Speaker
Description
Electroporation (EP) is the increase of permeability of biological membranes to ions and macromolecules, achieved with intense and short pulsed electric fields (PEFs). Depending on the PEF conditions, cells can either restore the plasma membrane integrity (reversible EP, REP), or die due to extensive, non-repairable damage (irreversible EP, IRE). REP is used in biotechnology and medicine to modify cell properties by introducing membrane‐impermeable substances. IRE is a focal, non-thermal ablative technique currently employed for treating cardiovascular and malignant diseases. Despite the established applications, mechanisms of EP have not been fully elucidated. Among the parameters affecting EP, temperature plays a critical role that should be accounted for depending on the specific application.
In this contribution, 2D and 3D realistic, numerical models of A375 human melanoma cells were reconstructed from confocal microscopy live imaging, to be used for electromagnetic and thermal numerical analyses. A good agreement was obtained between the dimensions of the numerical models and the ones measured by the confocal microscope. In real time experiments, A375 cells were exposed to 16 pulses, 100 μs, 1 Hz, 1200 and 1850 V/cm by a customized electric pulse applicator, and EP was quantified by measuring the uptake of PI over time. The same exposure conditions are currently being applied to study temperature dynamics in A375 cells by using the temperature-sensitive dye Rhodamine B.
The objective is to elucidate the physical determinants of the phenomenon, optimize the experimental design, and support the interpretation of the biological processes behind susceptibility to PEFs.
