Electroporation

Patent metadata

Field	Value
Publication number	US-11725175-B2
Application number	US-201716605193-A
Country	US
Kind code	B2
Filing date	Sep 23, 2017
Priority date	Sep 23, 2017
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

An electroporation system may include a well plate, a dispenser and a dispenser-well positioning system. The well plate may include wells, each of the wells including an interior, a first electrode adjacent the interior and a second electrode adjacent the interior and spaced from the first electrode. The first electrode and the second electrode are to apply an electrostatic field across the well. The dispenser is to dispense a cell having a diameter into each of the wells. The dispenser-well positioning system is to align each well and 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.

First claim

What is claimed is:

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.

2. The electroporation system of claim 1 , wherein each of the first and second electrodes of the wells are directly exposed to contents of the wells, wherein the different fluids include:
fluids containing different types of cells, different types of electroporation solutions, and different types of transfection agents.

3. The electroporation system of claim 1 , wherein each of the plurality of fluid jetting devices comprise a thermal resistive fluid actuator to eject the cell through a nozzle, and wherein the actuator of the dispenser-well positioning system is to move at least one of the dispenser and the well plate to position the nozzle of the dispenser at the position to dispense a cell into a respective well of the well plate at the location spaced from the first electrode and the second electrode of the respective well by the distance of at least 5 times the diameter of the cell.

4. The electroporation system of claim 1 , wherein the dispenser-well positioning system comprises:
the at least one datum surface to contact the well plate to position each of the wells of the well plate at a predetermined location;
the actuator to move the well support and the well plate relative to the dispenser; and
the controller programmed to control the actuator based upon a position of the at least one datum surface and a position of the dispenser to control positioning of the well plate relative to the dispenser.

5. The electroporation system of claim 4 , further comprising a second actuator to move the dispenser relative to the well support and the well plate, wherein the controller is programmed to control the second actuator based upon a position of the at least one datum surface.

6. The electroporation system of claim 1 , wherein the dispenser-well positioning system comprises:
the at least one datum surface to contact the well plate to position each of the wells of the well plate at a predetermined location, the well plate includes at least one member to contact against the at least one datum surface and to position the well plate;
the actuator to move the dispenser relative to the well support and the well plate; and
the controller is programmed to control the actuator based upon a position of the at least one datum surface and the position of the dispenser to control positioning of the dispenser relative to the well plate.

7. The electroporation system of claim 1 , wherein the dispenser-well positioning system comprises:
a sensor to sense the positioning of the well plate, wherein the sensor includes a photo emitter-detector arrangement or electrical contacts or switches;
the actuator operably coupled to at least one of the well plate and the dispenser to move said at least one of the well plate and the dispenser relative to one another; and
the controller is programmed to control the actuator based upon signals from the sensor indicating a sense positioning of the well plate.

8. The electroporation system of claim 1 , wherein the dispenser includes at least one of an impedance sensor and an optical sensor to count a number of cells dispensed into each of the wells.

9. The electroporation system of claim 1 , wherein the controller is programmed to:
differently charge the first electrode and the second electrode of each of the wells to form different P electrostatic fields within each of the wells based on cell types of cells within each of the wells;
cause the selective dispense of M different types of cells, selective dispense of 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 solutions; and
form a library of electrostatic field values for the different types of cells and different types of transfection agents based on a comparison of an agent transfection result for each of the wells.

10. The electroporation system of claim 1 , wherein the first electrode and the second electrode of each well respectively extend along the first and second sidewalls and out of each well and along an exterior surface of a member of the well plate to provide electrical connections, and the first and second electrodes are electrically isolated from one another and separated by the floor.

11. The electroporation system of claim 1 , wherein the at least one datum surface is to contact the well plate to position each of the wells of the well plate at a predetermined location, the at least one datum surface including:
at least one protuberance projecting along a top side of the well support; and
at least one detent that extends into the top side of the well support; and
the controller is programmed to control the actuator to move the well support relative to the dispenser based upon the position of the at least one datum surface and the position of the dispenser to control positioning of the well plate relative to the dispenser.

12. The electroporation system of claim 1 , wherein the at least one datum surface is selected from:
a protuberance projecting along a top side of the well support;
a detent that extends into the top side of the well support; and
a combination thereof, and
wherein the well plate includes at least one detent that extends into an underside of the well plate and a protuberance that extends from the underside of the well plate to abut the at least one datum surface and facilitate alignment of the dispenser with the wells of the well plate.

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 in

Assignees

• Hewlett Packard Development Co

Inventors

• Shkolnikov Viktor
• Hammerstad Diane R
• Cumbie Michael W

Classifications

• C12M35/02 Primary

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12M1/42

  Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12M3/006

  Cell injection or fusion devices · CPC title

• C12M33/04

  by injection or suction, e.g. using pipettes, syringes, needles (pipettes in general B01L3/02) · CPC title

• C12M41/48

  Automatic or computerized control (automatic analysis G01N35/00) · CPC title