Single-cell electroporation arrays with real-time monitoring and feedback control.
Rapid well-controlled intracellular delivery of drug compounds, RNA, or DNA into a cell--without permanent damage to the cell--is a pervasive challenge in basic cell biology research, drug discovery, and gene delivery. To address this challenge, we have developed a bench-top system comprised of a control interface, that mates to disposable 96-well-formatted microfluidic devices, enabling the individual manipulation, electroporation and real-time monitoring of each cell in suspension. This is the first demonstrated real-time feedback-controlled electroporation of an array of single-cells. Our computer program automatically detects electroporation events and subsequently releases the electric field, precluding continued field-induced damage of the cell, to allow for membrane resealing. Using this novel set-up, we demonstrate the reliable electroporation of an array (n = 15) of individual cells in suspension, using low applied electric fields (<1 V) and the rapid and localized intracellular delivery of otherwise impermeable compounds (Calcein and Orange Green Dextran). Such multiplexed electrical and optical measurements as a function of time are not attainable with typical electroporation setups. This system, which mounts on an inverted microscope, obviates many issues typically associated with prototypical microfluidic chip setups and, more importantly, offers well-controlled and reproducible parallel pressure and electrical application to individual cells for repeatability.
Khine
Michelle
M
School of Engineering, University of California, Merced, CA, USA. mkhine@ucmerced.edu
Ionescu-Zanetti
Cristian
C
Blatz
Andrew
A
Wang
Lee-Ping
LP
Lee
Luke P
LP
D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D016503:Drug Delivery Systems; D004563:Electrochemistry / Q000379:methods; D018274:Electroporation; D025461:Feedback, Physiological; D005452:Fluoresceins / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D044085:Microfluidics / Q000295:instrumentation*; D008904:Miniaturization; D012313:RNA / Q000737:chemistry; D012984:Software
D005452:Fluoresceins; D005456:Fluorescent Dyes; D012313:RNA; D004247:DNA; C007740:fluorexon