We present experimental results regarding the effects of electric pulses on giant unilamellar vesicles (GUVs). We have used phase contrast and coherent anti-Stokes Raman scattering (CARS) microscopy as relevant optical approaches to gain insight into membrane changes under electropermeabilization. No addition of exogenous molecules (lipid analogue, fluorescent dye) was needed. Therefore, experiments were performed on pure lipid systems avoiding possible artefacts linked to their use. Structural membrane changes were assessed by loss of contrast inside the GUVs due to sucrose and glucose mixing. Our observations, performed at the single vesicle level, indicate these changes are under the control of the number of pulses and field intensity. Larger number of pulses enhances membrane alterations. A threshold value of the field intensity must be applied to allow exchange of molecules between GUVs and the external medium. This threshold depends on the size of the vesicles, the larger GUVs being affected at lower electric field strengths than the smaller ones. Our experimental data are well described by a simple model in which molecule entry is driven by direct exchange. The CARS microscopic study of the effect of pulse duration confirms that pulses, in the ms time range, induce loss of lipids and membrane deformations facing the electrodes.

We discuss experimental observations of DNA uptake induced by electropermeabilisation. First we describe how experiments on giant unilamelar vesicles can be used to understand the effect of electric fields on lipid membranes and the associated transfer of DNA across the plasma membrane. We then discuss how DNA interacts with electropermeabilised cells in culture and how gene expression is correlated to observations of interactions between DNA and the cell membrane. Finally we discuss how DNA uptake occurs and can be optimized for the most medically relevant case of tissues.

This review presents an overview of the effects of electric fields on giant unilamellar vesicles. The application of electrical fields leads to three basic phenomena: shape changes, membrane breakdown, and uptake of molecules. We describe how some of these observations can be used to measure a variety of physical properties of lipid membranes or to advance our understanding of the phenomena of electropermeabilization. We also present results on how electropermeabilization and other liposome responses to applied fields are affected by lipid composition and by the presence of molecules of therapeutic interest in the surrounding solution.