22 February 2017 The influence of medium conductivity on cells exposed to nsPEF
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Abstract
Nanosecond pulsed electric fields (nsPEF) have proven useful for transporting cargo across cell membranes and selectively activating cellular pathways. The chemistry and biophysics governing this cellular response, however, are complex and not well understood. Recent studies have shown that the conductivity of the solution cells are exposed in could play a significant role in plasma membrane permeabilization and, thus, the overall cellular response. Unfortunately, the means of detecting this membrane perturbation has traditionally been limited to analyzing one possible consequence of the exposure – diffusion of molecules across the membrane. This method has led to contradictory results with respect to the relationship between permeabilization and conductivity. Diffusion experiments also suffer from “saturation conditions” making multi-pulse experiments difficult. As a result, this method has been identified as a key stumbling block to understanding the effects of nsPEF exposure. To overcome these limitations, we recently developed a nonlinear optical imaging technique based on second harmonic generation (SHG) that allows us to identify nanoporation in live cells during the pulse in a wide array of conditions. As a result, we are able to explore and fully test whether lower conductivity extracellular solutions could induce more efficient nanoporation. This hypothesis is based on membrane charging and the relative difference between the extracellular solution and the cytoplasm. The experiments also allow us to test the noise floor of our methodology against the effects of ion leakage. The results emphasize that the electric field, not ionic phenomenon, are the driving force behind nsPEF-induced membrane nanoporation.
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Erick K. Moen, Erick K. Moen, Bennett L. Ibey, Bennett L. Ibey, Caleb C. Roth, Caleb C. Roth, Ronald A. Barnes, Ronald A. Barnes, Hope T. Beier, Hope T. Beier, Andrea M. Armani, Andrea M. Armani, } "The influence of medium conductivity on cells exposed to nsPEF", Proc. SPIE 10066, Energy-based Treatment of Tissue and Assessment IX, 100660W (22 February 2017); doi: 10.1117/12.2251193; https://doi.org/10.1117/12.2251193
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