Simultaneous maximization of cell permeabilization and viability in single cell electroporation using an electrolyte-filled capillary

摘要

A549 cells were briefly exposed to Thioglo −1 which converts thiols to fluorescent adducts. The fluorescent cells were exposed to short (50 − 300 ms) electric field pulses (500 V across a 15 cm capillary) created at the tip of an electrolyte - filled capillary. Fluorescence microscopy revealed varying degrees of cell permeabilization depending on conditions. Longer pulses and shorter cell - capillary tip distance led to a greater decrease in a cell's fluorescence. Live/dead (calcein AM and propidium iodide) testing revealed that a certain fraction of cells died. Longer pulses and shorter cell – capillary tip distances were more deadly. An optimum condition exists at a cell – capillary tip distance of 3.5 μm − 4.5 μm and a pulse duration of 120 ms - 150 ms. At these conditions > 90 % of the cells are permeabilized and 80 − 90% survive.