US 11,725,175 B2
Electroporation
Viktor Shkolnikov, Palo Alto, CA (US); Diane R. Hammerstad, Corvallis, OR (US); and Michael W. Cumbie, Corvallis, OR (US)
Assigned to Hewlett-Packard Development Company, L.P., Spring, TX (US)
Appl. No. 16/605,193
Filed by HEWLETT-PACKARD DEVELOPMENT COMPANY, L.P., Spring, TX (US)
PCT Filed Sep. 23, 2017, PCT No. PCT/US2017/053121
§ 371(c)(1), (2) Date Oct. 14, 2019,
PCT Pub. No. WO2019/059936, PCT Pub. Date Mar. 28, 2019.
Prior Publication US 2020/0224145 A1, Jul. 16, 2020
Int. Cl. C12M 1/42 (2006.01); C12M 1/26 (2006.01); C12M 1/36 (2006.01); C12N 13/00 (2006.01); C12N 15/87 (2006.01); C12M 3/00 (2006.01); A61N 1/10 (2006.01); C12M 1/00 (2006.01)
CPC C12M 35/02 (2013.01) [C12M 1/42 (2013.01); C12M 3/006 (2013.01); C12M 33/04 (2013.01); C12M 41/48 (2013.01); C12N 13/00 (2013.01); C12N 15/87 (2013.01); A61N 1/10 (2013.01); C12M 45/07 (2013.01)] 18 Claims
OG exemplary drawing
 
1. An electroporation system comprising:
a well plate comprising:
wells, each of the wells comprising:
an interior formed by first and second sidewalls and a floor;
a first electrode adjacent the interior and which extends along an interior side surface of the first sidewall; and
a second electrode adjacent the interior and which extends along an interior side surface of the second sidewall, and the second electrode being spaced from the first electrode by the floor, and the first electrode and the second electrode to apply an electrostatic field across the well;
a well support to carry the well plate, the well support including at least one datum surface to contact the well plate to position each of the wells of the well plate;
a dispenser including a plurality of reservoirs containing different fluids and a plurality of fluid jetting devices to selectively dispense the different fluids, including a cell having a diameter, into each of the wells; and
a dispenser-well positioning system including a controller and actuator, wherein the controller is communicatively coupled to the actuator and programmed to align each well and the dispenser based upon a position of the at least one datum surface and a position of the dispenser such that the dispenser dispenses the cell into each well at a location spaced from the first electrode and the second electrode by a distance of at least 5 times the diameter of the cell.
 
13. An electroporation system comprising:
a well plate comprising:
wells, each well comprising:
an interior formed by first and second sidewalls and a floor;
a first electrode adjacent the interior and which extends along an interior side surface of the first sidewall; and
a second electrode adjacent the interior and which extends along an interior side surface of the second sidewall, and the second electrode being spaced from the first electrode by the floor, and the first electrode and the second electrode to apply an electrostatic field across the well;
a well support to carry the well plate, the well support including at least one datum surface to contact the well plate to position each of the wells of the well plate;
a plurality of reservoirs containing different fluids including a plurality of different types of cells;
a cell dispenser coupled to the plurality of reservoirs and including a plurality of fluid jetting devices;
at least one actuator operably coupled to the well plate and the cell dispenser; and
a controller programmed to output control signals to:
cause the at least one actuator to align each well and the cell dispenser based upon a position of the at least one datum surface and a position of the cell dispenser and to cause the cell dispenser to selectively dispense a different type of cell among the plurality of different types of cells from the plurality of reservoirs into each of the wells; and
form an electrostatic field within each of the wells;
wherein the actuator and the controller form part of a dispenser-well positioning system programmed to align each well of the wells and the cell dispenser such that the cell dispenser dispenses the cell into each well at a location spaced from the first electrode and the second electrode by a distance of at least 10 times a diameter of the cell.
 
15. An electroporation system comprising:
a well plate comprising:
wells, each well comprising:
an interior formed by first and second sidewalls and a floor;
a first electrode adjacent the interior and which extends along an interior side surface of the first sidewall; and
a second electrode adjacent the interior and which extends along an interior side surface of the second sidewall, and the second electrode being spaced from the first electrode by the floor, and the first electrode and the second electrode to apply an electrostatic field across the well;
a well support to carry the well plate, the well support including at least one datum surface to contact the well plate to position each of the wells of the well plate;
a plurality of reservoirs containing different fluids including a plurality of different types of cells;
a cell dispenser coupled to the plurality of reservoirs and including a plurality of fluid jetting devices;
at least one actuator operably coupled to the well plate and the cell dispenser; and
a controller programmed to output control signals to:
cause the at least one actuator to align each well and the cell dispenser based upon a position of the at least one datum surface and a position of the cell dispenser and to cause the cell dispenser to selectively dispense a different type of cell among the plurality of different types of cells from the plurality of reservoirs into each of the wells; and
form an electrostatic field within each of the wells;
wherein the controller is programmed to:
cause selective dispense of M different types of cells, N different types of transfection agents, and R different electroporation solutions to the wells, and form P different electrostatic fields, wherein the different fluids include the M different types of cells, the N different types of transfection agents, and the R different electroporation solution.