96-Well single-cell electroporation arrays: Real-time monitoring and feedback control of membrane resealing kinetics

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, therapeutic gene delivery, and molecular medicine. To address this challenge, we have developed a bench-top system comprised of a control interface, with mating disposable 96-well-based microfluidic systems, that enables the manipulation, monitoring, and characterization of an array of single-cells. Using this set-up, we individually electroporate an array (n=15) of Hela cells in suspension with low applied voltages (0.4V-0.8V) and insert otherwise impermeable Calcein (Invitrogen, MW=622) and Orange Green Dextran 514 (Invitrogen, MW = 70,000) by electrophoresis (-200mV) within seconds.