Dynamics of Cell Membrane Lesions Under the Electrical Stress
Document Type
Abstract
Publication Date
11-7-2024
DOI
10.25776/1wnw-fv12
Abstract
Cell electropermeabilization, induced by electric fields (EF), is a well-known phenomenon; however, the small and dynamic nature of membrane lesions complicates direct visualization. This limitation has led to reliance on indirect methods, including molecular dynamics simulations and artificial models, though many aspects, such as long-term membrane permeabilization and electropore structure, remain elusive. In this study, we introduce a novel approach to detect and monitor individual electropores in live HEK293 cells. By combining whole-cell patch clamp on cells adhered to an indium tin oxide (ITO) surface with total internal reflection fluorescence (TIRF) imaging, we visualize focal Ca2+-permeable lesions indicative of electropores. EF application between a patch pipette and the ITO surface polarized the plasma membrane, inducing electroporation marked by a step-wise current increase. TIRF time-lapse imaging enabled spatial and temporal tracking of lesions, revealing both transient and persistent electropores, as well as low-threshold zonal permeabilization at transmembrane potentials as low as -100 mV. This is the first direct visualization of individual electropores in live cells, providing a valuable method for probing electropermeabilization mechanisms and laying a foundation for future research on nanosecond EF pulse electroporation and electropore structure.
Repository Citation
Silkunas, Mantas; Silkuniene, Giedre; and Pakhomov, Andrei G., "Dynamics of Cell Membrane Lesions Under the Electrical Stress" (2024). 2024 Frank Reidy Research Center for Bioelectrics Retreat. 4.
https://digitalcommons.odu.edu/bioelectrics-2024retreat/4
