Reversible electropermeabilization of mammalian cells by high-intensity, ultra-short pulses of submicrosecond duration.
Mouse myeloma cells were electropermeabilized by single square-wave electric pulses with amplitudes of up to approximately 150 kV/cm and durations of 10-100 nsec. The effects of the field intensity, pulse duration and medium conductivity on cell viability and field-induced uptake of molecules were analyzed by quantitative flow cytometry using the membrane-impermeable fluorescent dye propidium iodide as indicator molecule. Despite the extremely large field strengths, the majority of cells survived the exposure to ultra-short field pulses. The electrically induced dye uptake increased markedly with decreasing conductivity of the suspending medium. We assigned this phenomenon to the transient electrodeformation (stretching) force that assumes its maximum value if cells are suspended in low-conductivity media, i.e., if the external conductivity sigmae is smaller than that of the cytosol sigmai. The stretching force vanishes when sigmae is equal to or larger than sigmai. Due to their capability of delivering extremely large electric fields, the pulse power systems used here appear to be a promising tool for the electropermeabilization of very small cells and vesicles (including intracellular organelles, liposomes, etc.).
Müller
K J
KJ
Lehrstuhl für Biotechnologie, Biozentrum der Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Sukhorukov
V L
VL
Zimmermann
U
U
D000818:Animals; D002463:Cell Membrane Permeability / Q000502:physiology*; D004553:Electric Conductivity; D004558:Electric Stimulation / Q000379:methods; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D005470:Fluorometry / Q000379:methods; D051379:Mice; D009101:Multiple Myeloma; D011419:Propidium / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011419:Propidium