Electrofusion effect of induced transmembrane potential under a live - Cell microscopy system

Abstract

Pulsed electric field (PEF) application development substantially affected the development of research process including controlling cell functions by using pulses of electrical fields to create pores through a cellular membrane causes cell lysis and apoptosis commonly known as electroporation or electrofusion. Here we demonstrate the influence of the PEF on the threshold area (TA) of human cervical cancer cells (HeLa) membrane. The electric field for PEF is 2.7kV/cm while the pulse length and the number of pulses were fixed at 30µs and 1, respectively. While the cultured Hela cells are placed in 9 mm-platinum electrode gap for allowing real time observation of membrane permeability changes and cellular physiology. In order to initiate higher cell viability rate, high transfection efficiency, lower sample contamination and smaller Joule heating effect the modification of chamber need to be done which can be controlled by pH scale, temperature and humidity. The experiment using high pulsed electrical field with single pulses shows the perfusion area of cell membrane was increasing gradually under live-cell microscopy system, and is settled with time lapsed technique within 30 minute. We found that the intensity profile of cells membrane decrease gradually to 82%. This situation will not change until the whole of these cells merge and form a condition called hybrid cells. was affected when exposed to high voltage pulse electric field. The dependence of the electrofusion area on the HeLa cell membrane might be associated with the electrical impedance of the plasma membrane that begins to fluctuate after the application of a certain level of PEF.