Method and Apparatus for Electroporation of Acoustically-Aligned Cells

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.

What is claimed is: 
     
         1 . 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.   
     
     
         2 . The device of  claim 1 , 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. 
     
     
         3 . The device of  claim 1 , wherein the acoustic transducer is attached to the substrate. 
     
     
         4 . The device of  claim 1 , wherein the microchannel system has two or more channels for receiving spent cargo/solution downstream of the electroporation electrodes. 
     
     
         5 . The device of  claim 1 , further comprising an arbitrary waveform generator for driving the electroporation electrodes. 
     
     
         6 . The device of  claim 1 , further comprising sense electrodes and a sensing module for detecting the presence of cells in the microchannel system. 
     
     
         7 . The device of  claim 6 , wherein sense electrodes are located upstream of the electroporation electrodes. 
     
     
         8 . The device of  claim 6 , further comprising a controller that energizes the electroporation electrodes in response to sensing the presence of cells. 
     
     
         9 . 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.   
     
     
         10 . A method as claimed in  claim 9 , 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.   
     
     
         11 . A method as claimed in  claim 10 , wherein the images are captured of an inter-electrode region between electroporation electrodes. 
     
     
         12 . A method as claimed in  claim 10 , wherein modulating the drive signal comprising modulating a frequency and/or amplitude to center the cells on the channel. 
     
     
         13 . 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.   
     
     
         14 . A device as claimed in  claim 13 , further comprising an image analytics system for analyzing the images to determine the locations of the cells. 
     
     
         15 . A device as claimed in  claim 13 , wherein the image capture device captures images of an inter-electrode region between electroporation electrodes. 
     
     
         16 . A device as claimed in  claim 13 , wherein the controller modulates a frequency and/or amplitude of the drive signal to center the cells on the channel. 
     
     
         17 . 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.   
     
     
         18 . 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.   
     
     
         19 . A system for movement of particles between fluid streams, comprising:
 a microchannel system that receives particles in a first fluid flowing through the microchannel device;   an acoustic transducer that generates acoustic radiation to move the particles to a second fluid flowing through the microchannel device.   
     
     
         20 . The system of  claim 19 , that controls residence times of particles in the streams. 
     
     
         21 . The system of  claim 19 , wherein the first fluid and the second fluid are merged in a laminar flow regime. 
     
     
         22 . The system of  claim 19 , wherein the first fluid is a cell incubation fluid and the second fluid is an electroporation fluid. 
     
     
         23 . The system of  claim 19 , further comprising an electroporation system in or downstream of the acoustic transducer. 
     
     
         24 . The system of  claim 19 , wherein the microchannel system establishes a sheath flow of the first fluid on either side of the second fluid.