Optimization of irreversible electroporation technique for the study of pore formation in lipid membranes of giant unilamellar vesicles

Abstract

Irreversible electroporation (IRE) is a minimally invasive and non-thermal technique in which an ultra-short pulse with high electric field has been found to be successful for ablation of certain tumor and cancer cells. As a mimic of biomembranes of cells, lipid membranes of giant unilamellar vesicles (GUVs), composed of dioleoylphosphatidylglycerol (DOPG) and dioleoylphosphatidylcholine (DOPC), with diameters equal or greater than 10 μm is currently used. In this regard, at first a microcontroller-based IRE technique is developed indigenously where a special purpose power supply is used to construct high voltage generator and a MOSFET (N-Channel Power MOSFET, SCILLC) based switching circuit is designed for generating square pulses with very high energy as required for electroporation. Then investigated the electro- deformation and pore formation of GUVs at constant tension induced by IRE signal. It is also observed the membrane fusion of GUVs induced by the IRE signal. The constant tension is induced on the GUVs at electric field strength 340V/cm with pulse width 200 µs. The pore formation in GUVs is increased with the increase of constant tension which supported the results of the mechanical tension-induced pore formation. The pore formation in the lipid membranes is explained by the classical theory of pore formation. These results provide important information for the mechanism of IRE-induced pore formation in biomembranes and lipid membranes. Therefore, optimization of various parameters of IRE technique is necessary for the study of pore formation in lipid membranes of GUVs.