Measurement of transient permeability of Sp2/0 myeloma cells: flow cytometric study

Abstract

Electroporation is an electric field induced phenomenon occurring when the permeability of the cell membrane is increased due to the excess of critical transmembrane potential. Fluorescent dye assays are frequently used for evaluation of the permeabilization rate, however, the protocols vary, which negatively affects the repeatability of the results. In this work we have designed experiments to investigate the protocols and threshold concentrations of the Propidium Iodide (PI) and YO -PRO -1 (YP) fluorescent dyes for evaluation of mammalian cell permeabilization induced by electroporation. The Sp2/0 mouse myeloma cells were used and the bursts of 100 μs x 8 electrical pulses of 0.8 –2 kV/cm were applied. It has been shown that the dye concentration has an influence on the detectable permeabilization, and the concentrations below 30 μM for PI and 1 μM for YP should be avoided for measurement of electro - permeabilization efficacy due to unreliable fluorescence signals. Further, based on the experimental data, the permeabilization curve for the S p2/0 myeloma cells in the 0.8 –2 kV/cm range has been presented.