High throughput, feedback-controlled electroporation microdevice for efficient molecular delivery into single cells

Patent metadata

Field	Value
Publication number	US-11987777-B2
Application number	US-202117143659-A
Country	US
Kind code	B2
Filing date	Jan 7, 2021
Priority date	Sep 4, 2015
Publication date	May 21, 2024
Grant date	May 21, 2024

Abstract

Systems and methods for cell electroporation and molecular delivery using an intelligent, feedback controlled, microscale electroporation system for transfecting single cells.

First claim

What is claimed is:

1. A method for electroporating a plurality of biological cells, the method comprising:
hydrodynamically focusing a continuous flow of a plurality of biological cells into a single-file flow, wherein the single-file flow passes each of the plurality of biological cells through a detection area;
generating a cell detection signal;
continuously monitoring an impedance value of the detection area;
generating, in response to an increase in the impedance value of the detection area over a baseline threshold, a permeabilization signal, the baseline threshold being indicative of a presence of one of the plurality of biological cells within the detection area;
stopping the permeabilization signal in response to determining that the impedance value of the detection area is equal to or greater than a permeabilization threshold value; and
generating a delivery signal, wherein the delivery signal causes delivery of a molecule into one of the plurality of biological cells focused within the detection area.

2. The method according to claim 1 , further comprising adjusting at least one parameter of the permeabilization signal in response to determining that the impedance value of the detection area is less than the permeabilization threshold value.

3. The method according to claim 1 , further comprising:
determining whether the impedance value of the detection area is equal to the permeabilization threshold value; and
adjusting at least one parameter of the delivery signal in response to determining the impedance value of the detection area is not equal to the permeabilization threshold value.

4. The method according to claim 3 , further comprising:
stopping the delivery signal in response to determining the impedance value of the detection area is one of: less than or equal to a viability threshold, or equal to the baseline threshold.

5. The method according to claim 4 , further comprising stopping the delivery signal in response to at least one of the following:
determining that the impedance value is less than the threshold impedance indicating that that the single one of the plurality of biological cells has exited the detection area; or
determining that the impedance value is equal to a viability threshold for over-exposure.

6. The method according to claim 1 , wherein the baseline threshold is determined by continuously monitoring the impedance value of a second detection area through which only a buffer solution is flowing.

7. The method according to claim 2 , wherein the at least one parameter of the permeabilization signal is selected from the group of: electric field amplitude, pulse duration, pulse train frequency, duty cycle, and number of cycles.

8. The method according to claim 3 , wherein the at least one parameter of the delivery signal is selected from the group of: electric field amplitude, pulse duration, pulse train frequency, duty cycle, and number of cycles.

9. The method according to claim 1 , wherein the permeabilization threshold value is determined experimentally or using a mathematical model.

10. The method according to claim 1 , wherein the permeabilization threshold value corresponds to an optimal cell permeabilization that does not cause cell death.

Assignees

• Univ Rutgers

Inventors

• Zahn Jeffrey
• Zheng Mingde
• Shreiber David I
• Lin Hao
• Shan Jerry W

Classifications

• C12M1/42 Primary

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

• B01L3/502715 Primary

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• C12M23/16

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• C12M35/02 Primary

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