Method and apparatus for electroporation of acoustically-aligned cells

Patent metadata

Field	Value
Publication number	US-11591561-B2
Application number	US-201816168464-A
Country	US
Kind code	B2
Filing date	Oct 23, 2018
Priority date	Oct 23, 2017
Publication date	Feb 28, 2023
Grant date	Feb 28, 2023

Abstract

Transfer of genetic and other materials to cells is conducted in a hands-free, automated and continuous process that includes flowing the cells between electroporation electrodes to facilitate delivery of a payload into the cells, while acoustophoretically focusing the cells. Also described is a control method for the acoustophoretic focusing of cells that includes detecting locations of cells flowing through a channel, such as with an image analytics system, and modulating a drive signal to an acoustic transducer to change the locations of the cells flowing in the channel. Finally, an electroporation driver module is described that uses a digital to analog converter for generating an electroporation waveform and an amplifier for amplifying the electroporation waveform for application to electroporation electrodes.

First claim

What is claimed is:

1. A method for introducing a payload into cells, the method comprising:
flowing the cells between electroporation electrodes to facilitate delivery of the payload into the cells by flowing the cells through a microchannel system that has a main channel ending in a trifurcating outlet in which the cells are directed to a cell output channel that couples with a center of the main channel, and
sensing the presence of cells by capturing images of the cells in the microchannel system and analyzing the images to determine locations of the cells, while
acoustophoretically focusing the cells in a center of the main channel by modulating a drive signal in response to the analysis of the images.

2. The method of claim 1 , wherein acoustophoretically focusing the cells comprises driving an acoustic transducer that is attached to a substrate holding the electroporation electrodes.

3. The method of claim 1 , further comprising flowing spent cargo/solution downstream of the electroporation electrodes into two or more channels.

4. The method of claim 1 , further comprising driving the electroporation electrodes with an arbitrary waveform generator.

5. The method of claim 1 , wherein the presence of the cells is detected upstream of the electroporation electrodes.

6. The method of claim 1 , further comprising energizing the electroporation electrodes in response to sensing the presence of cells.

7. The method of claim 1 , further comprising measuring a current flow between the electroporation electrodes.

8. The method of claim 1 , further comprising flowing the cells in the microchannel system that includes a substrate having the main channel ending in the trifurcating outlet.

9. The method of claim 8 , further comprising acoustophoretically focusing the cells with an acoustic transducer that is attached to the substrate.

10. The method of claim 8 , wherein the microchannel system has two or more channels of the outlet for receiving spent cargo/solution downstream of the electroporation electrodes.

11. The method of claim 1 , further comprising analyzing the images to determine the locations of the cells in an inter-electrode region between the electroporation electrodes.

12. The method of claim 1 , further comprising the microchannel system establishing a sheath flow of a cell incubation fluid on either side of an electroporation fluid between the electroporation electrodes.

Assignees

• Charles Stark Draper Laboratory Inc

Inventors

• Tandon Vishal
• Lissandrello Charles A
• Dubay Ryan A
• Christianson Rebecca
• Balestrini Jenna Leigh
• Hsi Peter
• Fiering Jason

Classifications

• A61N7/00

  Ultrasound therapy (lithotripsy A61B17/22, A61B17/225; massage using supersonic vibration A61H23/00 {; using ultrasound for introducing media into the body A61M37/0092}) · CPC title

• C12M23/16

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

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12M35/02 Primary

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

• A61K9/0009

  involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis · CPC title