Method and Apparatus for Electroporation of Acoustically-Aligned Cells

Patent metadata

Field	Value
Publication number	US-2019119624-A1
Application number	US-201816168464-A
Country	US
Kind code	A1
Filing date	Oct 23, 2018
Priority date	Oct 23, 2017
Publication date	Apr 25, 2019
Grant date	—

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, while acoustophoretically focusing the cells.

2 . The method of claim 1 , wherein flowing the cells comprises 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.

3 . 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.

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

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

6 . The method of claim 1 , further comprising sensing the presence of cells.

7 . The method of claim 6 , wherein the presence of the cells is detected upstream of the electroporation electrodes.

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

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

10 . A device for introducing a payload into cells, the device comprising:
a microchannel system for flowing the cells between electroporation electrodes to facilitate delivery of the payload into the cells; and an acoustic transducer for focusing the cells between the electroporation electrodes.

11 . The device of claim 10 , wherein the microchannel system includes a substrate having 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.

12 . The device of claim 10 , wherein the acoustic transducer is attached to the substrate.

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

14 . The device of claim 10 , further comprising an arbitrary waveform generator for driving the electroporation electrodes.

15 . The device of claim 10 , further comprising sense electrodes and a sensing module for detecting the presence of cells in the microchannel system.

16 . The device of claim 15 , wherein sense electrodes are located upstream of the electroporation electrodes.

17 . The device of claim 15 , further comprising a controller that energizes the electroporation electrodes in response to sensing the presence of cells.

18 . A control method for acoustophoretic focusing of cells, comprising:
detecting locations of cells flowing through a channel; and modulating a drive signal to an acoustic transducer to change the locations of the cells flowing in the channel.

19 . A method as claimed in claim 18 , wherein detecting locations of the cells comprises:
capturing images of the cells in the channel; and analyzing the images to determine the locations of the cells.

20 . A method as claimed in claim 19 , wherein the images are captured of an inter-electrode region between electroporation electrodes.

21 . A method as claimed in claim 19 , wherein modulating the drive signal comprising modulating a frequency and/or amplitude to center the cells on the channel.

22 . A device for processing cells, comprising:
an image capture device for capturing images of cells flowing through a channel; an acoustic transducer for acoustophoretic focusing the cells in the channel; and a controller for modulating a drive signal to the acoustic transducer to change the locations of the cells flowing in the channel based on the captured images.

23 . A device as claimed in claim 22 , further comprising an image analytics system for analyzing the images to determine the locations of the cells.

24 . A device as claimed in claim 22 , wherein the image capture device captures images of an inter-electrode region between electroporation electrodes.

25 . A device as claimed in claim 22 , wherein the controller modulates a frequency and/or amplitude of the drive signal to center the cells on the channel.

26 . An electroporation driver module, comprising:
A digital to analog converter for generating an electroporation waveform; and an amplifier for amplifying the electroporation waveform for application to electroporation electrodes.

27 . An electroporation driver module, comprising:
a first analog to digital converter for detecting a voltage applied to electroporation electrodes; and a second analog to digital converter for detecting a current flowing 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

• A61K9/0009

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

• C12M35/04

  Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli · 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

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title