pursuant to operation of the system and without user intervention.

[0158] The system 1100 may include a diverter 1110 that may be fluidly connected to the valve 1108 and configured to receive a flow of gas therefrom. In some embodiments, the diverter 1110 may be any piping or tubing element configured to split a single flow path into one or more flow paths. In the example shown in FIG. 11A , the diverter 1110 splits the flow path extending from the valve 1108 into two flow paths, one flowing toward a pressure sensor 1112 and the other flowing toward a fitting 1114 in FIG. 11B . In some embodiments, the diverter 1110 may include one or more valves that can be opened or closed to selectively split the flow of gas along one path or another. In some embodiments, the diverter 1110 may be configured such that both downstream flow paths are always open and gas is always directed into both flow paths.

[0159] System 1100 may include a pressure sensor 1112 that may be fluidly connected to diverter 1110 and configured to receive a flow of gas therefrom. In some embodiments, pressure sensor 1112 may be configured to measure the pressure of the gas in a flow path of system 1100 between valve 1108 and fitting 1114. In some embodiments, pressure sensor 1112 may be configured to generate data representative of the obtained pressure measurements and transmit electronic signals representative of the data to one or more other components of system 1100, system 1000, or an associated system. For example, pressure sensor 1112 may transmit a signal related to the measured pressure to regulator 1106 so that regulator 1106 may make adjustments to the pressure of the gas output by regulator 1106 as required. In some embodiments, for example, the gas pressure may drop as the cell suspension is forced out of the preparation vessel and into and through the constriction cartridge, and pressure sensor 1112 may sense this decrease in pressure and send a signal to a processor in the system (e.g., regulator 1106 and / or another component of the pressure control module), causing the system to adjust one or more valves to cause the pressure to remain relatively constant as the cell suspension is forced out of the vessel.

[0160] 11B, system 1100 may include a fitting 1114 that may be fluidly connected to diverter 1110 and configured to receive the flow of gas therefrom and direct it toward system 1000. In some embodiments, fitting 1114 may be any connector configured to fluidly connect with flexible tubing for gas and / or rigid tubing for gas. In some embodiments, fitting 1114 may be configured to be fluidly connectable to rigid tubing on the upstream side and flexible tubing on the downstream side. In some embodiments, fitting 1114 may be configured to be fluidly connectable to tubing with a clamp, threads, a luer connector, or any other suitable connection mechanism to direct the flow of gas toward system 1000.

[0161] 11B, fitting 1114 may connect a component of system 1100 to one or more components of system 1000. In particular, fitting 1114 may be configured to be fluidly connected (and / or connectable) to a flow path leading to inlet 1044 of system 1000. In some embodiments, gas delivered from system 1100 to system 1000 via fitting 1114 and inlet 1044 may be used to apply pressure to a fluid inside vessel 1016, as described above with reference to FIG.

[0162] System 1100 may include fitting 1120, which may share any one or more characteristics in common with fitting 1114. Fitting 1120 may be configured to be fluidly connected (or connectable) to a flow path leading from outlet 1058 of system 1000. In some embodiments, gas output by outlet 1058 in system 1000 as described with reference to FIG. 10 may flow through fitting 1120 into the flow path depicted in FIGS. 11A and 11B.

[0163] 11A, system 1100 may include a silencer 1122. In some embodiments, silencer 1122 may be configured to allow gas to flow through it before being exhausted into the environment (e.g., into the open air) external to system 1100. In some embodiments, silencer 1122 may make system 1100 suitable for use in a more sterile environment in that it may reduce vibration and noise and minimize agitation of particles in the environment. Intracellular payload delivery and cell processing methods and techniques

[0164] 12-13 illustrate various methods that may be implemented, in whole or in part, by one or more of the systems described herein.

[0165] 12 illustrates a method 1200 for processing cells, including intracellular payload delivery, according to some embodiments. As shown in FIG. 12, method 1200, in some embodiments, can be performed over the course of 24 hours or less. In some alternative embodiments, method 1200 may be performed in 72 hours or less, 36 hours or less, 18 hours or less, or 12 hours or less. In some embodiments, performing method 1200 in less than one of the time frames shown may improve throughput and efficiency for cell processing and intracellular payload delivery techniques. In some embodiments, the time frames discussed herein may be facilitated by high-throughput intracellular payload delivery techniques enabled by the systems and methods described elsewhere herein.

[0166] In step 1202, in some embodiments, a supply of cells is received. In some embodiments, the supply of cells may comprise various blood cells, including mononuclear cells, lymphocytes, platelets, plasma, and red blood cells. In some embodiments, the supply of cells may comprise an enriched leukapheresis product, such as LEUKOPAK or a similar product, which may be delivered overnight at room temperature.

[0167] In some embodiments, the supply of cells may be processed for lymphocyte enrichment in step 1204. In some embodiments, lymphocyte enrichment may be performed by an ELUTR cell separation system or a similar system.

[0168] In some embodiments, the cells may be processed for washing and / or buffer exchange in step 1206. In some embodiments, cell washing and / or buffer exchange may be performed by a LOVO automated cell processing system or a similar system.

[0169] In step 1208, in some embodiments, the cells may be incubated. In some embodiments, the incubation may be a bead incubation. In some embodiments, the bead incubation may be performed using polymer resin beads. In some embodiments, the bead incubation may be performed using a SEPAX system and / or SEPAX polymer resin beads, or similar systems and / or materials.

[0170] In step 1210, in some embodiments, target cells may be isolated from the cells. In some embodiments, the target cells to be isolated may comprise NK cells, T cells, B cells, and / or other cell types. In some embodiments, the target cells may be isolated using a positive selection process (e.g., tagging the target cells) or a negative selection process (e.g., tagging cells other than the target cells). In some embodiments, the cells may be separated based on their relative density. In some embodiments, isolating the target cells may include isolating a single type of cell, while in some embodiments, it may include isolating multiple different types of cells, all of which are target cells. In some embodiments, the target cells may be isolated by a CLINIMACS automated cell separation system or a similar system.

[0171] In step 1212, in some embodiments, the target cells may be processed for post-isolation incubation preparation. In some embodiments, the incubation preparation process may prepare the target cells for CO incubation. In some embodiments, the post-isolation incubation preparation process may be performed by a LOVO automated cell processing system or a similar system.

[0172] In some embodiments, the target cells may be incubated in step 1214. In some embodiments, the incubation may be a CO2 incubation at about 37°C.

[0173] In step 1216, in some embodiments, the target cells may be prepared for a payload delivery process, which may include creating a perturbation in the membrane of the target cells to allow the payload material to enter the target cells. In some embodiments, the payload delivery process preparation may include removing the original buffer and / or suspending the target cells in a fluid (e.g., a delivery buffer) to create a cell suspension. While in some embodiments the cell suspension may include a payload to be delivered to the cells, in some embodiments the payload may not be contained within the cell suspension (and the payload may instead be contacted with the cells after the cell suspension has been passed through all or part of the system, while the cell suspension is inside the preparation vessel, before all or part of the cell suspension is passed through the disposable assembly, before all or part of the cell suspension is passed through the constriction cartridge, after all or part of the cell suspension has passed through the disposable assembly, after all or part of the cell suspension has passed through the constriction cartridge, before some or all of the cell membranes are perturbed by being passed through one or more constrictions, after some or all of the cell membranes are perturbed by being passed through one or more constrictions, inside the preparation vessel, inside the constriction cartridge, inside the output bag, and / or inside any one or more other portions of the flow path of the system).

[0174] In step 1218, in some embodiments, the target cells may be processed by an intracellular payload delivery system to deliver the payload to the target cells. In some embodiments, the intracellular payload delivery system may cause perturbations in the membrane of the target cells, thereby passing the target cells through one or more constrictions to allow entry of the payload into the target cells. In some embodiments, the intracellular payload delivery system may share any one or more properties in common with systems 100, 1000, and / or 1100 as described above. Intracellular payload delivery techniques, particularly as may be performed by a tabletop system for payload delivery, will be discussed in more detail below with respect to FIG. 13.

[0175] In some embodiments in which intracellular payload delivery is performed by an intracellular payload delivery system, such as systems 100, 1000, and / or 1100, the system may be configured to be attached to one or more other cell processing systems or devices described herein and / or used anywhere in method 1200. For example, in some embodiments, the cell suspension may be configured to flow directly from a separate device into one or more components of the intracellular payload delivery system, such as into an input bag, into a preparation vessel, and / or into a constriction cartridge (whether or not initially held within the preparation vessel).

[0176] In step 1220, in some embodiments, the target cells with the delivered payload may be processed for post-payload delivery incubation preparation. In some embodiments, the incubation preparation process may prepare the target cells for CO incubation. In some embodiments, the post-payload delivery incubation preparation process may be performed by a LOVO automated cell processing system or a similar system.

[0177] In some embodiments, the target cells may be incubated in step 1222. In some embodiments, the incubation may be a CO2 incubation.

[0178] In some embodiments, the target cells may be prepared for cryogenic storage in step 1224. In some embodiments, the cryogenic storage preparation may be performed by using a SEPAX system or a similar system.

[0179] In some embodiments, the target cells may be placed in a cryogenic vial in step 1226. In some embodiments, the cryogenic vial may be any sterile vial suitable for cryogenic preservation and storage.

[0180] In some embodiments, the cryogenic vial containing the target cells with the payload to be delivered may be frozen in step 1228. In some embodiments, the freezing process may be performed at a controlled rate by a controlled rate freezer system, which may prevent damage to the cells by preventing them from freezing too rapidly.

[0181] Finally, in step 1230, in some embodiments, the frozen cryogenic vials may be stored in a liquid nitrogen storage system.

[0182] FIG. 13 illustrates a method 1300 for intracellular payload delivery, according to some embodiments. In some embodiments, all or a portion of method 1300 may be implemented as step 1218 of method 1200 as described above with reference to FIG. 12. In some embodiments, method 1300 may be used to process cells with an intracellular payload delivery system to deliver a payload to the cell. In some embodiments, the intracellular payload delivery system may cause a perturbation in the cell's membrane, thereby passing the cell through one or more constrictions to allow entry of the payload into the cell. In some embodiments, the intracellular payload delivery system may share any one or more properties in common with systems 100, 1000, and / or 1100 as described above. Below, method 1300 will be described primarily with reference to the components of system 100 as described above in FIGS. 1-9D.

[0183] At block 1302, in some embodiments, cells in a cell suspension may be provided to the intracellular payload delivery system. In some embodiments, the cells in cell suspension may be provided in the form of a cell suspension fluid provided in a cell suspension input bag, such as bag 106 of system 100. Providing the cells in cell suspension may include suspending bag 106 from hook 104 and attaching tubing from bag 106 to an inlet of system 100, such as an inlet provided on one of preparation vessel housing 110 and / or fittings 1008 in system 1000.

[0184] As will be discussed in more detail below, in some embodiments, the cells may be provided as part of a cell suspension, which may contain a payload for delivery to the cells. In some other embodiments, the cells may be provided as part of a cell suspension that does not contain a payload for delivery to the cells, and the payload may instead be contacted with the cells and / or cell suspension at a later time.

[0185] At block 1304, in some embodiments, a disposable assembly comprising a preparation vessel may be attached to the intracellular payload delivery system. In some embodiments, the disposable assembly may comprise a preparation vessel, such as preparation vessel 600, a constriction cartridge, such as constriction cartridge 700 and / or constriction cartridge 800, and / or a sensor assembly, such as sensor assembly 900. In some embodiments, the disposable assembly may comprise any one or more of the components included within disposable assembly 1066 of system 1000. In some embodiments, attaching the disposable assembly to the system may include physically, fluidly, and / or electronically / communicatively connecting one or more components of the disposable assembly to the system. For example, the preparation container of the disposable assembly may be physically attached to the system by being placed inside a receptacle or housing, one or more tubes or pipes of the disposable assembly may be fluidly connected to the system (e.g., by a Luer connector) so that liquids and / or gases may flow through the tubes or pipes, and one or more electronic connectors of the sensor assembly may be electronically communicatively coupled to the system so that signals and data from the sensors of the sensor assembly may be transmitted to the electronic components of the system.

[0186] At block 1306, in some embodiments, attaching the disposable assembly may include inserting the preparation vessel of the disposable assembly into the preparation vessel housing. In an example of system 100, the preparation vessel 600 may be inserted into the housing 110, and the door of the housing 110 may be closed and latched. As discussed above, closing and latching the door of the housing 110 may urge the preparation vessel 600 into contact with the interior wall of the housing 110 so that optimized heat transfer between the vessel 600 and the housing 110 may be achieved. In some embodiments, attaching the preparation vessel may further include fluidly connecting the preparation vessel to one or more connectors, such as inlet 604, to establish a flow path for liquid and / or gas to enter the preparation vessel.

[0187] In block 1308, in some embodiments, attaching the disposable assembly may include attaching one or more sensors of the disposable assembly to the system or to one of the system components. For example, attaching the disposable assembly may include attaching tubing leading from an outlet of the preparation vessel to a flow sensor. In an example of the sensor assembly 900, the tubing may be attached to a slot in the flow sensor 912 so that the sensor can monitor the flow through the tubing. Alternatively, or in addition, attaching the disposable assembly may include attaching a temperature sensor to the preparation vessel. In an example of the sensor assembly 900, the temperature sensor 914 may be adhered to an exterior surface of the preparation vessel so that it can monitor a temperature associated with the contents of the preparation vessel.

[0188] In addition to providing a physical connection, attaching the disposable assembly to the system may further include establishing an electronic communication connection so that one or more sensors of the disposable assembly may transmit data to the electronic and control components of the system. In an embodiment of the sensor assembly 900, the connector 904 may plug into an electronic data interface of the system 100 to transmit data from the flow sensor and / or the temperature sensor to the system 100.

[0189] In block 1310, in some embodiments, attaching the disposable assembly may include attaching one or more components of the disposable assembly by hand and / or without the use of tools. For example, the components of the disposable assembly may be configured to be able to be attached and removed using threads, luer connectors, latches, plugs, and other connection mechanisms configured to be operated by hand. In this manner, the disposable assembly may be able to be used without tools in a sterile environment, which may increase availability due to the space occupied by tools and the fact that tools may become contaminated and unable to be used in a sterile environment.

[0190] At block 1312, in some embodiments, the system may perform an integrity test. In some embodiments, the integrity test is an integrity test to test the integrity of one or more components of the most recently installed disposable assembly. For example, an integrity test may be performed to determine whether all components of the disposable assembly are physically, fluidly, and electronically connected in a predetermined manner, such that the system can use one or more sensors to determine whether the components are installed in a predetermined manner and provide an output to the user indicating so.

[0191] Additionally, an integrity test may be performed to determine whether one or more components of the disposable assembly are capable of maintaining an internal gas pressure at the system's operating pressure. As discussed above, the system's operating pressure may be the pressure used to force fluid through the constriction cartridge. In some embodiments, testing the pressurized integrity via an integrity test may include forcing pressurized gas into the preparation vessel and / or other portions of the system's flow path and monitoring the pressure using one or more pressure sensors in the system. Once the system is pressurized to the operating pressure, the pressure sensors may monitor the pressure and ensure that the pressure can be maintained (e.g., without leaking) for a predetermined period of time. Once the system determines that it is capable of maintaining the operating pressure, the system may provide an output to the user indicating that the system has passed the pressure integrity test. The system may then be pressurized.

[0192] In some embodiments, the predetermined amount of time may be about 10 seconds, 20 seconds, 30 seconds, 60 seconds, or 90 seconds. In some embodiments, an initial pressurization cycle of about 20 seconds may be followed by a pressure maintenance cycle of about 1 minute. In some embodiments, the system may require that the pressure be normally maintained within a range of about + / -1 psi, + / -5 psi, or + / -10 psi. In some embodiments, the range may be greater than + / -0.5 psi, + / -1 psi, + / -5 psi, or + / -10 psi. In some embodiments, the range may be less than + / -1 psi, + / -5 psi, + / -10 psi, or + / -15 psi.

[0193] In some embodiments, if the system indicates a failure of one or more components of the integrity test, the system may display an indication to the user regarding the connection or component that must be corrected or replaced. In some embodiments, the user may remove the defective disposable assembly and insert a replacement disposable assembly.

[0194] At block 1314, in some embodiments, a primer solution may be passed through a disposable assembly. In some embodiments, passing a primer solution (e.g., a buffer fluid) through the assembly before passing the cell suspension through the system may help prevent the cells of the cell suspension from sticking to and / or being damaged by the internal surfaces of the system's flow paths. In some embodiments, the primer solution passed through the system may originate from a buffer input bag, such as buffer input bag 108 in system 100.

[0195] In some embodiments, the user may provide the buffer input bag to the system in the same or similar manner that the user may provide the cell suspension input bag to the system before performing the integrity check, such as by suspending the bag from a hook and / or by fluidly connecting the bag to an inlet of the system (e.g., an inlet of a preparation vessel of the system).

[0196] In some embodiments, the system may be configured to partially or fully automatically cause the primer solution to flow through the disposable assembly, including by providing and monitoring gas pressure and forcing fluid along a flow path and / or by opening and closing valves, clamps, and / or ports as appropriate. In some embodiments, the system may be configured to receive input from a user (e.g., input implemented at a user interface device of the system, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals to components of the system to cause the flow of primer solution through the disposable assembly.

[0197] For example, the system may pressurize gas in the system's preparation vessel by generating and transmitting a signal to the system's pressure control module, activating one or more gas pumps, opening one or more valves, and / or operating one or more regulators, etc., to cause pressurized gas to flow into the vessel and force the primer solution to flow through the disposable assembly. Additionally, the system may generate and transmit a signal to one or more valves (e.g., solenoid valves), clamps, or other flow control mechanisms disposed along the system's flow path to open a flow path and allow the primer solution to flow through the disposable assembly. For example, any of the valves, clamps, or fittings shown in system 1000 in FIG. 10 may be configured to be electronically controlled so as to be automatically actuated by the system. Furthermore, the system may monitor data generated by one or more sensors (e.g., flow sensor 912) and automatically determine when the primer solution has completed the step of flowing through the disposable assembly, thereby performing additional actions and / or notifying the user that the priming process is complete.

[0198] In some embodiments, the primer solution may flow into an output bag, such as buffer output bag 114, and may then be removed and / or disposed of by the user.

[0199] At block 1316, in some embodiments, the cell suspension may be passed into the preparation vessel. In some embodiments, the cell suspension may be passed into the preparation vessel after the primer solution has been passed out of the preparation vessel, through the remainder of the disposable assembly, and into an output bag. In some embodiments, the system may be configured to partially or fully automatically flow the primer solution out of the input bag and into the preparation vessel, including by controlling any one or more valves, clamps, pumps, regulators, or other pressure and / or flow control mechanisms in the same or similar manner as described with reference to step 1314 above.

[0200] In some embodiments, the system may be configured to receive input from a user (e.g., input executed at a user interface device of the system, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals to components of the system to cause the flow of primer solution through the disposable assembly. In some embodiments, the system may be configured to automatically cause the cell suspension to enter the preparation vessel in response to detecting completion of the primer process.

[0201] Additionally, the system may monitor data generated by one or more sensors (e.g., flow sensors) and automatically determine when the cell suspension has completed the step of flowing into the preparation vessel, and therefore perform additional actions and / or notify the user that the process is complete.

[0202] At block 1318, in some embodiments, the cell suspension may be prepared for passage through a constriction cartridge while the cell suspension is in the preparation vessel. In some embodiments, preparing the cell suspension for passage through the constriction cartridge may include performing one or more manual or automated actions to alter (or maintain) one or more properties of the cell suspension. For example, the cell suspension may be prepared for passage through the constriction cartridge in some embodiments by cooling the cell suspension, heating the cell suspension, agitating the cell suspension, and / or applying pressure to the cell suspension. In some embodiments, the preparation process may involve performing one of temperature control, pressure control, and / or agitation simultaneously, while in some embodiments, it may involve performing one or more of them sequentially. For example, the temperature control process may take significantly longer than the pressurization process, and therefore, the temperature control process may in some embodiments be performed before the pressurization process (e.g., the system may begin pressurizing the vessel only once the system determines that the cell suspension has reached a target temperature range). Although the discussion herein may consider steps of the preparation process to be performed simultaneously, they may, in some embodiments, be performed simultaneously and / or in any sequential order.

[0203] In block 1320, in some embodiments, preparing the cell suspension for passage through the constriction cartridge may include applying pressure to the cell suspension. As discussed above, pressurized gas may be used to apply pressure to a liquid in a system's flow path, urging the liquid to flow along the flow path. In some embodiments, when the cell suspension is stored inside a preparation vessel, the space above the cell suspension in the preparation vessel may be filled with pressurized sterilizing gas to apply a force to the liquid that can be used to urge it toward an outlet at the bottom of the preparation vessel. In an example of system 100, preparation vessel 600 may be partially filled with the cell suspension, and pressurized sterilizing gas, such as nitrogen, may then be flowed into the vessel through one of vessel inlets 604. In an example of system 1000, pressurized gas may flow into vessel 1016 through connector 1062 to apply pressure to the cell suspension in the vessel.

[0204] In some embodiments, the system may be configured to receive input from a user (e.g., input implemented on a user interface device of the system, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals to components of the system to cause gas pressurization inside the preparation vessel. In some embodiments, the system may be configured to automatically cause gas pressurization inside the preparation vessel in response to detecting that the cell suspension has fully flowed into the vessel.

[0205] In some embodiments, the system may be configured to cause pressurization of gas inside the preparation vessel partially or fully automatically, including by controlling any one or more valves, pumps, regulators, or other pressure control mechanisms in the same or similar manner as described above with reference to step 1314. In some embodiments, the system may monitor the pressure inside the preparation vessel or may monitor the gas pressure elsewhere in the pressure control assembly to determine whether the pressure inside the preparation vessel needs to be increased, decreased, or maintained to achieve the desired operating pressure. In some embodiments, the system may use a pump to increase the pressure from a low-pressure source to a desired pressure (e.g., operating pressure). In some embodiments, the operating pressure may be about 20 psi, about 30 psi, about 50 psi, about 70 psi, about 90 psi, about 110 psi, or about 130 psi. In some embodiments, the operating pressure may be greater than 10 psi, 20 psi, 50 psi, 70 psi, 90 psi, 110 psi, or 130 psi. In some embodiments, the operating pressure may be less than 20 psi, 50 psi, 70 psi, 90 psi, 110 psi, 130 psi, or 150 psi.

[0206] While block 1320 above discusses applying pressure to the cell suspension, alternative or additional techniques may be used in some embodiments to cause the flow of the cell suspension out of the preparation vessel and / or into and through the constriction cartridge. For example, in some embodiments, as an alternative to or in addition to pressurizing a gas within the preparation vessel, the cell suspension may be caused to flow out of the preparation vessel and / or into and through the constriction cartridge by gravity, by vacuum force, by centrifugation, and / or by force applied by a pump.

[0207] At block 1322, in some embodiments, preparing the cell suspension for passage through the constriction cartridge may include agitating the cell suspension. In some embodiments, agitating the cell suspension may ensure uniform distribution of cells throughout the suspension and prevent cells from being non-uniformly dispersed in the cell suspension before the suspension is passed through the constriction cartridge. In some embodiments, agitation may be achieved by shaking or vibrating the preparation vessel. In an example of system 100, preparation vessel 600 may be agitated due to shaking of shaker plate 304, as shown in FIG. 3 . In some embodiments, a user may be able to set the agitation rate, intensity, and / or duration. In some embodiments, the system may automatically determine and / or set the agitation rate, intensity, and / or duration. In some embodiments, a user may be able to select whether agitation is used as part of the cell suspension preparation process.

[0208] In some embodiments, the system may be configured to receive input from a user (e.g., input implemented on a user interface device of the system, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals that cause components of the system to agitate the cell suspension inside the preparation vessel. In some embodiments, the system may be configured to automatically cause agitation of the cell suspension inside the preparation vessel in response to detecting that the cell suspension has completely flowed into the vessel.

[0209] In some embodiments, the system may be configured to partially or fully automatically cause agitation of the cell suspension inside the preparation vessel, including by controlling any one or more shaker plates, vibration devices, kneading devices, sonic agitation devices, peristaltic pump devices, gas / diaphragm devices, or other mechanisms configured to cause shaking / vibration of the preparation vessel and / or agitation / circulation of the cell suspension inside the vessel. In some embodiments, methods other than agitation, such as a compartmentalized intake (e.g., small aliquots of input may be added over time to keep cells in suspension), may also be used to prevent cells from falling out of suspension, resuspend cells, and / or homogenize the cell suspension. In an example of system 100, the system may be configured to automatically electronically control motor 302 and / or belt drive 306 to control the agitation of shaker plate 304.

[0210] In block 1324, in some embodiments, preparing the cell suspension for passage through the constriction cartridge may include controlling the temperature of the cell suspension. In some embodiments, controlling the temperature of the cell suspension may include cooling the cell suspension, heating the cell suspension, or ensuring that the temperature of the cell suspension remains unchanged. In some embodiments, the payload delivery process may be most effective at a specific predetermined temperature or temperature range, and the system may therefore be configured to be able to heat or cool the cell suspension to that predetermined temperature or temperature range before flowing the suspension through the constriction cartridge. In some embodiments, the system may be configured to heat and / or cool the cell suspension to have a temperature greater than about 0, 1, 2, 3, 4, 5, 10, 20, 30, 35, 37, or 40 degrees Celsius. In some embodiments, the system may be configured to heat and / or cool the cell suspension to have a temperature less than about 1, 2, 3, 4, 5, 10, 20, 30, 35, 37, 40, or 45 degrees Celsius. In some embodiments, the system may be configured to bring the cell suspension from room or storage temperature to the target temperature range within 30 minutes or less, 60 minutes or less, 90 minutes or less, or 120 minutes or less. In some embodiments, the system may be configured to operate at room temperature. In some embodiments, the system may be configured to operate at about 22-24 degrees Celsius, about 21-25 degrees Celsius, about 20-26 degrees Celsius, or about 18-28 degrees Celsius.

[0211] In some embodiments, the system may be configured to receive input from a user (e.g., input implemented on a user interface device of the system, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals that cause components of the system to adjust the temperature of the cell suspension inside the preparation vessel. In some embodiments, the system may be configured to automatically adjust the temperature of the cell suspension inside the preparation vessel in response to detecting that the cell suspension has fully flowed into the vessel.

[0212] In some embodiments, the system may be configured to regulate the temperature of the cell suspension inside the preparation vessel partially or fully automatically, including by controlling any one or more suitable heating or cooling devices, optionally in conjunction with any suitable temperature sensors as discussed elsewhere herein. In some embodiments, the temperature control device may comprise one or more forced air heaters, one or more forced air coolers, one or more thermoelectric cooling devices (e.g., Peltier coolers), one or more resistive heating devices, or the like. In examples of system 100, the temperature control device may be part of temperature control module 118. In some embodiments, the temperature control device of the system may be in physical contact with a preparation vessel housing, such as preparation vessel housing 110, or the temperature control device may be otherwise configured to heat and / or cool the preparation vessel housing. By heating and / or cooling the preparation vessel housing while the preparation vessel is contained within the housing, heat may be transferred to or from the preparation vessel and, in turn, to or from the cell suspension inside the preparation vessel, thereby achieving heating and / or cooling of the cell suspension.

[0213] In some embodiments, to achieve a predetermined target temperature, the system may be configured to continuously monitor the temperature of the cell suspension while the temperature control process is ongoing. For example, system 100 may be configured to continuously monitor the temperature associated with the cell suspension via temperature sensor 914, which may be attached to the exterior surface of preparation vessel 600. Temperature sensor 914 may transmit data to temperature control module 118 and / or one or more other processors of system 100, and based on the received data, system 100 may determine whether the target temperature has been achieved, whether the temperature control process needs to continue, and whether a heating or cooling device being used in the temperature control process needs to be adjusted (e.g., to a higher or lower temperature).

[0214] In some embodiments, one or more temperature sensors of the system may be configured to read both a PID (proportional-integral-derivative) temperature and an NTC (negative temperature coefficient) temperature and calculate an effective temperature based on both of those readings. The system may determine whether the calculated effective temperature is within a target temperature range. If the effective temperature is not within the target temperature range, the system may adjust one or more heating or cooling elements and / or wait a predetermined amount of time before calculating a new effective temperature based on the new PID and NTC temperature readings and checking whether the new effective temperature is within the target range. If the effective temperature is determined to be within the target range, the system may indicate to the user that the temperature control process is complete and / or terminate the temperature adjustment process. In some embodiments, if a predetermined amount of time has elapsed and the effective temperature still has not reached the target temperature range, the system may return a timeout error and indicate to the user that the temperature control process has failed.

[0215] In some embodiments, rather than heating or cooling the cell suspension as part of the preparation process inside the preparation vessel, the cell suspension may instead be heated or cooled before entering the preparation vessel. For example, the cell suspension may, in some embodiments, be heated or cooled while inside an input bag or another portion of the flow path upstream of the preparation vessel. Alternatively, or in addition, the cell suspension may, in some embodiments, be heated or cooled to a target temperature range as part of a batch temperature control process in which a large volume of cell suspension suitable for use in a multiple payload delivery process is heated or cooled at once. By heating or cooling the cell suspension in a large batch volume, the overall throughput time for a multiple payload delivery process may be improved because time may be saved by not having to individually (and sequentially) heat or cool each small batch of cell suspension. In some embodiments, heating or cooling may be performed both before the cell suspension arrives in the preparation vessel and during the time the cell suspension is in the preparation vessel. For example, coarse adjustments to the suspension temperature may be made before the preparation vessel, while fine adjustments to the temperature may be made while the suspension is in the preparation vessel.

[0216] In step 1326, in some embodiments, a payload may be contacted with the cell suspension while it is in the preparation vessel. In some embodiments, the payload may include one or more peptides, nucleic acids, proteins, carbohydrates, lipids, small molecules, complexes, and / or nanomaterials, which may be included as part of the suspension. In some embodiments, this may occur before, during, or after any one or more of the cell suspension preparation steps discussed with respect to steps 1318-1324 above. In either case, note that in some embodiments, the payload may be contacted with the cell suspension before the cell suspension is passed through the constriction cartridge. On the other hand, in some embodiments, such as those discussed below with reference to step 1330, the payload may be contacted with the cell suspension only after the cell suspension has passed through the constriction cartridge. In embodiments in which the payload is mixed with the cell suspension before passage through the constriction cartridge, the payload may be passed through the constriction cartridge along with the cell suspension.

[0217] While in the preparation vessel, the payload is contacted with the cell suspension. In some embodiments, the payload may be mixed as part of the cell suspension in an input bag, or the payload may be inserted separately into the preparation vessel through an inlet to the preparation vessel. In some embodiments, the payload may be provided in a dedicated input bag and may flow into the preparation vessel and mix with the cell suspension. In any of these embodiments, the flow of the payload through the system may be electronically controlled by the system (e.g., automatically, by opening a valve, applying pressure, etc.) in any of the same ways as discussed above with respect to controlling the flow of the cell suspension itself.

[0218] In step 1328, in some embodiments, the cell suspension may be passed through a constriction cartridge. As discussed elsewhere herein, the cell suspension may be forced under pressure to flow through a constriction cartridge, which may cause the cell suspension to flow through one or more cell-narrowing microfluidic channels and / or one or more cell-narrowing filters. As cells of the cell suspension are forced through the constrictions of the cell-narrowing microfluidic channels and / or cell-narrowing filters, the cell membranes may be perturbed, and the perturbation may facilitate entry of the payload into the cells.

[0219] In some embodiments, the system may be configured to partially or fully automatically flow the cell suspension out of the preparation vessel and into and through the constriction cartridge, including by controlling any one or more valves, clamps, pumps, regulators, or other pressure and / or flow control mechanisms in the same or similar manner as described above with reference to step 1314. In some embodiments, the gas inside the preparation vessel is pressurized, and the system may electronically open an outlet of the preparation vessel to allow gas pressure to urge the cell suspension to flow out of the preparation vessel. In an example of system 100, a valve associated with outlet 606 may be opened. In an example of system 1000, a valve associated with connector 1018 may be opened.

[0220] In some embodiments, the system may be configured to receive input from a user (e.g., input implemented on a system user interface device, such as touchscreen 120 of system 100) and, in response, generate and transmit one or more electronic signals to components of the system to cause the flow of the cell suspension out of the cell preparation container and into and through the constriction cartridge. In some embodiments, the system may be configured to automatically cause the flow of the cell suspension out of the cell preparation container and into and through the constriction cartridge in response to detecting completion of a primer cell suspension preparation process. In some embodiments, the system may be configured to cause the cell suspension to flow into and through the constriction cartridge over a period of about 30 seconds, 1 minute, 2 minutes, or 3 minutes. In some embodiments, the period may be greater than 10 seconds, 30 seconds, 1 minute, 2 minutes, or 3 minutes. In some embodiments, the period may be less than about 30 seconds, 1 minute, 2 minutes, 3 minutes, or 5 minutes. In some embodiments, the systems and techniques disclosed herein may allow passage of up to about 1 billion cells / minute, 1.5 billion cells / minute, or 2 billion cells / minute, or more, through a constriction cartridge.

[0221] Additionally, the system may monitor data generated by one or more sensors (e.g., flow sensors) and automatically determine when the cell suspension has completed the step of flowing into and through the constriction cartridge, and thus perform additional actions and / or notify the user that the process is complete. In an embodiment of system 100, flow sensor 912 may be configured to optically monitor the tubing extending from preparation vessel 600 to the constriction cartridge and determine when the flow of the cell suspension through the tubing has stopped.

[0222] In step 1330, in some embodiments, the payload may be contacted with the cell suspension following passage of the cell suspension through the constriction cartridge. As discussed above with respect to step 1326, in some embodiments, the payload may be contacted with the cell suspension before the cell suspension is passed through the constriction cartridge. On the other hand, in some other embodiments, the payload may be contacted with the cell suspension only after the cell suspension has passed through the constriction cartridge. In some embodiments, the payload may actually enter the cells of the cell suspension even when they are contacted only after the cell suspension has passed through the constriction cartridge. That is, the perturbation induced in the cell membrane may allow effective payload entry for a period of time after the perturbation is induced by the constriction.

[0223] In some embodiments, where the payload is contacted with the cell suspension only after the cell suspension has passed through the constriction cartridge, an independent source of payload suspension may be flowed into the same channel, reservoir, or container as the cell suspension. For example, in some embodiments, the payload may be mixed with the cell suspension while or after the cell suspension flows into an output bag, such as output bag 112 of system 100. Thus, in some embodiments, a portion of the cell suspension may contact the payload (e.g., in the output bag) after the suspension has flowed through the constriction cartridge, but this may occur while the portion of the cell suspension is still in the preparation container and / or has not yet flowed through the constriction cartridge. In any of these embodiments, the flow of the payload through the system to contact the cell suspension may be electronically controlled by the system (e.g., automatically, by opening a valve, applying pressure, etc.) in any of the same ways as discussed above with respect to controlling the flow of the cell suspension itself.

[0224] In some embodiments, in step 1332, the cell suspension may be passed into an output container. In an example of system 100, the output container may be output bag 112, which, after being filled, may be disconnected and removed from system 100 for further processing by a user.

[0225] In some embodiments, one or more computing devices of the intracellular payload delivery system may monitor one or more characteristics or properties of the cell suspension during all or a portion of method 1300. In some embodiments, the system may monitor elapsed times, such as the elapsed time for a primer process, the elapsed time for an integrity check, the elapsed time for a cell suspension preparation process (e.g., cooling, air pressurization, etc.), the elapsed time for the cell suspension to flow through the constriction cartridge, the total elapsed time for the entire process, and / or the elapsed time for a combination of any two or more of any of the above, during all or a portion of method 1300. In some embodiments, the system may monitor the pressure of the system over time during any one or more portions of the overall process. In some embodiments, the system may monitor the temperature of the cell suspension over time during any one or more portions of the overall process.

[0226] In some embodiments, any of the monitored characteristics or properties may be stored locally to the system as part of one or more log files or databases, may be transmitted to a remote electronic device for storage, processing, or display, and / or may be displayed on a display at the local or remote electronic system (e.g., display 120 of system 100). User Interface

[0227] 14A-14V illustrate a user interface 1402 for controlling a tabletop system for delivering a payload to a cell, according to some embodiments. In some embodiments, the user interface 1400 may be displayed by any suitable display device, such as the display 120 of the system 100, which may be located locally to the intracellular payload delivery system (e.g., integrated into the body of the device or attached to the device via wired electronic communication) or remotely from the intracellular payload delivery system (e.g., configured to communicate with the system via wireless electronic communication). In some embodiments, the user interface 1400 may be configured for use with a touchscreen display, such that a user may touch or tap with a finger or stylus on displayed buttons or icons of the interface. In some embodiments, the user interface 1400 may be configured for use with a non-touchscreen display, such that a user may navigate and interact with the interface using a mouse, keyboard, keypad, buttons, pressure-sensitive devices, knobs, joystick, motion sensing, voice control, and / or other input devices.

[0228] 14A-14V, screens 1402A-1402V are displayed on display 1400, which may be any local or remote touchscreen display and may share some or all characteristics in common with display 120 of system 100. As shown and described below, screens 1402A-1402V may be displayed during various portions of an intracellular payload delivery process, such as method 1300 described above. As the intracellular payload delivery process progresses, the intracellular payload delivery system may receive various inputs from a user and display various instructions, alerts, and measurements to the user via user interface 1400.

[0229] 14A illustrates a screen 1402A that is displayed on a display 1400. The screen 1402A includes a boot-up message 1404, which may be any graphical and / or textual alert that may be displayed to the user to indicate that the intracellular payload delivery system is booting up.

[0230] 14B illustrates screen 1402B, which is displayed on display 1400. Screen 1402B may be displayed when the intracellular payload delivery system is idle, such as after boot-up or when the system is not currently performing any intracellular payload delivery processes. Screen 1402B may include an intracellular payload delivery process start icon 1406, which may be tapped or clicked by a user to generate an input to instruct the system to begin the intracellular payload delivery process.

[0231] 14C illustrates a screen 1402C displayed on the display 1400. The screen 1402C may include various options for configuring or preparing the intracellular payload delivery process, including setting various settings to be used during the process. For example, the screen 1402C may allow the user to instruct the device whether to use a cooling process, whether to use an agitation process, and the gas pressure to be used for the process.

[0232] Screen 1402C may include a cooling process selection icon 1406 that may allow the user to tap or click the appropriate icon and generate input to instruct the system to use a cooling (or heating) process, or alternatively, not use a cooling (or heating) process. In some embodiments, one or more additional icons or user interface elements may be displayed to allow the user to perform input to generate instructions for setting a target temperature or target temperature range. Once the user makes a selection using icon 1406, the setting may be saved and applied to one or more future payload delivery processes performed by the system.

[0233] Screen 1402C may include an agitation process selection icon 1408 that may allow the user to tap or click the appropriate icon and generate input to instruct the system to use the agitation process, or alternatively, not use the agitation process. In some embodiments, one or more additional icons or user interface elements may be displayed to allow the user to perform input to generate instructions for setting the agitation rate, intensity, and / or duration. Once the user makes a selection using icon 1408, the setting may be saved and applied to one or more future payload delivery processes performed by the system.

[0234] Screen 1402C may include pressure selection icons 1410, which may allow the user to tap or click on the appropriate icon and generate input to instruct the system regarding the gas pressure to be used for the payload delivery process. Screen 1402C may also include a pressure setting display 1412, which may display the pressure currently selected by the user, such as by displaying the pressure in pounds per square inch. Once the user makes a selection using icon 1410, the setting may be saved and applied to one or more future payload delivery processes performed by the system.

[0235] Screen 1402C may include a stop icon 1416 that may abort the payload delivery process or act as a back button and cause the system to display the previously displayed screen. Screen 1402C may also include a save / continue icon 1414 that may be tapped or clicked by the user to save any settings or inputs made on the current screen and proceed to the next screen and / or next phase in the payload delivery process.

[0236] FIG. 14D illustrates screen 1402D, displayed on display 1400. Screen 1402D may be a screen displayed to instruct a user to deploy and / or attach all or a portion of a disposable assembly, such as a sensor assembly. In some embodiments, the disposable assembly may be associated with a code, identification number, barcode, QR code, or the like, which may be used to ensure single-use and / or regulatory compliance. In some embodiments, the system may prompt the user to enter or otherwise present the code or identification information of the disposable assembly to the system (e.g., via display 1400) so that the system may verify the disposable assembly. In some embodiments, the system may display instructions for disposable assembly deployment each time a payload delivery process is performed, such as when the disposable assembly is configured for one-time use only. In some embodiments, the system may read data from one or more sensors to determine whether the disposable assembly is already attached and may display attachment / deployment instructions only if the assembly is not already attached.

[0237] As shown, screen 1402D may include sensor assembly instructions 1418, which may be any graphical and / or textual instructions regarding how to position and install the sensor assembly. Similarly, screen 1402D may include sensor assembly positioning image 1420, which may be an image or video illustrating one or more portions of the instructed positioning / installation process. In the example of FIG. 14D, the sensor assembly instructions instruct the user to extend tubing from the preparation vessel through the foam sensor, clip the constriction cartridge into the seat, and plug the sensor assembly's electronic connector into the system's electronic interface. In some embodiments, the instructions displayed may depend on settings selected by the user in one or more previous screens.

[0238] Screen 1402D may include a next icon 1422 and a previous icon 1424 that may be tapped or clicked by the user to move to the next or previous screen, respectively. In some embodiments, the system may automatically display the next screen upon detecting that placement of the sensor assembly is complete.

[0239] 14E illustrates screen 1402E displayed on display 1400. Screen 1402E may be a screen displayed to instruct a user to deploy and / or install all or a portion of a disposable assembly, such as a preparation vessel assembly, a filter assembly, and / or a gas assembly. In some embodiments, the system may display instructions for disposable assembly deployment each time a payload delivery process is performed, such as when the disposable assembly is configured for one-time use only. In some embodiments, the system may read data from one or more sensors to determine whether the disposable assembly is already installed and may display installation / installation instructions only if the assembly is not already installed.

[0240] As shown, screen 1402E may include disposable assembly instructions 1426, which may be any graphical and / or textual instructions regarding how to dispose and attach the sensor assembly. Similarly, screen 1402E may include disposable assembly disposition images 1428, which may be images or videos illustrating one or more portions of the instructed disposition / attachment process. In the example of FIG. 14E, the disposable assembly instructions instruct the user to insert the preparation vessel into its housing, connect the filter to the system, and connect a pressurized sterile gas source and vent line to the filter assembly. In some embodiments, the instructions displayed may depend on settings selected by the user in one or more previous screens.

[0241] Like screen 1402D, screen 1402E may include a next icon 1422 and a previous icon 1424 that may be tapped or clicked by the user to move to the next or previous screen, respectively. In some embodiments, the system may automatically display the next screen upon detecting that disposable assembly placement is complete.

[0242] 14F illustrates screen 1402F displayed on display 1400. Screen 1402F may include integrity test instructions 1430, which may include any graphical and / or written instructions for a user to prepare the system for an integrity test, such as a cartridge integrity test. In the example of FIG. 14F, the integrity test instructions instruct the user to ensure that certain input and output valves of the system are closed so that the system can be pressurized without gas leaking into other system components.

[0243] Like screen 1402D, screen 1402F may include a next icon 1422 and a previous icon 1424 that may be tapped or clicked by the user to move to the next or previous screen, respectively. In some embodiments of screen 1402F, selecting next icon 1422 may cause the system to initiate an integrity test by flowing pressurized gas into the system's flowpath.

[0244] FIG. 14G illustrates screen 1402G displayed on display 1400. Screen 1402G may be a screen displayed by the system while one or more integrity tests are being performed. During an integrity test of one or more system components, pressurized gas may be flowed into one or more system components, and the pressure inside the one or more system components may be monitored. As discussed above, if the target pressure can be achieved and maintained, the system may determine that the integrity test has passed. If the target pressure cannot be achieved or maintained for at least a predetermined period of time, the system may determine that the integrity test has failed, and one or more settings may be required to be changed and / or one or more system components may be required to be adjusted or replaced.

[0245] Screen 1402G may include a dynamic pressure indicator 1432, which may display a dynamic indication of the current internal pressure of the system component or components undergoing an integrity test (e.g., preparation vessel, constriction cartridge, etc.). Screen 1402G may include a dynamic elapsed time indicator 1434, which may display a dynamic indication of the current elapsed time for the integrity test being performed.

[0246] In some embodiments, the system may automatically display the next screen once it detects that the integrity test is complete.

[0247] 14H illustrates screen 1402H, which is displayed on display 1400. Screen 1402H may be displayed if the system determines that the integrity test was successful in that all components passed the integrity test. Screen 1402H may include an integrity test success message 1436, which may be any graphical and / or textual indication to alert the user that the integrity test passed.

[0248] Screen 1402H may include a next icon 1422 that may be tapped or clicked by the user to move to the next screen. In some embodiments of screen 1402H, the system may automatically display the next screen a predetermined period of time after beginning to display screen 1402H.

[0249] 14I illustrates screen 1402I, which is displayed on display 1400. Screen 1402I may be displayed if the system determines that the integrity test was unsuccessful in that one or more components did not pass the integrity test. Screen 1402H may include an integrity test failed message 1438, which may be any graphical and / or textual indication to alert the user that the integrity test did not pass.

[0250] Screen 1402I may further include component replacement instructions 1440, which may be any graphical and / or textual instructions to indicate to the user that one or more system components need to be adjusted or replaced and / or one or more system settings need to be changed before proceeding (such as by retrying the integrity test with a new disposable assembly). In the example of FIG. 14I, component replacement instructions 1440 instruct the user to remove a portion of the disposable assembly and obtain a new one.

[0251] Screen 1402I may include a next icon 1422 that may be tapped or clicked by the user to move to the next screen. In some embodiments of screen 1402I, the system may display instructions for disposing a new component that the user has been instructed to remove after the user taps or clicks next icon 1422. In some embodiments, the system may automatically display new instructions for component disposition in response to the system detecting that a failed component has been removed or disconnected from the system.

[0252] 14J illustrates screen 1402J, which is displayed on display 1400. Screen 1402J may be displayed following a successful integrity test. For example, screen 1402J may be displayed following screen 1402H. In some embodiments, screen 1402J may provide instructions for a user to arrange / attach input sources for fluidic materials, such as buffers / primers, cell suspensions, and / or payload materials, and flow them through the system during the payload delivery process. For example, a user may be instructed to suspend an input bag from a hook on the system and attach a tubing connector to the inlet of a preparation vessel.

[0253] As shown, screen 1402J may include fluid input source assembly instructions 1442, which may be any graphical and / or textual instructions regarding how to arrange and attach one or more fluid input sources. Similarly, screen 1402J may include fluid input source arrangement images 1444, which may be images or videos illustrating one or more portions of the instructed arrangement / attachment process. In the example of FIG. 14J, the fluid input source arrangement instructions instruct the user to hang the source and primer input bag from hooks and attach tubing to the inlet of the preparation vessel. In some embodiments, the instructions displayed may depend on settings selected by the user in one or more previous screens.

[0254] Screen 1402J may include a next icon 1422 that may be tapped or clicked by the user to move to the next screen. In some embodiments, the system may automatically display the next screen upon detecting that placement of the fluid input source is complete.

[0255] FIG. 14K illustrates screen 1402K, displayed on display 1400. Screen 1402K may be an interface that allows a user to select whether the system should use a priming process as part of the payload delivery process. Screen 1402K may include a priming process selection icon 1446 that may allow a user to generate an input to instruct the system regarding whether to use a priming process by tapping or clicking the appropriate icon and passing a primer solution through the system's flow paths before passing a cell suspension through the system's flow paths. In some embodiments, screen 1402K may further include one or more icons or other user interface objects to allow a user to enter parameters or settings for the priming process, such as the source of the primer solution or the pressure or temperature to be used for the priming process. Once the user makes a selection using icon 1446, the settings may be saved and applied to one or more future payload delivery processes performed by the system.

[0256] FIG. 14L illustrates screen 1402L, which is displayed on display 1400. Screen 1402L, in some embodiments, may be displayed in response to a user indicating that a priming process should be performed using icon 1446. Screen 1402L may include prime preparation instructions 1448, which may include any graphical and / or written instructions for a user to prepare the system for the priming process. In the example of FIG. 14L, prime preparation instructions 1448 instruct the user to fill the preparation vessel with primer / buffer solution, ensure that the valves are in the proper orientation, and ensure that the valve to the cell suspension source bag is closed. Note that in some embodiments, these or other steps that the system instructs the user to perform may instead be performed automatically by the system, such as by electronically actuating valves.

[0257] Like screen 1402D, screen 1402L may include a next icon 1422 and a previous icon 1424 that may be tapped or clicked by the user to move to the next or previous screen, respectively. In some embodiments of screen 1402L, selecting next icon 1422 may cause the system to initiate a priming process by actuating one or more valves or other system components and causing primer / buffer solution to flow through the system's flow paths.

[0258] 14M illustrates screen 1402M, which is displayed on display 1400. Screen 1402M may be displayed once the system has completed the priming process. Screen 1402M may include a dynamic pressure indicator 1450, which may display a dynamic indication of the current internal pressure of one or more system components or contents (e.g., preparation vessel, constriction cartridge, etc.) during the priming process. In some embodiments, screen 1402M may include a dynamic temperature indicator 1452, which may display a dynamic indication of the current temperature of one or more system components or contents (e.g., the calculated effective temperature of the primer solution) during the priming process. In some embodiments, screen 1402M may include a dynamic elapsed time indicator (not shown), such as dynamic elapsed time indicator 1434, which may display a dynamic indication of the current elapsed time during the priming process.

[0259] In some embodiments, the system may automatically display the next screen once it detects that the priming process is complete.

[0260] 14N illustrates screen 1402N, which is displayed on display 1400. Screen 1402N may be displayed when the system detects that the priming process is complete. For example, when a flow sensor (e.g., flow sensor 912 of system 100) detects that primer / buffer solution is no longer flowing through one or more tubes of the system, the system may determine that the priming process is complete and may display screen 1402N. Screen 1402N, in some embodiments, may prompt the user to confirm that the priming process is complete.

[0261] Screen 1402N may include a priming completion confirmation icon 1454 that may be tapped or clicked by the user to indicate whether the priming process is complete. In some embodiments, indicating that the priming process is complete (e.g., by tapping the "Yes" icon) may cause the system to proceed to the next step and display the next screen. In some embodiments, indicating that the priming process is not complete may cause the system to continue the priming process for a predetermined period of time until the user indicates otherwise or until the system again detects that the priming process is complete. In some embodiments, confirming that the priming process is complete may cause the system to actuate one or more valves or other system components to close primer / buffer solution flow paths and / or open flow paths for the cell suspension and may cause the system to proceed to the next screen.

[0262] FIG. 14O illustrates screen 1402O, displayed on display 1400. Screen 1402O may include cell suspension flow process setup instructions 1456, which may include any graphical and / or written instructions for a user to prepare the system for a cell suspension flow process in which the cell suspension is flowed through a preparation vessel, a constriction cartridge, and / or other system components. In the example of FIG. 14O, cell suspension flow process setup instructions 1456 instruct the user to fill a preparation vessel with the cell suspension and press a button to begin the cell suspension flow process. Note that in some embodiments, these or other steps that the system instructs the user to perform may instead be performed automatically by the system, such as by electronically actuating a valve.

[0263] Screen 1402O may include a start cell suspension flow process icon 1458 that may be tapped or clicked by a user to signal an instruction to the system to begin the cell suspension flow process by actuating one or more valves or other system components, flowing the cell suspension into and through the preparation vessel, preparing it while in the preparation vessel, flowing it into and through the constriction cartridge, flowing it into an output bag, etc. In some embodiments, selecting icon 1458 to start the cell suspension flow process may also cause the system to begin monitoring one or more characteristics of the system and / or cell suspension, such as elapsed time, pressure, temperature, and / or agitation state, and may cause the system to advance to the next screen.

[0264] FIG. 14P illustrates screen 1402P, displayed on display 1400. Screen 1402P may include an agitation process selection icon 1460, which may allow a user to tap or click an appropriate icon and generate input to instruct the system to use the agitation process, or alternatively, not use the agitation process. In some embodiments, one or more additional icons or user interface elements may be displayed, allowing a user to execute input to generate instructions for setting the agitation rate, frequency, intensity, amplitude, and / or duration. Once a user makes a selection using icon 1409, the settings may be saved and applied to one or more future payload delivery processes performed by the system. In some embodiments, agitation selection icon 1460 may differ from agitation selection icon 1409 on screen 1402C in that icon 1409 may be used to set general system settings that apply by default to all payload delivery processes, while icon 1460 may be used to set specific settings that apply only to the current payload delivery process.

[0265] In some embodiments, selecting any of the agitation selection icons 1460 may cause the system to display the next screen, as shown, and proceed with the cell suspension flow process with or without agitation.

[0266] 14Q illustrates screen 1402Q, which is displayed on display 1400. Screen 1402Q may, in some embodiments, be displayed while the cell suspension is being prepared inside the preparation vessel, such as by being cooled, stirred, and / or subjected to increased gas pressure.

[0267] Screen 1402Q may include a dynamic pressure indicator 1462, which may display a dynamic indication of the current internal pressure of a system component or components (e.g., preparation vessel, constriction cartridge, etc.) during the cell suspension preparation process. Screen 1402Q may include a dynamic temperature indicator 1464, which may display a dynamic indication of the current temperature of one or more system components or contents (e.g., the calculated effective temperature of the cell suspension) during the cell suspension preparation process. Screen 1402Q may include a dynamic process timer indicator 1466, which may display a dynamic indication of the elapsed time for the current payload delivery process, cell suspension flow process, and / or cell suspension preparation process.

[0268] In some embodiments, the system may automatically advance to the next screen after a predetermined period of time once the system detects that the preparation process is complete (such as by detecting that the system's dynamic pressure has reached and / or maintained a target pressure and / or that the system's dynamic temperature target pressure has reached and / or maintained).

[0269] FIG. 14R illustrates screen 1402R, which is displayed on display 1400. Screen 1402R, in some embodiments, may be displayed after the cell suspension preparation process is completed. Screen 1402R may also include post-preparation instructions 1468, which may include any graphical and / or written instructions for a user to prepare the system for the remainder of the cell suspension flow process following preparation of the cell suspension in the preparation vessel, such as by adjusting valves or other system components prior to the cell suspension being forced to flow under pressure through the constriction cartridge. In the example of FIG. 14R, post-preparation instructions 1468 instruct the user to ensure that valves are in the proper orientation and that the valve for the primer / buffer solution output bag is closed. Note that in some embodiments, these or other steps that the system instructs the user to perform may instead be performed automatically by the system, such as by electronically actuating valves.

[0270] Screen 1402R may include a next icon 1422 that may be tapped or clicked by the user to move to the next screen. In some embodiments of screen 1402L, selecting next icon 1422 may cause the system to initiate the remainder of the cell suspension flow process, such as by flowing the cell suspension through the constriction cartridge under pressure. In some embodiments, the next screen may be displayed automatically, and the system may automatically initiate the remainder of the cell suspension flow process once the system detects that cell suspension preparation is complete and / or that system components, such as various valves, are in the correct orientation for the cell suspension to flow through the constriction cartridge and into the correct output bag.

[0271] 14S illustrates screen 1402S, which is displayed on display 1400. Screen 1402S may, in some embodiments, be displayed during all or part of the cell suspension flow process, such as the period during which the cell suspension flows through the constriction cartridge.

[0272] Screen 1402S may include a dynamic pressure indicator 1470 that may display a dynamic indication of the current internal pressure of a system component or components (e.g., preparation vessel, constriction cartridge, etc.) during the cell suspension flow process. Screen 1402S may also include a dynamic temperature indicator 1472 that may display a dynamic indication of the current temperature of one or more system components or contents (e.g., the calculated effective temperature of the cell suspension) during the cell suspension flow process.

[0273] Screen 1402S may include a first dynamic process timer indicator 1474 that may display a dynamic indication of a first current elapsed time for the payload delivery process, the cell suspension flow process, and / or any one or more other sub-processes of the overall payload delivery process. Screen 1402S may include a second dynamic process timer indicator 1476 that may display a dynamic indication of a second current elapsed time for the payload delivery process, the cell suspension flow process, and / or any one or more other sub-processes of the overall payload delivery process. The second current elapsed time may be different from the first current elapsed time. For example, in FIG. 14S, the first dynamic time indicator 1474 indicates the time for the overall process (e.g., starting from priming or cell suspension preparation), while the second dynamic time indicator 1476 indicates the time for the cell suspension flow process (e.g., starting from the cell suspension preparation process or the time the cell suspension begins flowing through the constriction cartridge). In some embodiments, one time indicator may indicate the total time the cell has been in the system, while another time indicator may indicate the total time pressure has been applied to the cell.

[0274] Screen 1402S may include an agitation stop icon 1478 that may be tapped or clicked by the user to stop the system's agitation process, such as by causing the system to send a signal to the shaker plate or other agitation device to stop agitating the cell suspension. In some embodiments, the user may wish to stop the agitation process, for example, when only a small amount of fluid remains in the preparation vessel and continued agitation would risk passing bubbles through the constriction cartridge, or when a large amount of fluid is present and agitation could cause overflow out of the preparation vessel.

[0275] In some embodiments, the system may automatically display the following screen upon detecting that the cell suspension flow process is complete. This detection may be performed according to any one or more of the sensors discussed herein, such as a bubble sensor that monitors the flow, or by monitoring the sample volume (e.g., the volume of fluid in the preparation vessel) during the cell suspension flow process.

[0276] 14T illustrates screen 1402T, which is displayed on display 1400. Screen 1402T may be displayed when the system detects that the cell suspension flow process is complete. For example, when a flow sensor (e.g., flow sensor 912 of system 100) detects that the cell suspension is no longer flowing through one or more tubes of the system, the system may determine that the cell suspension flow process is complete and may display screen 1402T. Screen 1402T, in some embodiments, may prompt the user to confirm that the cell suspension flow process is complete.

[0277] Screen 1402T may include a cell suspension flow completion confirmation icon 1480 that may be tapped or clicked by the user to indicate whether the cell suspension flow process is complete. In some embodiments, indicating that the cell suspension flow process is complete (e.g., by tapping the "Yes" icon) may cause the system to proceed to the next step and display the next screen. In some embodiments, indicating that the cell suspension flow process is not complete may cause the system to continue the cell suspension flow process for a predetermined period of time until the user indicates otherwise or until the system again detects that the cell suspension flow process is complete. In some embodiments, confirming that the cell suspension flow process is complete may cause the system to operate one or more valves or other system components to close the cell suspension flow path.

[0278] 14U illustrates a screen 1402U displayed on the display 1400. Screen 1402U may be a screen displayed to instruct a user to remove all or part of a disposable assembly, such as a preparation vessel assembly, a filter assembly, and / or a gas assembly. In some embodiments, the system may display instructions for disposable assembly removal each time a payload delivery process is performed, such as when the disposable assembly is configured for one-time use only. In some embodiments, the system may read data from one or more sensors to determine whether the disposable assembly has already been removed, and may display the disassembly / removal instructions only if the assembly has not already been removed.

[0279] As shown, screen 1402U may include disposable assembly removal instructions 1422, which may be any graphical and / or textual instructions on how to detach and remove the disposable assembly. In some embodiments, screen 1402U may include disposable assembly removal images (not shown), which may be images or videos illustrating one or more portions of the instructed removal process. In the example of FIG. 14U, the disposable assembly removal instructions instruct the user to seal the tubing to the output bag (which may be done automatically by an electronic valve in some embodiments) and remove the disposable assembly from the system. In some embodiments, the instructions displayed may depend on settings selected by the user in one or more previous screens.

[0280] The screen 1402U may include a next icon 1422 that may be tapped or clicked by the user to move to the next screen. In some embodiments, the system may automatically display the next screen upon detecting that removal of the disposable assembly is complete.

[0281] 14V illustrates screen 1402V, which is displayed on display 1400. Screen 1402V may be a process summary screen that is displayed after the payload delivery process is completed.

[0282] The screen 1402V may include a pressure indicator 1484 that may display an indication of the pressure set point used for the just-completed payload delivery process. Alternatively, or in addition, the pressure indicator 1484 may show one or more actual pressure measurements obtained during the payload delivery process, such as the maximum, minimum, and / or average pressure measured during the process.

[0283] Screen 1402V may include a first static time indicator 1486 that may display an indication of the total elapsed time for the overall payload delivery process. Screen 1402V may include a second static time indicator 1488 that may display an indication of the total elapsed time for the cell suspension flow process (or for any one or more other sub-processes included within the overall payload delivery process). The second total time may differ from the first total elapsed time. For example, in FIG. 14V, first static time indicator 1486 shows the time for the overall process (e.g., starting from priming or cell suspension preparation), while second static time indicator 1488 shows the time for the cell suspension flow process (e.g., starting from the cell suspension preparation process or the time the cell suspension begins flowing through the constriction cartridge). In some embodiments, one time indicator may show the total time the cells have been in the system, while the other time indicator may show the total time pressure has been applied to the cells.

[0284] Screen 1402V may include a temperature control selection indicator 1490 that may indicate whether a temperature control process was selected by the user and / or executed by the system during the process just completed. In some embodiments, screen 1402V may also display information about the temperature control process, such as the elapsed time during the process, the beginning and ending temperatures during the process, the average temperature during the process, and / or a graph depicting the temperature during the process over time.

[0285] Screen 1402V may include an agitation selection indicator 1492 that may indicate whether an agitation process was selected by the user and / or performed by the system during the just-completed process. In some embodiments, screen 1402V may also display information about the agitation process, such as the rate, duration, intensity, and / or an indication of whether and for how long the user stopped the agitation process.

[0286] In some embodiments, the screen 1402V may include a temperature indicator (not shown) that may display an indication of the temperature setpoint used for the just-completed payload delivery process. Alternatively, or in addition, the temperature indicator may show one or more actual temperature measurements (or calculated effective temperatures) obtained during the payload delivery process, such as the maximum temperature, minimum temperature, and / or average temperature measured during the process.

[0287] Screen 1402V may include a back to start page icon 1494 that may be tapped or clicked by a user to signal an input to instruct the system to return to a start screen such as screen 1402B. In some embodiments, a back to start page icon such as icon 1494 may be included on any one or more of the other screens discussed herein with respect to FIGS. 14A-14V.

[0288] In some embodiments, screen 1402V may include a dedicated icon or other user interface object (not shown) for storing or transmitting data detected and / or logged by the system during the just-completed payload delivery process. In some embodiments, the system may be configured to automatically log data related to temperature, pressure, agitation, flow rate, processing time, impedance, light-based sensor data, cell concentration, and / or membrane disruption based on information detected by any one or more sensors during the payload delivery process. In some embodiments, the system may be configured to automatically store and / or transmit the logged information upon completion of the process in response to a user executing an instruction to do so and / or in response to a user tapping or clicking the return to start page icon 1494.

[0289] In some embodiments, any one or more of the inputs made by the user via interface 1400 may be replaced by the user's instructions of a pre-defined routine or recipe that may pre-determine multiple settings (e.g., temperature settings, pressure settings, agitation settings, etc.) and cause the system to perform the payload delivery process according to the pre-defined routine or recipe. Additional intracellular payload delivery systems

[0290] 15-20 illustrate exemplary embodiments of a tabletop laboratory system and associated devices and components for intracellular payload delivery, including a flexible bag usable therein, which systems and devices may be used in conjunction with the methods, techniques, and user interfaces described herein. The systems described in FIGS. 15-20 may share any one or more characteristics in common with the systems described above in FIGS. 1-11, including sharing common components, sharing common characteristics, and / or being usable in all or some of the same methods and / or techniques as described herein. The components, features, and uses described with respect to FIGS. 15-20 may be combined with the components, features, and uses described with respect to FIGS. 1-11. Like the systems described in FIGS. 1-11, the systems described in FIGS. 15-20 may be usable in the methods described with respect to FIGS. 11 and 12 and / or in conjunction with the user interfaces described with respect to FIG. 14.

[0291] 15A-15C illustrate a tabletop system 1500 for delivering a payload to cells, according to some embodiments. Like system 100 described above with respect to FIG. 1 (and other figures referencing system 100), system 1500 may be a tabletop system, such as a piece of laboratory equipment, configured to receive a cell suspension fluid, process the cell suspension fluid, and deliver a payload to cells in the cell suspension. System 1500 and its components / features may share any one or more properties in common with system 100 and / or its individual components / features and / or system 1000 and its individual components / features, and system 1500 may be used in whole or in part in any of the same manners, methods, and / or techniques.

[0292] 15A-15C and described herein, system 1500 may differ from system 100 and / or system 1000 in several respects. That is, system 1500 may have a different physical shape than system 100 defined by its housing, may have a different temperature control system than system 100, and / or may have a different preparation vessel and preparation vessel housing than system 100. With respect to the preparation vessel and preparation vessel housing, system 1500 may utilize a flexible preparation bag inside a rigid preparation housing rather than the rigid preparation vessel of system 100. As described below, using a flexible bag instead of a rigid preparation vessel may improve the cooling functionality of the system because heat transfer to and from the inside of a liquid flexible bag may be more efficient than heat transfer to and from a liquid inside a rigid plastic container.

[0293] As shown in FIG. 15A, the system 1500 may include a base plate 1502, a housing 1503, a hook 1504, an input bag 1506, a preparation vessel housing 1510, an output bag tray area 1511, an output bag 1512, a display 1520, and a constriction cartridge 1524.

[0294] The base plate 1502 may be a platform upon which one or more components of the system 1500 are mounted. For example, the system 1500 may be a tabletop system mounted on the base plate 1502. The base plate 1502 may share any one or more characteristics in common with the platform 102 described above with respect to FIG. 1 . In some embodiments, the base plate 1502 may be separate from one or more other components of the system 1500, while in some embodiments, the base plate 1502 may be integrated (e.g., formed as a single piece) with one or more other components of the system 1500. The base plate 1502 may be made from any suitable material, including metal or plastic. Metal may be preferred to ensure that the base plate 1502 is sufficiently sturdy and heavy to give the system 1500 a low center of gravity.

[0295] The housing 1503 may be any outer housing for the system 1500 and may define the outer shape of the system 1500 and protect the internal components from damage and / or contamination. The housing 1503 may be made from one piece or from two or more separate pieces. In the example shown in FIGS. 15A-15C , the housing 1503 comprises a lower / rear portion and an upper / front portion, and the two portions may be assembled together (e.g., by snapping together, screwing together, or bonding together with an adhesive) to form the housing 1503. The housing 1503 may be made from any suitable material, including metal or plastic. Metal may be preferred to ensure that the system 1500 is sufficiently durable. Plastic may be preferred to ensure that the system 1500 is not excessively heavy and that the center of gravity of the system 1500 is not elevated excessively above the base plate 1502. Materials suitable for use in a clean room environment may be preferred.

[0296] In some embodiments, housing 1503 may share any one or more characteristics in common with the housings of pressure control module 116, temperature control module 118, and / or preparation vessel housing 110. In some embodiments, housing 1503 may contain any one or more of the components of system 100 described above with respect to FIG. 1 , including components located inside or outside of one or more of pressure control module 116, temperature control module 118, and / or preparation vessel housing 110. In some embodiments, system 1500 may provide a more streamlined structure compared to system 100, in which more internal components may be located inside a single housing structure rather than multiple separate housing structures.

[0297] Hook 1504 may be a structure configured to suspend one or more bags, such as bags containing cell suspensions and / or other media. As shown in FIG. 15, hook 1504 is configured to suspend input bag 1506, which may share any one or more features in common with bags 106 and / or 108 described above with respect to FIG. 1. Hook 1504 may likewise share any one or more features in common with hook 104 described above with respect to FIG. 1. As with system 100, the flow path in system 1500 may originate from input bag 1506 and lead through one or more pipes or flexible tubing to other system components, including one or more flexible bags inside the interior of preparation vessel housing 1510 (in system 1500).

[0298] In some embodiments, the hook 1504 may be movable between an extended position and a collapsed position. In the extended position, the hook 1504 may extend upward above the preparation vessel housing 1510 to suspend the bag 1506, while in the collapsed position, the hook 1504 may be located closer to other system components so that the system 1500 occupies less space overall when the hook 1504 is in the collapsed position. In some embodiments, the hook 1504 may move between the extended position and the collapsed position by rotating on a hinge, sliding along a sliding track, or being removed from the system 1500 and reattached to the extended position. In some embodiments, the hook 1504 may be collapsed into the housing 1503 in the collapsed position, or may be parallel and / or flush with one or more walls of the housing 1503 in the collapsed position.

[0299] The preparation vessel housing 1510 may be any structure or component configured to house a preparation vessel. The preparation vessel housing 1510 may share any one or more features in common with the preparation vessel 110 described above with respect to FIG. 1, including being configured to house a preparation vessel containing a cell suspension fluid as it is prepared for passage through a constriction component defining a portion of a flow path configured to create perturbations within a membrane of cells of the cell suspension fluid to facilitate entry of a payload into the cells through the membrane, including by holding the cell suspension while the suspension is cooled (or heated) and agitated as air pressure is applied thereto and / or as the cell suspension is otherwise manipulated or controlled to be forced through the constriction component.

[0300] As described in more detail below with respect to Figures 15-19, the preparation vessel may comprise a flexible bag, such as a flexible plastic bag, instead of and / or in addition to a rigid vessel, such as vessel 600 described above with respect to Figure 6A and / or system 100. Preparation vessel housing 1510 may therefore differ from preparation vessel 100 in that preparation vessel housing 1510 may be configured to store a preparation vessel in the form of a flexible bag rather than in the form of a rigid vessel. Preparation vessel housing 1510 may be configured to store a preparation vessel in the form of a flexible bag by having an internal cavity within the shape of the filled flexible bag. In this way, when the flexible bag is filled with fluid and / or pressurized gas, it may be pressed against the interior wall of preparation vessel housing 1510 such that preparation vessel housing 1510 may create a cavity enclosed by the bag.

[0301] One advantage of using a preparation vessel in the form of a flexible bag is that there may be increased efficiency in heat transfer to and / or from the fluid inside the bag while inside the preparation vessel housing 1510. Because the walls of a flexible bag (e.g., a plastic flexible bag) may be significantly thinner than the walls of a rigid container, and because the walls of a flexible bag may deform to increase the surface area of ​​contact between the bag and the interior walls of the preparation vessel housing 1510, heat transfer to and from the fluid inside the bag may be more efficient and faster than with thicker walls and less surface area contact.

[0302] To facilitate this efficient heat transfer, the preparation vessel housing 1510 may comprise a temperature control system. In some embodiments, the temperature control system of the preparation vessel housing 1510 may share any one or more characteristics in common with any of the components of and / or associated with the temperature control module 118 discussed above with respect to FIG. 1. In some embodiments, the temperature control system of the preparation vessel housing 1510 may comprise any one or more components configured to heat and / or cool a fluid inside the preparation vessel inside the preparation vessel housing 1510, such as one or more forced air heaters, one or more forced air coolers, one or more thermoelectric cooling devices (e.g., Peltier coolers), one or more resistive heating devices, one or more liquid heating devices, one or more liquid cooling devices, or the like.

[0303] In some embodiments, one or more thermoelectric cooling devices (e.g., cooling plates) may be integrated into the preparation vessel housing 1510 such that they may form part of one or more interior walls of the preparation vessel housing 1510. A flexible bag or other preparation vessel inside the preparation vessel housing 1510 may then contact the thermoelectric cooling device (e.g., cooling plate), thereby drawing heat from the fluid inside the vessel and cooling the fluid therein. In some embodiments, the flexible bag (as described in more detail below) may have a front side and a back side that form a large flat area over which the bag may contact the thermoelectric cooling plate. In some embodiments, the preparation vessel housing 1510 may comprise a first thermoelectric cooling plate configured to contact the front side of the flexible bag and a second thermoelectric cooling plate positioned opposite the first plate inside the thermoelectric cooling plate configured to contact the back side of the flexible bag.

[0304] The preparation vessel housing 1510 may include one or more openable doors. The one or more doors may be movable between an open position and a closed position, for example, by being movably mounted on a hinge. Opening one or more doors of the preparation vessel housing 1510 may allow for removal and / or replacement of a preparation vessel (e.g., a flexible bag) (and / or any other component) located inside the preparation vessel housing 1510. In some embodiments, one or more thermoelectric cooling devices (e.g., cooling plates) may be integrated into the doors of the preparation vessel housing 1510 such that closing the doors may press the one or more thermoelectric cooling devices against the preparation vessels positioned inside the preparation vessel housing 1510.

[0305] In some embodiments, system 1500 may be configured such that positive pressure in a container inside a flexible bag inside the preparation vessel housing 1510 ensures that the outer walls of the flexible bag are pressed into contact with the inner walls of the preparation vessel housing 1510, thereby allowing fluid to be expelled from the flexible bag and still be used to ensure an effective surface area for temperature control. To provide positive pressure inside the flexible bag, gas (such as sterilizing gas or pressurized air) may be pumped inside the flexible bag at a pressure sufficient to cause the bag to expand and / or press against the inner walls of the preparation vessel housing 1510. In some embodiments, the pressure sufficient to cause the bag to expand and / or press against the inner walls of the preparation vessel housing 1510 may be greater than 2 psi, 5 psi, 10 psi, 25 psi, 40 psi, 80 psi, 100 psi, 120 psi, 140 psi, or 160 psi. In some embodiments, the pressure sufficient to cause the bag to expand and / or press against the inner wall of the preparation vessel housing 1510 may be less than 2 psi, 5 psi, 10 psi, 25 psi, 40 psi, 80 psi, 100 psi, 120 psi, 140 psi, or 160 psi.

[0306] In some embodiments, one or more thermoelectric cooling devices, such as one or more cooling plates, may be positioned inside the preparation vessel housing 1510 such that they can contact the fluid inside the flexible bag inside the preparation vessel housing 1510 even when the fluid level is low. For example, the cooling plates may be positioned at or near the bottom of the preparation vessel housing 1510 such that even when the fluid level in the vessel is below 50%, below 25%, below 10%, or below 5%, all or a portion of the surface area of ​​the one or more cooling plates can still contact all or a portion of the portion of the vessel that is in contact with the fluid remaining in the vessel.

[0307] In some embodiments where the one or more thermoelectric cooling devices comprise one or more cooling plates, the plates may be surrounded by insulation on the back and / or to the sides, hi some embodiments, the insulation may be 3D printed to fit snugly over the plates.

[0308] In some embodiments, one or more individual thermoelectric cooling plates used within the preparation vessel housing 1510 may have a temperature of 10,000 mm 2 , 12,100mm 2 , 14,400mm 2 , 16,900mm 2 , or 19,600 mm 2 In some embodiments, one or more individual thermoelectric cooling plates used within the preparation vessel housing 1510 may have a functional surface area of ​​greater than 10,000 mm 2 , 12,100mm 2 , 14,400mm 2 , 16,900mm 2 , or 19,600 mm 2 It may have a functional surface area of ​​less than

[0309] In some embodiments, the one or more temperature control devices used in the preparation vessel housing 1510 may comprise a temperature probe, such as a hot-side temperature probe, configured to measure the coolant temperature entering a radiator for fan control of the device. In some embodiments, the one or more temperature control devices used in the preparation vessel housing 1510 may comprise a cold-side temperature probe, such as one embedded in a cooling plate, configured to take a temperature reading of the cooling plate.

[0310] In some embodiments, one or more temperature control devices used within the preparation vessel housing 1510 may be configured to heat and / or cool the fluid inside the vessel inside the preparation vessel housing 1510 according to any one or more of the temperature ranges and / or time ranges discussed with respect to block 1324 above. In some embodiments, the system 1500 may be configured to cool the fluid inside the preparation vessel housing 1510 from above 20 degrees Celsius, 22 degrees Celsius, 24 degrees Celsius, 30 degrees Celsius, 34 degrees Celsius, 36 degrees Celsius, or 38 degrees Celsius to below 8 degrees Celsius, 6 degrees Celsius, 5 degrees Celsius, 4 degrees Celsius, 2 degrees Celsius, or 1 degree Celsius. In some embodiments, the system 1500 may be configured to cool the fluid inside the preparation vessel housing 1510 over one or more of these ranges in a period of less than 1 hour, 45 minutes, 30 minutes, 15 minutes, 10 minutes, or 5 minutes.

[0311] Once the fluid inside the preparation vessel housing 1510 is prepared for passage out of the preparation vessel housing 1510 (e.g., toward and through the constriction cartridge), the fluid may be forced out of an outlet of a flexible bag in the preparation vessel housing 1510, through one or more tubes or pipes, and toward a downstream component, such as a constriction cartridge. As discussed above with respect to system 100, fluid may be forced out of the preparation vessel of system 1500 by pressurizing a gas inside the preparation vessel. Because the preparation vessel in system 1500 may be a flexible bag, the preparation vessel housing 1510 may provide a rigid structure (e.g., a shell) that prevents the flexible bag preparation vessel from bursting or otherwise breaking when the bag is pressurized. In some embodiments, gas pressures the same or similar to those discussed above with respect to forcing fluid out of vessel 600 in system 100 may be used.

[0312] In some embodiments, the preparation vessel housing 1510 may comprise (and / or comprises) one or more of: (a) a flow sensor (e.g., a foam sensor) upstream of the preparation vessel that may be used to monitor when all liquid has entered the preparation vessel from an upstream component such as an input bag, (b) a flow sensor (e.g., a foam sensor) downstream of the preparation vessel that may be used to monitor when all liquid has exited the preparation vessel and flowed toward a downstream component such as a constriction cartridge, and (c) one or more level sensors on and / or in the preparation vessel housing 1510 configured to sense the level of fluid in the preparation vessel while it is within the preparation vessel housing 1510. In some embodiments, one or more of these components may be used to sense the flow of fluid into and / or from the preparation vessel housing 1510 and / or to sense the level of fluid in the preparation vessel inside the preparation vessel housing 1510. Sensing these characteristics may, in some embodiments, be used to implement one or more automated aliquot functionality, such as when it is desired to process a portion of the fluid in the input bag and / or preparation vessel in one manner at one time and process the remainder of the fluid in another manner and at another time.

[0313] Constriction cartridge 1524 may be any structure configured to contain or store a component having a narrowing component, such as a narrowing filter (containing one or more narrowed microfluidic pores) or a narrowing microfluidic chip (containing one or more narrowed microfluidic channels). In some embodiments, constriction cartridge 1524 may share any one or more characteristics in common with any one or more constriction cartridges discussed above with reference to FIGS. 1, 5A, and / or 5B, constriction cartridge 700 discussed above with reference to FIGS. 7A-7C, and / or constriction cartridge 800 discussed above with reference to FIGS. 8A-8D. Similar to the other constriction cartridges discussed herein, constriction cartridge 1524 may receive a flow of prepared cell suspension from a preparation vessel downstream and flow the cell suspension through one or more narrowing components, such as a narrowing filter or any component containing a narrowing channel, passage, or other small opening, such as a narrowing tip, contained inside constriction cartridge 1524. After passing through the narrowing components, the suspension may flow out of the constriction cartridge toward one or more downstream components of system 1500, such as an output bag.

[0314] Output bag 1512 may be fluidly connected to constriction cartridge 1524 and configured to receive a flow of fluid, such as a cell suspension fluid, from constriction cartridge 1524. Output bag 1512 may share any one or more characteristics in common with output bags 112 and 114, as discussed above with reference to system 100 and FIG.

[0315] Output bag tray area 1511 may be a platform, flat space, or other area included within system 1500 configured to allow output bags, such as output bag 1512, to rest on the area during use of system 1500, including before, during, and / or after the output bags are filled. In some embodiments, output bag tray area 1511 may share any one or more characteristics in common with output bag tray 111, as discussed above with reference to system 100 and FIG. 1. As shown in FIG. 15A , the output bag tray area may be integrally formed as part of housing 1503. Alternatively, or in addition, in some embodiments, the output bag tray area may be formed as part of a base plate, such as base plate 1502.

[0316] In some embodiments, the output bag tray area 1511 may comprise a temperature control system. In some embodiments, the temperature control system of the output bag tray area 1511 may share any one or more characteristics in common with any of the components associated with and / or the temperature control module 118 discussed above with respect to FIG. 1 and / or the temperature control system of the preparation vessel housing 1510 discussed above with respect to FIG. 15. In some embodiments, the temperature control system of the output bag tray area 1511 may comprise any one or more components configured to heat and / or cool a fluid inside the output bag 1512 while the output bag 1512 rests on the output bag tray area 1511, such as one or more forced air heaters, one or more forced air coolers, one or more thermoelectric cooling devices (e.g., Peltier coolers), one or more resistive heating devices, one or more liquid heating devices, one or more liquid cooling devices, or the like. In some embodiments, the temperature control system for output bag tray area 1511 may comprise a heating plate that may be disposed on and / or integrated into a flat surface of output bag tray area 1511. The heating plate or other temperature control component integrated into or associated with output bag tray area 1511 may be used in some embodiments to heat the output sample to temperatures above room temperature, 35 degrees Celsius, 37 degrees Celsius, 39 degrees Celsius, or above, and / or any temperature above the temperature at which cells are processed by system 1500.

[0317] In some embodiments, output bag tray area 1511 may include a cover configured to cover output bags 1512, thereby shielding it from physical contact, airborne contaminants, and / or light. In some embodiments, the cover for the output bag tray may be a removable and / or replaceable lid, such as a lid mounted on a hinge and / or sliding track.

[0318] In some embodiments, the output bag tray area 1511 may include, and / or the system 1500 may include, one or more agitation devices configured to agitate the output bags 1512 while they rest on the output bag tray area 1511. In some embodiments, the one or more agitation devices may share any one or more characteristics in common with other agitation devices discussed herein, including, but not limited to, one or more shaker plates, vibration devices, kneading devices, sonic agitation devices, peristaltic pump devices, gas / diaphragm devices, or other mechanisms configured to cause shaking / vibration of the output bags 1512 and / or agitation / circulation of the fluid therein.

[0319] Display 1520 may be any display, such as a touchscreen display, configured to display one or more graphical elements and / or a graphical user interface related to the operation of system 1500. In some embodiments, display 1520 may share any one or more characteristics in common with display 120 discussed above with reference to FIG. 1. Display 1520, in some embodiments, may be used to control any one or more functions of system 1500, including by displaying and / or receiving user input via the user interface described with respect to FIG. 14, as discussed above.

[0320] 15B, system 1500 may include a door 1522, which may be any door, access hatch, or equivalent, movable between an open position and a closed position and configurable to allow access to one or more components located inside housing 1503. In the example of FIG. 15B, door 1522 is located on the side and / or back of housing 1503.

[0321] 16 illustrates a schematic diagram of a tabletop system 1600 for delivering a payload to a cell, according to some embodiments. In some embodiments, system 1600 may share some or all characteristics in common with system 100 described above with respect to FIG. 1 and / or with system 1000 described above with respect to FIG. 10.

[0322] Rather than depicting the physical configuration of the various components of a system for delivering a payload to cells, FIG. 16 primarily depicts diagrammatically the flow paths and associated components for fluids (e.g., cell suspension, buffer fluid) traveling through the system and for pressurized gas traveling through the system. That is, FIG. 16 depicts various components through which cell suspension and / or buffer fluid may flow while being processed by the system, and various components through which gas (e.g., pressurized gas) may flow as it is being passed through the system. FIG. 16 shows components fluidly connected to one another via representations of tubing, piping, or the like connecting the representations of the components. Unless otherwise noted, any suitable tubing or piping may be used to fluidly connect the various components, such as flexible plastic tubing, rigid plastic tubing, PVC tubing, metal tubing, or the like.

[0323] System 1600 may include an input bag 1602, which may share some or all characteristics in common with input bag 1506 and / or with any other input bag described herein. In some embodiments, the input bag may have a 1 L volume, a 2 L volume, or a 3 L volume. In some embodiments, the flow path of the liquid flowing through system 1600 may originate (or part of) from input bag 1602.

[0324] System 1600 may include a first flow sensor 1616, which may share any one or more characteristics in common with flow sensor 912 discussed above with reference to Figures 9C and 9D and / or with flow sensor 1062 discussed above with reference to Figure 10. Flow sensor 1616 may be configured to detect when fluid flow has stopped, thereby enabling system 1600 to determine when an input bag is empty and / or when all of the fluid sample has flowed from an input bag (e.g., bag 1602) into a preparation vessel (e.g., bag 1606).

[0325] The system 1600 may include an upper automated tubing occluder 1604, which may be any valve, clamp, cap, or equivalent used to control the flow of fluid from the input bag 1602. The automated tubing occluder 1604 may be controlled to enable or disable fluid flow automatically (e.g., electronically) according to instructions executed by a processor of the system 1500, so that manual actuation by a user is not required. In some embodiments, a manual tubing occluder operated by hand may be used.

[0326] System 1600 may include a reservoir bag 1606 that may share some or all characteristics in common with a preparation vessel in the form of a flexible bag discussed above with reference to system 1500 in Figure 15, with any of the flexible bags described below with reference to Figures 17, 18, and / or 19, and / or with any flexible bag serving as a preparation vessel described herein. As indicated by the dotted line surrounding reservoir bag 1606, reservoir bag 1606, and several other components, reservoir bag 1606 may be located inside a preparation vessel housing, such as preparation vessel housing 1510.

[0327] System 1600 may include gas pressure inlet 1608, which may be any opening or inlet configured to allow gas to flow into reservoir bag 1606. As discussed above, a preparation vessel in the form of a flexible bag may be filled with pressurized gas (e.g., sterilizing gas and / or air) to expand the bag and contact the interior walls of the preparation vessel housing. Gas pressure inlet 1608 may be used to direct a flow of gas into reservoir bag 1606 for said pressurization and subsequent flow through the constriction cartridge.

[0328] System 1600 may include a temperature control system 1610 that may share some or all characteristics in common with the temperature control system of preparation vessel housing 1510 described above with respect to Figure 15 and / or with any other temperature control system described herein. In some embodiments, temperature control system 1610 may be a cooling system, such as a thermoelectric cooling system, configured to cool the cell suspension fluid while the fluid is inside reservoir bag 1606.

[0329] System 1600 may include a peristaltic pump 1612 that may be configured to drive fluid flow through a circulation loop fluidly connected to reservoir bag 1606. By pumping fluid from reservoir bag 1606, through the circulation loop, and back into reservoir bag 1606, peristaltic pump 1612 may circulate fluid inside reservoir bag 1606 during fluid preparation, such as during a cooling process. Fluid circulation may promote cell mixing and / or prevent cells from settling out of suspension during the cooling process, other preparation processes, and / or system rest. This may lead to more consistent performance and more uniform cell concentration as the cell suspension fluid flows through constriction cartridge 1622.

[0330] System 1600 may include a second flow sensor 1620, which may share some or all characteristics in common with first flow sensor 1616 described above and / or with any one or more other flow sensors described herein. Flow sensor 1620 may be configured to detect when fluid flow has stopped, thereby enabling system 1600 to determine when the preparation vessel / bag has emptied and / or when all fluid has flowed into downstream components, such as constriction cartridge 1622. In some embodiments, system 1600 may stop a system step (and / or other system process) when sensor 1620 detects that the flow of liquid from the preparation vessel has slowed or stopped, for example, by ceasing pressurization of the preparation vessel and / or by closing one or more valves.

[0331] System 1600 may include filter 1618, which may be a filter configured to remove or break up cell aggregates and / or other debris from the prepared cell suspension while allowing other fluids and the single cell suspension to pass through filter 1618 toward constriction cartridge 1622. In some embodiments, filter 1618 may include a filter element housed inside an external housing, the filter element and external housing configured to withstand high pressures of fluid being forced through filter 1618, including maximum pressures of greater than or equal to 40 PSI, greater than or equal to 80 PSI, greater than or equal to 120 PSI, or greater than or equal to 160 PSI. In some embodiments, filter 1618 may be configured to withstand maximum pressures of greater than or equal to 40 PSI, greater than or equal to 80 PSI, greater than or equal to 120 PSI, or less than 160 PSI. In some embodiments, the filter element may be removable from the external housing so that a used filter element can be removed and replaced without having to replace the external housing as well. In some embodiments, this filtering process enabled by filter 1618 may reduce clogging of constrictions in the narrowing tip and / or narrowing filter. In some embodiments, this filtering process may reduce the occurrence of clogging by 10 times or more, 50 times or more, or 100 times or more. The reduced clogging may, in some embodiments, improve system throughput by (a) preventing throughput from being reduced as system components become partially clogged, and (b) reducing or preventing the need to stop or halt system processes to repair or replace components that become substantially or completely clogged.

[0332] System 1600 may include a third flow sensor 1621, which may share some or all characteristics in common with first flow sensor 1616 and / or second flow sensor 1620 described above and / or with any one or more other flow sensors described herein. Flow sensor 1621 may be configured to detect when fluid flow has stopped, thereby allowing system 1600 to determine when the tubing has emptied of fluid and / or when all fluid has flowed through any upstream components, such as filter 1618. In some embodiments, flow sensor 1621 may be used when flushing system 1600, such as to ensure that no fluid remains in the system tubing after a process is completed.

[0333] System 1600 may include constriction cartridge 1622, which may share some or all characteristics in common with constriction cartridge 1524 described above with reference to FIG. 15, any one or more of the constriction cartridges discussed above with reference to FIGS. 1, 5A, and / or 5B, constriction cartridge 700 discussed above with reference to FIGS. 7A-7C, and / or constriction cartridge 800 discussed above with reference to FIGS. 8A-8D. As shown in FIG. 16, fluid (e.g., cell suspension fluid) may flow from filter 1618 toward and into constriction cartridge 1622, where the fluid may be forced through one or more constrictions. The fluid may then flow out of constriction cartridge 1622 toward and into output bag 1630.

[0334] The system 1600 may include a lower automated tubing occluder 1624, which may share some or all characteristics in common with the upper automated tubing occluder 1604 discussed above. The lower automated tubing occluder 1624 may be any valve, clamp, cap, or equivalent used to control flow to the fluid cartridge 1622 and output bag 1626. The automated tubing occluder 1624 may be controlled to enable or disable fluid flow automatically (e.g., electronically) according to instructions executed by a processor of the system 1500, so that manual actuation by a user is not required. In some embodiments, a manual tubing occluder operated by hand may be used.

[0335] System 1600 may include an output bag 1626 that may share some or all characteristics in common with output bag 1512 discussed above with reference to FIG. 15 and / or with any one or more other output bags described herein. In some embodiments, the output bag may have a volume of 2 L, a volume of 3 L, or a volume of 4 L. In some embodiments, the flow path of the liquid flowing through system 1600 may terminate (or be partially terminated) in output bag 1626.

[0336] The system 1600 may include a leak-containing tray 1628 that may share some or all characteristics in common with the output bag tray area 1511 described above with reference to Figure 15 and / or with the output bag tray 111 described above with reference to Figure 1. In some embodiments, the leak-containing tray 1628 may have one or more raised edges, one or more recessed portions, one or more walls, one or more absorbent elements, and / or one or more other physical features configured to contain leak-through from the output bag 1626 or associated tubing.

[0337] System 1600 may include an output bag cover 1630 that may share some or all characteristics in common with the cover associated with output bag 1512 described above with reference to FIG. 15. In some embodiments, the bag cover may shield the output bag from physical contact, airborne contaminants, and / or light. In some embodiments, the cover for the output bag may be a removable and / or replaceable lid, such as a lid mounted on a hinge and / or sliding track.

[0338] System 1600 may include a temperature control system 1632 that may share some or all characteristics in common with the temperature control system for output bag tray area 1511 described above with reference to FIG. 15 and / or with any other temperature control system described herein. In some embodiments, temperature control system 1632 may be integrated into or disposed on or within spillage-containing tray 1628 and may be configured to control (e.g., heat) the temperature of output bag 1626 after the treated suspension fluid flows into output bag 1626.

[0339] In some embodiments, one or more components shown in FIG. 16 may together form all or part of a disposable assembly. For example, the components may be configured for one-time use such that they can be used to perform a payload delivery process once and then discarded. That is, a cell suspension may be flowed through the flow paths of system 1600 once, and then some or all of the elements of system 1600 may be replaced before another payload delivery process is performed. In some embodiments, the components of the disposable assembly may be constructed from materials that are suitable for gamma sterilization, such as for use in a sterile environment. In some alternative embodiments, the components of the disposable assembly may be constructed from materials that are suitable for sterilization by other methods, such as autoclaving or ethylene oxide sterilization. In some embodiments, the components of the disposable assembly may be packaged and / or shipped together, such as packaged and / or shipped in a sealed sterile container. In some embodiments, components of the disposable assembly may be configured to be attachable to other components of a system for intracellular payload delivery in a manner suitable for implementation in a sterile environment, such as by hand attachment without the use of tools and / or by using a sterile connector mechanism. In the example of FIG. 16, the disposable assembly may include at least one or more of reservoir bag 1606, filter 1618, and / or cartridge 1622, and associated tubing. In some embodiments, one or more sensors, such as one or more of flow sensors 1616, 1620, and 1621, may also be part of the disposable assembly.

[0340] 17 illustrates a flexible bag 1700 for holding a cell suspension fluid as it is prepared for passage through a constriction component of a tabletop system for delivering a payload to cells, according to some embodiments. Flexible bag 1700 may be configured for use as a preparation vessel within a system for delivering a payload to cells, and in some embodiments may be used in such systems, such as systems 100, 1000, 1500, and / or 1600, as discussed above with reference to FIGS. 1, 10, 15, and 16, respectively. In some embodiments, flexible bag 1700 may share any one or more characteristics in common with the flexible bag described above with reference to preparation vessel housing 1510 in FIG. 15 and / or the flexible bag in the form of reservoir bag 1606 described above with reference to FIG. 16, the preparation vessel, or the like, as described elsewhere herein. In some embodiments, flexible bag 1700 may be used in the same or similar manner as any one or more of those bags otherwise described herein.

[0341] In some embodiments, flexible bag 1700 (and / or any other bag disclosed herein) may be made from PVC, silicone, thermoplastic elastomer (TPE), or any other suitable material. In some embodiments, flexible bag 1700 may be flexible (e.g., bendable) and / or elastic (e.g., stretchable). In some embodiments, flexible bag 1700 may be flexible but not elastic. In some embodiments, bags used in the systems described herein may be elastic but not flexible. In some embodiments, bags used in the systems described herein may be wholly or partially flexible and / or elastic, and / or wholly or partially inflexible and / or inelastic. For example, in some embodiments, the bag may be movable between a flattened configuration and an expanded configuration without stretching and / or one or more portions of bag flex. In some embodiments, the flexible bag 1700 may have one or more dimensions configured to be larger than the preparation vessel housing into which it is inserted so that the bag can be pressurized and expanded to touch the interior walls of the preparation vessel housing without reaching full tension and / or having to stretch.

[0342] Bag 1700 may have a bag wall thickness selected according to requirements for bag strength and flexibility. In some embodiments, one or more physical properties or dimensions of bag 1700 may be selected such that bag 1700 will not fail (e.g., burst) during use in one or more of the systems described herein.

[0343] In some embodiments, the bag 1700 may have a bag wall thickness greater than 0.2 mm, 0.3 mm, 0.4 mm, or 0.5 mm. In some embodiments, the bag 1700 may have a bag wall thickness less than 0.2 mm, 0.3 mm, 0.4 mm, or 0.5 mm.

[0344] In some embodiments, the bag 1700 may have an edge weld thickness greater than 0.4 mm, 0.5 mm, 0.6 mm, or 0.7 mm. In some embodiments, the bag 1700 may have an edge weld thickness less than 0.4 mm, 0.5 mm, 0.6 mm, or 0.7 mm.

[0345] In some embodiments, the bag 1700 may have an edge weld width greater than 2 mm, 3 mm, 4 mm, 5 mm, or 6 mm. In some embodiments, the bag 1700 may have an edge weld width less than 2 mm, 3 mm, 4 mm, 5 mm, or 6 mm.

[0346] In some embodiments, bag 1700 may include one or more welds around the tubing of bag 1700. In some embodiments, the tubing of bag 1700 may have an outer diameter greater than 5 mm, 6 mm, 7 mm, 8 mm, or 9 mm. In some embodiments, the tubing of bag 1700 may have an outer diameter less than 5 mm, 6 mm, 7 mm, 8 mm, or 9 mm.

[0347] In some embodiments, the bag 1700 may include one or more drilled holes (e.g., for mounting). In some embodiments, one or more of the drilled holes may be spaced apart from the weld by a distance greater than 5 mm, 6 mm, 7 mm, 8 mm, or 9 mm. In some embodiments, one or more of the drilled holes may be spaced apart from the weld by a distance less than 5 mm, 6 mm, 7 mm, 8 mm, or 9 mm.

[0348] In some embodiments, bag 1700 may be configured to be capable of withstanding application of high internal pressure while located inside a preparation vessel housing as discussed herein, so as to urge fluid inside fluid bag 1700 to exit bag 1700 and flow toward and into the constriction cartridge. In some embodiments, bag 1700 may be configured to be capable of withstanding application of pressures greater than 130 psi, 140 psi, 150 psi, 160 psi, or 170 psi. In some embodiments, bag 1700 may be configured to be capable of withstanding application of pressures less than 130 psi, 140 psi, 150 psi, 160 psi, or 170 psi.

[0349] In some embodiments, the bag 1700 may have a volume greater than 1 L, 2 L, 3 L, 4 L, 5 L, or 10 L. In some embodiments, the bag 1700 may have a volume less than 1 L, 2 L, 3 L, 4 L, 5 L, or 10 L.

[0350] In some embodiments, the bag 1700 may have a generally flat shape with a flat front side and a flat back side. In some embodiments, the bag 1700 may have a tapered / sloped bottom to promote the flow of liquid toward and out of the outlet (e.g., as discussed below).

[0351] 17, flexible bag 1700 may include a product inlet 1702, an air inlet 1704, a product outlet 1706, and a circulation loop 1708. Bag 1700 may also be provided with a flow sensor 1716.

[0352] Product inlet 1702 may be any fluid inlet configured to allow fluid to flow into bag 1700. Product inlet 1702 may be configured to allow the flow of liquid, such as cell suspension fluid and / or buffer liquid, into bag 1700 during one or more cell treatment processes as described herein. For example, during a buffer process, buffer liquid may flow into bag 1700 via product inlet 1702, and during a cell treatment preparation process, cell suspension fluid may flow into bag 1700 via product inlet 1702. Product inlet 1702 may be located at or near the top of bag 1700 such that liquid product (e.g., cell suspension and / or buffer liquid) entering bag 1700 may enter bag 1700 from the top side and fall to the bottom of the bag under gravity. In some embodiments, the product inlet 1702 may share any one or more characteristics in common with one or more of the vessel inlets 604 described above with reference to FIG. 6.

[0353] Gas inlet 1704 may be any fluid inlet configured to flow into bag 1700. Gas inlet 1704 may be configured to allow a flow of gas, such as a pressurized gas (e.g., sterilizing gas, pressurized air, etc.), to flow into bag 1700 during one or more pressurization processes as described herein. For example, during cooling of the fluid inside bag 1700 while bag 1700 is inside the preparation vessel housing, gas may be flowed into bag 1700 through gas inlet 1704 to promote effective cooling and to force the cell suspension fluid out product outlet 1706, thereby expanding bag 1700 and contacting the interior walls of the preparation vessel housing. Gas inlet 1704 may be located at or near the top of bag 1700 so that gas entering bag 1700 does not bubble through liquid resting in the bottom of the bag. In some embodiments, the gas inlet 1704 may share any one or more characteristics in common with one or more of the vessel inlets 604 described above with reference to FIG. 6.

[0354] Product outlet 1706 may be any fluid outlet configured to allow fluid to flow out of bag 1700. Product outlet 1706 may be configured to allow the flow of liquid, such as cell suspension fluid and / or buffer liquid, out of bag 1700 during one or more cell treatment processes as described herein. For example, during a buffer process, buffer liquid may flow out of bag 1700 via product outlet 1706, and during a cell treatment preparation process, cell suspension fluid may flow out of bag 1700 via product outlet 1706. Product outlet 1706 may be located at or near the bottom of bag 1700 so that liquid product (e.g., cell suspension fluid and / or buffer liquid) exiting bag 1700 can be forced out of outlet 1706 under gravity and / or the force of pressurized gas presses bag 1700 downward. In some embodiments, the product outlet 1706 may share any one or more characteristics in common with one or more of the vessel outlets 606 described above with reference to FIG. 6.

[0355] Circulation loop 1708 may be any one or more components of bag 1700 configured to circulate and / or recirculate fluid within bag 1700 through a fluid path so as to mix the fluid, create turbulence, or otherwise physically agitate the fluid. In the example of FIG. 17A , circulation loop 1708 comprises a tubing loop forming a flow path that exits the main body of bag 1700 at circulation outlet 1710 and returns into the main body of bag 1700 at circulation inlet 1712. Openings 1710 and 1712 may share any one or more characteristics in common with other outlets and inlets (e.g., tubing or plumbing outlets or inlets) described herein. The flow path of circulation loop 1708 may comprise one or more of plumbing or tubing.

[0356] 17, the circulation outlets 1710 are each located near the bottom of the bag 1700 so that liquid can be drawn from the bag 1700 from below the fill level even when the bag is at a very low fill level (e.g., less than 10% full, less than 5% full, or less than 2.5% full). Similarly, locating the circulation inlets 1712 near the bottom of the bag 1700 may allow liquid to be recirculated into the main body of the bag 1700 below the same or similar low fill line, such that the liquid may re-enter the bag 1700 below the surface of the liquid in the main body of the bag. In some alternative embodiments, one or both of the circulation outlets 1710 and the circulation inlets 1712 may be located anywhere on the bag 1700. For example, in some alternative embodiments, the circulation outlet 1710 may be located near the bottom of the bag 1700, while the circulation inlet 1712 may be located near the top of the bag 1700, so that the recirculation loop can cause fluid to flow from near the bottom of the bag 1700 to near the top of the bag 1700 (e.g., causing liquid to descend under gravity back into the main body of the bag 1700).

[0357] Flow through a circulation loop, such as circulation loop 1708, may be driven by pump 1714, which may be any pump configured to cause fluid flow through loop 1708, and in some embodiments, may be a peristaltic pump. In some embodiments, the flow rate through circulation loop 1708 may be determined and / or set according to a sample volume (e.g., the volume of fluid and / or liquid inside bag 1700). In some embodiments, the circulation rate may be determined automatically by the system into which bag 1700 is placed, such as by being determined according to data read from one or more sensors in the system (e.g., data indicating the fluid volume inside bag 1700), while in some embodiments, a user of the system may enter a user input that sets the desired circulation rate. In some embodiments, the circulation rate of bag 1700 may be greater than 100 mL / min, 200 mL / min, 300 mL / min, 400 mL / min, 500 mL / min, or 600 mL / min. In some embodiments, the circulation rate of bag 1700 may be less than 100 mL / min, 200 mL / min, 300 mL / min, 400 mL / min, 500 mL / min, or 600 mL / min. In some embodiments, the circulation loop of a bag, such as bag 1700 or any other bag or preparation vessel disclosed herein, may be configured to circulate more than 10%, 20%, 25%, 30%, 40%, 50%, or 99% of the volume of liquid in the bag during the preparation process. In some embodiments, the circulation loop of a bag, such as bag 1700 or any other bag or preparation vessel disclosed herein, may be configured to circulate less than 10%, 20%, 25%, 30%, 40%, 50%, or 99% of the volume of liquid in the bag during the preparation process.

[0358] Flow sensor 1716 may be provided in conjunction with bag 1700 and may be any sensor configured to detect flow, lack of flow, and / or flow rate associated with bag 1700. As shown in FIG. 17 , flow sensor 1716 may be configured to detect the flow and / or flow rate of fluid flowing from bag 1700 out through product outlet 1706. In some embodiments, flow sensor 1716 may be configured to detect when liquid flows out of outlet 1706 and to detect when gas (e.g., bubbles) is present or absent within the flow of liquid flowing out of outlet 1706. Flow sensor 1716 may share any one or more characteristics in common with any one or more other flow sensors discussed herein, including flow sensor 1616 discussed above with respect to FIG. 16 . The flow sensor 1716 may be configured to detect when the fluid flow has stopped, thereby enabling the system 1600 to determine when the bag 1700 is empty or nearly empty of liquid and when the buffer process is complete.

[0359] 18 illustrates a flexible bag 1800 for holding a cell suspension fluid as it is prepared for passage through a constriction component of a tabletop system to deliver a payload to cells, according to some embodiments. As shown in FIG. 18 , flexible bag 1800 may include a product inlet 1802, an air inlet 1804, a product outlet 1806, a circulation loop 1808, a pump 1814, and a flow sensor 1816. Bag 1800 may also be provided in conjunction with flow sensor 1816. In some embodiments, bag 1800 may share any one or more characteristics in common with bag 1700 described above with reference to FIG. 17 and / or any other flexible bag described herein.

[0360] In some embodiments, flexible bag 1800 may differ from flexible bag 1700 in that the circulation loop of bag 1800 may be integrated into the product outlet of bag 1800. That is, circulation loop 1708 has a dedicated circulation inlet 1710 and circulation outlet 1712, while circulation loop 1808 has a dedicated circulation inlet 1810 but does not have a dedicated circulation outlet. Rather, the outlet for circulation loop 1808 is product outlet 1806, which serves both as a return path for fluid circulation within circulation loop 1808 and as an output path for product exiting bag 1800. This is accomplished by joining the flow path for circulation loop 1808 into the flow path of outlet 1806 at a T-junction or Y-junction. As further shown in FIG. 18, a flow sensor 1816 may be provided along the portion of the tubing / plumbing where the loop 1808 joins the flow path of the outlet 1806 downstream from the pump 1814 and upstream of the join.

[0361] In some embodiments, bag 1700 may be referred to as a "five-port" bag due to the five different inlet / outlet ports on the bag, while bag 1800 may be referred to as a "four-port" bag due to the four different inlet / outlet ports on the bag. As previously mentioned, the four-port bag may have one less port than the five-port bag due to the integration of the circulation loop and product outlet.

[0362] 19A-19D illustrate a flexible bag 1900 for holding a cell suspension fluid during the performance of four different functions of a tabletop system for delivering a payload to cells, according to some embodiments. The flexible bag 1900 may include a product inlet 1902, an air inlet 1904, a product outlet 1906, a circulation loop 1908, a pump 1914, and a flow sensor 1916. The flexible bag 1900 may be a four-port bag as discussed above with reference to bag 1800 in FIG. 18 , and the bag 1900 and its components may share any one or more characteristics in common with bag 1800 and its individual components. FIGS. 19A-19D illustrate the bag 1900 in four different phases of cell processing using the bag 1900 in a system for delivering a payload to cells. The four phases are discussed below.

[0363] 19A shows bag 1900 while it is being filled with product (e.g., filled with cell suspension liquid). As shown, sample product may enter bag 1900 through product inlet 1902, and air and / or other gases may exit bag 1900 through air inlet 1904 as bag 1900 fills with liquid and air / gas is displaced. Flow through inlet 1902 and / or through air inlet 1904 may be controlled by one or more manually and / or automatically controlled valves or flow control mechanisms.

[0364] 19B shows bag 1900 while the product (e.g., cell suspension liquid) within bag 1900 is full and being cooled and / or otherwise prepared for passage through the constriction cartridge. As shown, pump 1914 can be activated, causing product to be drawn into and circulated through circulation loop 1908, traveling counterclockwise around pump 1914, past flow sensor 1916, upward out of the Y-junction / T-junction, toward and back into the main body of bag 1900. During this circulation process, flow of product downward out of the Y-junction / T-junction (e.g., toward and into a filter, constriction cartridge, and / or other downstream component) can be prevented by closure of one or more manually and / or automatically controlled valves or flow control mechanisms, which can be positioned downstream of the Y-junction / T-junction and upstream of the filter, constriction cartridge, and / or other downstream component.

[0365] 19C shows bag 1900 while product (e.g., cell suspension liquid) is flowing toward, into, and / or through the constriction cartridge following cooling and / or other preparation. As shown, pump 1914 may remain activated, and some product may continue to be drawn into and circulated through circulation loop 1908, progressing counterclockwise around pump 1914, past flow sensor 1916, upward out of the Y-junction / T-junction, and back toward and into the main body of bag 1900. However, some product may also flow downward from the Y-junction / T-junction toward, into, and / or through the constriction cartridge (or other downstream component). In some embodiments, product flow downward from the Y-junction / T-junction (e.g., toward and into a filter, constriction cartridge, and / or other downstream component) may be enabled by the opening of one or more manually and / or automatically controlled valves or flow control mechanisms. In some embodiments, the circulation rate for the circulation loop may be higher than the rate of flow through the filter, constriction cartridge, or other downstream component. Thus, a majority of the product may flow upward from the Y-junction / T-junction, while a small portion of the product may flow downward from the Y-junction / T-junction.

[0366] 19D shows bag 1900 at the end of a process for flowing product (e.g., cell suspension liquid) toward, into, and / or through the constriction cartridge. In some embodiments, flow sensor 1916 may detect when air or other gas enters circulation loop 1908, thereby indicating that the main body of bag 1900 is empty or nearly empty. In response to detecting air entering circulation loop 1908, the system within which bag 1900 is placed may automatically deactivate pump 1914, allowing any remaining sample in the return path to be expelled downward, out outlet 1906, toward and into downstream components. Shutting off pump 1914 in this manner may prevent bubbles from being recirculated into the main body of bag 1900 when the bag 1900 empties of liquid prematurely.

[0367] Although FIGS. 19A-19D show a four-port bag, circulation of a sample through a circulation loop may also be implemented using a five-port bag, such as bag 1700 shown in FIG. 17, in which the circulation loop therein is separate from the output flow path. In a five-port bag, one or more flow sensors may monitor the flow of fluid within the flow path of the circulation loop (e.g., loop 1708) and determine when fluid has drained from the bag to a sufficiently low level that fluid is no longer circulating through the loop (e.g., the fluid level has dropped below the circulation loop). In response to determining that fluid is no longer circulating through the loop, the pump causing fluid flow through the loop may be deactivated so that the remaining fluid may be drained through the output flow path. Alternatively, or in addition, one or more flow sensors may monitor the flow of fluid through the output flow path, and circulation of fluid through the circulation loop may be stopped only when the flow sensor monitoring the flow in the output flow path determines that fluid is no longer flowing through the output flow path, thereby indicating that the bag has emptied.

[0368] In some embodiments, using any one or more of the systems or methods disclosed herein may enable improved throughput for processing cells to deliver payloads to cells. In some embodiments, a system such as any one or more of the systems disclosed herein (e.g., System 100, System 1500, etc.) may be configured to process more than 1 billion cells / minute, 10 billion cells / minute, 100 billion cells / minute, 1 trillion cells / minute, or 10 trillion cells / minute. In some embodiments, a system such as any one or more of the systems disclosed herein (e.g., System 100, System 1500, etc.) may be configured to process less than 1 billion cells / minute, 10 billion cells / minute, 100 billion cells / minute, 1 trillion cells / minute, or 10 trillion cells / minute. In some embodiments, a system such as any one or more of the systems disclosed herein (e.g., System 100, System 1500, etc.) may be configured to process more than 1 billion cells / system step (e.g., per hour using the system without changing input bags and / or refilling preparation vessels), 10 billion cells / system step, 100 billion cells / system step, 1 trillion cells / system step, 10 trillion cells / system step, or 100 trillion cells / system step. In some embodiments, a system such as any one or more of the systems disclosed herein (e.g., System 100, System 1500, etc.) may be configured to process less than 1 billion cells / system step, 10 billion cells / system step, 100 billion cells / system step, 1 trillion cells / system step, 10 trillion cells / system step, or 100 trillion cells / system step. In some embodiments, the throughput rate / unit time and / or throughput capacity / system step may depend on cell size. For example, smaller cells such as red blood cells can be processed at a faster rate than larger cells such as peripheral blood mononuclear cells.

[0369] In some embodiments, any one or more of the sensors described herein (e.g., flow sensors, temperature sensors, pressure sensors, etc.) may be provided as an integrated part of a system, such as any one or more of the systems disclosed herein (e.g., system 100, system 1500, etc.). In some embodiments, alternatively or additionally, any one or more of the sensors described herein (e.g., flow sensors, temperature sensors, pressure sensors, etc.) may be provided as part of a removable, replaceable, modular, and / or disposable component, such as a disposable assembly, that is configured to be inserted into, electronically and / or physically coupled with, and / or otherwise interact with, any one or more of the systems disclosed herein (e.g., system 100, system 1500, etc.). computer

[0370] 20 illustrates an example of a computer, according to some embodiments. Computer 2000 can be a component of any of the systems or electronic devices described herein. For example, computer 2000 can be a computing device included in system 100, system 1000, system 1100, system 1500, system 1600, and / or any associated electronic device and / or any other electronic device disclosed herein. In some embodiments, computer 2000 can be configured to perform all or a portion of any of the methods described herein, such as all or a portion of method 1300 or 1400.

[0371] The computer 2000 can be a host computer connected to a network. The computer 2000 can be a client computer or a server. As shown in Figure 20, the computer 2000 can be any suitable type of microprocessor-based device, such as a personal computer, a workstation, a server, or a handheld computing device such as a phone or tablet. The computer can include, for example, one or more of a processor 2010, an input device 2020, an output device 2030, a storage device 2040, and a communication device 2060.

[0372] The input device(s) 2020 can be any suitable device that provides input, such as a touchscreen or monitor, a keyboard, a mouse, or a voice recognition device. The output device(s) 2030 can be any suitable device that provides output, such as a touchscreen, a monitor, a printer, a disk drive, or a speaker.

[0373] The storage device 2040 can be any suitable device providing storage, such as electrical, magnetic, or optical memory, including RAM, cache, hard drive, CD-ROM drive, tape drive, or removable storage disk. The communication device 2060 can include any suitable device capable of transmitting and receiving signals over a network, such as a network interface chip or card. The components of a computer can be connected in any suitable manner, such as via a physical bus or wirelessly. The storage device 2040 can be a non-transitory computer-readable storage medium comprising one or more programs that, when executed by one or more processors, such as processor 2010, cause the one or more processors to perform all or a portion of any of the methods or techniques described herein, such as all or a portion of method 1300 or 1400.

[0374] The software 2050, which may be stored in the memory device 2040 and executed by the processor 2010, may include, for example, programming that embodies the functionality of the present disclosure (e.g., as embodied in the systems, computers, servers, and / or devices as described above). In some embodiments, the software 2050 may be implemented and executed on a combination of servers, such as an application server and a database server.

[0375] The software 2050 can also be stored and / or carried within any computer-readable storage medium for use by or in connection with an instruction execution system, apparatus, or device, such as those described above, that can fetch and execute instructions associated with the software from the instruction execution system, apparatus, or device. In the context of the present disclosure, a computer-readable storage medium can be any medium, such as storage device 2040, that can contain or store programming for use by or in connection with a system, apparatus, or device.

[0376] The software 2050 can also be propagated in any carrier medium for use by or in connection with an instruction execution system, apparatus, or device, such as those described above, that can fetch and execute instructions associated with the software from the instruction execution system, apparatus, or device. In the context of this disclosure, a carrier medium can be any medium that can communicate, propagate, or carry programming for use by or in connection with an instruction execution system, apparatus, or device. Carrier-readable media can include, but are not limited to, electronic, magnetic, optical, electromagnetic, or infrared wired or wireless propagation media.

[0377] The computer 2000 may be connected to a network, which may be any suitable type of interconnected communications system. The network may implement any suitable communications protocol and may be secured by any suitable security protocol. The network may comprise any suitable arrangement of network links that may implement the transmission and reception of network signals, such as wireless network connections, T1 or T3 lines, cable networks, DSL, or telephone lines.

[0378] Computer 2000 can implement any operating system suitable for operation over a network. Software 2050 can be written in any suitable programming language, such as C, C++, Java, or Python. In various embodiments, application software embodying functionality of the present disclosure can be deployed in different configurations, such as, for example, in a client / server arrangement, or through a web browser as a web-based application or web service. [Embodiment]

[0379] Below is an enumerated list of certain embodiments. In some embodiments, features of any one or more of the following embodiments may be combined with any one or more of the other embodiments, even if the embodiment dependencies do not explicitly indicate that the embodiments may be combined. (Embodiment 1) 1. A system for delivering a payload to a cell, comprising: a holder configured to hold a cell suspension input container containing a cell suspension comprising cells; a receptacle configured to receive all or a portion of a disposable assembly, the disposable assembly comprising: a preparation vessel in fluid communication with the input container and configured to hold a cell suspension as it is prepared for passage through one or more cell-deforming constrictions; a constriction cartridge configured to be in fluid communication with the preparation vessel, the constriction cartridge configured to house a component comprising one or more cell-deforming constrictions, the cell-deforming constrictions configured to create a perturbation in a cell membrane to allow entry of a payload into the cell; a receiver comprising: one or more processors configured to receive input from a user and control one or more control modules configured to control one or more of pressure, temperature, agitation, and flow of the cell suspension, the one or more control modules a flow control module configured to flow the cell suspension from the input container, through the disposable assembly, to the cell suspension output container such that the payload is delivered into the cells. one or more processors comprising: A system comprising a platform supporting the system. (Embodiment 2) a microfluidic chip, wherein a component comprises a microfluidic channel, the microfluidic channel comprising one or more cell deformation constrictions; The constriction cartridge is a microfluidic chip cartridge configured to store a microfluidic chip; 2. The system of embodiment 1. (Embodiment 3) a filter comprising one or more cell deformation constrictions, wherein the component comprises a plurality of cell deformation constrictions; The constriction cartridge is a filter cartridge configured to house a filter; 3. A system according to any one of embodiments 1 and 2. (Embodiment 4) A system described in any one of embodiments 1-3, wherein the platform is a tabletop platform. (Embodiment 5) 5. The system of any one of embodiments 1-4, wherein the cell suspension comprises a payload. (Embodiment 6) 6. The system of any one of embodiments 1-5, wherein the system is configured to contact the payload with the cell suspension prior to flow of at least a portion of the cell suspension through the constriction cartridge. (Embodiment 7) 7. The system of any one of embodiments 1-6, wherein the system is configured to contact the payload with the cell suspension following flow of at least a portion of the cell suspension through the constriction cartridge. (Embodiment 8) 8. The system of any one of embodiments 1-7, wherein the one or more processors are configured to apply pressure to the cell suspension inside the preparation vessel. (Embodiment 9) Embodiment 10. The system of embodiment 8, wherein the one or more control modules comprise a pressure control module comprising a pressure source configured to deliver sterilizing gas to the preparation vessel. 10. The system according to any one of embodiments 8 and 9, wherein the pressure applied to the cell suspension inside the preparation vessel is sufficient to bring the preparation vessel into contact with the inner wall of the receiver. (Embodiment 11) 11. The system of any one of embodiments 8-10, wherein the pressure applied to the cell suspension inside the preparation vessel is sufficient to force the cell suspension out of the preparation vessel through the constriction cartridge. (Embodiment 12) 12. The system of any one of embodiments 1-11, wherein the one or more processors are configured to heat or cool the cell suspension inside the preparation vessel. (Embodiment 13) 13. The system of embodiment 12, wherein the one or more control modules comprise a temperature control module comprising one or more thermoelectric temperature control devices configured to heat or cool a portion of the receiver configured to contact the preparation vessel. (Embodiment 14) 14. The system of embodiment 13, wherein the portion of the receiver configured to contact the preparation vessel is a conductive outer jacket configured to conduct heat to and from the preparation vessel. (Embodiment 15) 15. The system of any one of embodiments 12-14, wherein the one or more control modules comprise a temperature control module comprising one or more thermoelectric temperature control devices configured to heat or cool the preparation vessel. (Embodiment 16) 16. The system of embodiment 15, wherein the one or more thermoelectric temperature control devices comprise a cooling plate disposed within a wall of the receiver and configured to contact an outer wall of the preparation vessel when the preparation vessel is inserted into the receiver. (Embodiment 17) 17. The system of any one of embodiments 1-16, wherein the one or more processors are configured to agitate the cell suspension inside the preparation vessel so that the cells are homogeneously dispersed in the cell suspension. (Embodiment 18) 18. The system of embodiment 17, wherein the one or more control modules comprise an agitation control module comprising a stirring plate configured to be driven by one or more motors, the stirring plate configured to cause agitation of all or a portion of the receiver. (Embodiment 19) A system described in any one of embodiments 1-18, wherein the flow control module is configured to cause one or more valves to control the flow of the cell suspension from an input container, through a disposable assembly, to an output container. (Embodiment 20) A system described in any one of embodiments 1-19, wherein the flow control module is configured to flow the cell suspension at a target fluid velocity. (Embodiment 21) A system described in any one of embodiments 1-20, further comprising an input device configured to receive instructions from a user, and wherein the one or more processors are configured to operate one or more of the control modules in response to the instructions. (Embodiment 22) 22. The system of embodiment 21, wherein the input device comprises a touchscreen display configured to transmit a signal to one or more of the control modules in response to detecting contact by a user. (Embodiment 23) 23. The system of any one of embodiments 21 and 22, wherein the input device comprises an agitation speed adjustment device configured to control the speed of a motor that causes agitation of the cell suspension inside the preparation vessel. (Embodiment 24) 24. The system of any one of embodiments 1-23, wherein the preparation vessel is configured to hold up to 10 liters of cell suspension. (Embodiment 25) 25. A system according to any one of embodiments 1-24, wherein the preparation vessel is configured to allow a pressure of up to 120 psi to be applied to the cell sus...

Claims

1. A system for delivering a payload to a cell, the system comprising: a disposable assembly attachable to the system inside a receptacle or housing, the disposable assembly comprising: a preparation vessel configured to hold a cell suspension as the cell suspension is prepared for passage through one or more cell-deforming constrictions; a constriction cartridge fluidly connected to the preparation vessel via an inlet, the constriction cartridge comprising a housing portion configured to adjustably or replaceably store and retain components comprising the one or more cell-deforming constrictions, the one or more cell-deforming constrictions having a diameter smaller than a diameter of the cells and configured to create a perturbation in a cell membrane of the cells to allow entry of a payload into the cells; a disposable assembly comprising: one or more control modules including a pressure control module that causes pressure to be applied to the cell suspension, and optionally one or more other control modules configured to control one or more of temperature, agitation, and flow of the cell suspension; wherein the constriction cartridge is configured such that the components are held between a base portion and a removable lid portion.

2. The system described in claim 1, further comprising one or more processors configured to receive input from a user and control the pressure control module, and optionally, the one or more other control modules configured to control one or more of the pressure, temperature, agitation, and flow of the cell suspension. (a) the component comprising the one or more cell-deforming constrictions is a microfluidic chip comprising a microfluidic channel, the microfluidic channel comprising the one or more cell-deforming constrictions, and the constriction cartridge is a microfluidic chip cartridge configured to store the microfluidic chip; or 3. The system of claim 1 or claim 2, wherein (b) the component comprising the one or more cell deforming constrictions is a filter comprising a plurality of cell deforming constrictions, and the constriction cartridge is a filter cartridge configured to house the filter.

4. A system described in any one of claims 1 to 3, wherein the system is configured to contact the payload with the cell suspension before or after flow of at least a portion of the cell suspension through the constriction cartridge.

5. The one or more processors: a) causing pressure to be applied to the cell suspension inside the preparation vessel, the pressure applied to the cell suspension inside the preparation vessel being sufficient to cause the cell suspension to be forced through the constriction cartridge and out of the preparation vessel; and / or b) receiving signals from one or more sensors and automatically controlling one or more of the pressure, temperature, and agitation of the cell suspension according to the received signals; The system according to any one of claims 2 to 4, configured to:

6. The one or more sensors (a) a temperature sensor configured to monitor the temperature of the cell suspension; (b) a bubble sensor configured to monitor the flow of the cell suspension; (c) a pressure sensor configured to monitor the pressure applied to the cell suspension; and / or (d) a pressure sensor configured to monitor the pressure inside the preparation vessel; The system of claim 5 , comprising:

7. The one or more other control modules: a) a temperature control module comprising one or more thermoelectric temperature control devices configured to heat or cool a portion of the receiver configured to contact the preparation vessel; and / or b) a flow control module configured to cause the cell suspension to flow from the input container through the disposable assembly to the cell suspension output container such that the payload is delivered into the cells, the flow control module configured to cause one or more valves to control the flow of the cell suspension from the input container through the disposable assembly to the output container. The system of claim 1 , comprising:

8. A system described in any of claims 2 to 7, further comprising an input device configured to receive instructions from the user, wherein the one or more processors are configured to operate one or more of the control modules in response to the instructions.

9. The preparation vessel comprises: (a) a first inlet configured to be fluidly connected to a cell suspension input container to receive a flow of the cell suspension, and an outlet configured to be fluidly connected to the constriction cartridge; and / or (b) a second inlet configured to be fluidly connected to a pressure source to receive a flow of sterilizing gas into the preparation vessel to cause pressure to be applied to the cell suspension; The system according to any one of claims 1 to 8, comprising:

10. A system described in any of claims 1 to 9, wherein the housing portion is configured to direct the flow of the cell suspension through the component comprising the one or more cell-deforming constrictions.

11. A system as described in any one of claims 1 to 10, wherein the constriction cartridge has an outlet configured to be fluidly connected to an output container.

12. A system described in any one of claims 1 to 11, further comprising a filter configured to receive fluid flow downstream from the preparation vessel and upstream of the constriction cartridge, the filter configured to remove multicellular aggregates from the cell suspension before it reaches the constriction cartridge.

13. A system described in any one of claims 1 to 12, wherein the preparation vessel is provided with a circulation loop configured to draw liquid from a main body of the preparation vessel and circulate the liquid back into the main body of the preparation vessel.

14. The system described in claim 13, wherein a portion of the circulation loop is integrated with a flow path leading from the preparation vessel to the constriction cartridge.

15. A method for delivering a payload to a cell, said method comprising: providing cells in a cell suspension; transferring the cell suspension into a preparation vessel; preparing the cell suspension, including by causing pressure to be applied to the cell suspension while the cell suspension is in the preparation vessel; passing the prepared cell suspension from the preparation vessel through a constriction cartridge of the system, the constriction cartridge comprising a housing portion configured to adjustably or replaceably store and retain components comprising a cell deformation constriction, the cell deformation constriction having a diameter smaller than a diameter of the cells, causing a perturbation in a cell membrane of the cells to allow entry of a payload into the cells; wherein the constriction cartridge is configured such that the components are held between a base portion and a removable lid portion. (a) the component comprising the one or more cell-deforming constrictions is a microfluidic chip comprising a microfluidic channel, the microfluidic channel comprising the one or more cell-deforming constrictions, and the constriction cartridge is a microfluidic chip cartridge configured to store the microfluidic chip; or 16. The method of claim 15, wherein (b) the component comprising the one or more cell deforming constrictions is a filter comprising a plurality of cell deforming constrictions, and the constriction cartridge is a filter cartridge configured to house the filter.

17. The method of claim 15 or 16, further comprising attaching a disposable assembly comprising the preparation container and the constriction cartridge to the system.

18. The method of claim 17, wherein attaching the disposable assembly includes attaching one or more sensors included in the disposable assembly so that the sensors are configured to transmit signals to the system.

19. A method described in any one of claims 15 to 18, comprising performing an integrity check on the disposable assembly before transferring the cell suspension into the disposable assembly.

20. The method of claim 19, wherein performing an integrity check on the disposable assembly includes pressurizing a gas inside the disposable assembly and monitoring the pressure of the pressurized gas over a predetermined period of time.

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