Characterization of Cell Membrane Permeability In Vitro Part I: Transport Behavior Induced by Single-Pulse Electric Fields.

Abstract

Most experimental studies of electroporation focus on permeabilization of the outer cell membrane. Some experiments address delivery of ions and molecules into cells that should survive; others focus on efficient killing of the cells with minimal temperature rise. A basic method for quantifying electroporation effectiveness is measuring the membrane’s diffusive permeability. More specifically, comparisons of membrane permeability between electroporation protocols often rely on relative fluorescence measurements, which are not able to be directly connected to theoretical calculations and complicate comparisons between studies. Here we present part I of a 2-part study: a research method for quantitatively determining the membrane diffusive permeability for individual cells using fluorescence microscopy. We determine diffusive permeabilities of cell membranes to propidium for electric field pulses with durations of 1 to 1000 μs and strengths of 170 to 400 kV/m and show that diffusive permeabilities can reach 1.3±0.4×10−8 m/s. This leads to a correlation between increased membrane permeability and eventual propidium uptake. We also identify a subpopulation of cells that exhibit a delayed and significant propidium uptake for relatively small single pulses. Our results provide evidence that cells, especially those that uptake propidium more slowly, can achieve large permeabilities with a single electrical pulse that may be quantitatively measured using standard fluorescence microscopy equipment and techniques.