Abstract Determination of the lipid bilayer breakdown voltage by means of linear rising signal

Electroporation is characterized by formation of structural changes within the cell membrane, which are caused by the presence of electrical field. It is believed that bpores ” are mostly formed in lipid bilayer structure; if so, planar lipid bilayer represents a suitable model for experimental and theoretical studies of cell membrane electroporation. The breakdown voltage of the lipid bilayer is usually determined by repeatedly applying a rectangular voltage pulse. The amplitude of the voltage pulse is incremented in small steps until the breakdown of the bilayer is obtained. Using such a protocol each bilayer is exposed to a voltage pulse many times and the number of applied voltage pulses is not known in advance. Such a pre-treatment of the lipid bilayer affects its stability and consequently the breakdown voltage of the lipid bilayer. The aim of this study is to examine an alternative approach for determination of the lipid bilayer breakdown voltage by linear rising voltage signal. Different slopes of linear rising signal have been used in our experiments (POPC lipids; folding method for forming in the salt solution of 100 mM KCl). The breakdown voltage depends on the slope of the linear rising signal. Results show that gently sloping voltage signal electroporates the lipid bilayer at a lower voltage then steep voltage signal. Linear rising signal with gentle slope can be considered as having longer pre-treatment of the lipid bilayer; thus, the corresponding breakdown voltage is lower. With decreasing the slope of linear rising signal, minimal breakdown voltage for specific lipid bilayer can be determined. Based on our results, we suggest determination of lipid bilayer breakdown voltage by linear rising signal. Better reproducibility and lower scattering are obtained due to the fact that each bilayer is exposed to electroporation treatment only once. Moreover, minimal breakdown voltage for specific lipid bilayer can be determined

Responses of plant cells and tissues to pulsed electric field treatments

Cell membrane electroporation/permeabilization may be achieved without affecting cell viability through strict control of the electric pulse parameters. This process is referred to as reversible permeabilization. Even if the cells survive the electric field treatment, they are subjected to stress due to the opening of pores and the struggle of the cells to recover their normal functionality. Very little is known about what actually occurs in the cell and its membranes at the molecular level upon reversible electroporation, and the physiological responses to pulsed electric field (PEF)-induced stress are still largely unknown. This chapter explores the current state of the art on the influence of the complexity of plant tissues on electroporation. Focusing on reversible electroporation, metabolic responses of plant cells and tissues induced by PEF application are also reviewed. One of the first challenges when electroporating plant tissue is their heterogeneous structures where cells vary in shape, size, and cell wall structure. This heterogeneity influences the effect of different electric fields protocols aiming at permeabilizing all cells in the tissue. Once cells are reversibly permeabilized, physiological responses to PEF-induced stress include the production of reactive oxygen species, mobilization of stored energy, activation of stress-related genes, and the production of secondary metabolites. The application of reversible PEF has also been shown to barley seed germination as well as to increase the strength of the cell wall in potatoes and, in consequence, their textural properties. This chapter finishes by revising the effect of reversible PEF on protoplasts (plant cells where the cell walls have been removed) and, in consequence, on the regeneration of new plants. Overall, reports on reversible permeabilization of plant cells and tissues are not common in the literature; however, they have laid the foundation for a fascinating area of research and technological innovation