23 February 2013 Measurement of changes in plasma membrane phospholipid polarization following nanosecond pulsed electric field exposure
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Abstract
The exposure of nanosecond pulsed electric fields (nsPEF) to living cells has been shown to create nanopores in the plasma membranes. These nanopores allow the passage of small ions but exclude the transport of larger molecules such as Propidium ions, with permeabilization persisting for many minutes. To characterize these nanopores and the effect of temperature of the formation and resealing of these pores, we have chosen to use 6-Propionyl-2-(N,NDimethylamo) Naphthalene (PRODAN) as an indicator of membrane organization. PRODAN is a fluorescent dye with a large excited-state dipole moment that displays extensive solvent polarity-dependent fluorescent shifts. By monitoring this shift in fluorescence spectrum, disruption of the membrane after an electric exposure is observed as an immediate increase in the membrane fluidity, likely indicating poration of the membrane. High-speed imaging results indicate that a change in membrane organization occurs instantly (<5 ms), with longer pulse widths having a more dramatic effect on the membrane. This instantaneous membrane disruption was shown to recover within 500 ms.
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Samantha Franklin, Samantha Franklin, Bennett L. Ibey, Bennett L. Ibey, Kelly Nash, Kelly Nash, Hope T. Beier, Hope T. Beier, } "Measurement of changes in plasma membrane phospholipid polarization following nanosecond pulsed electric field exposure", Proc. SPIE 8585, Terahertz and Ultrashort Electromagnetic Pulses for Biomedical Applications, 85850S (23 February 2013); doi: 10.1117/12.2005091; https://doi.org/10.1117/12.2005091
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