Simultaneous maximization of cell permeabilization and viability in single-cell electroporation using an electrolyte-filled capillary.
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 the conditions. Longer pulses and a shorter cell-capillary tip distance led to a greater decrease in the 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 3.5-4.5 microm and a pulse duration of 120-150 ms. At these conditions, >90% of the cells are permeabilized and 80-90% survive.
Agarwal
Aparna
A
Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.
Zudans
Imants
I
Orwar
Owe
O
Weber
Stephen G
SG
D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000502:physiology / Q000528:radiation effects*; D002470:Cell Survival / Q000502:physiology / Q000528:radiation effects; D004573:Electrolytes / Q000737:chemistry*; D004574:Electromagnetic Fields; D018274:Electroporation / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008856:Microscopy, Fluorescence; D009281:Naphthalenes / Q000737:chemistry; D011419:Propidium / Q000737:chemistry; D011758:Pyrroles / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry
D004573:Electrolytes; D005452:Fluoresceins; D005456:Fluorescent Dyes; D009281:Naphthalenes; D011758:Pyrroles; D013438:Sulfhydryl Compounds; C498335:ThioGlo-1; C085925:calcein AM; D011419:Propidium