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1 April 2005 Real-time imaging of membrane potentials during exposure to nanosecond pulsed electric fields
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Abstract
The charging of mammalian cell plasma membranes in response to ultrashort pulsed electric fields of 60 ns and field strengths up to 100 kV/cm was investigated. Membranes of Jurkat cells were stained with a potential-sensitive dye, Annine-6 and placed in a microreactor mounted on an inverted fluorescence microscope. Images of changes in the fluorescence intensity during the exposure were recorded with a high-sensitivity CCD-camera. A temporal resolution of 5 ns was achieved by illuminating the cells with a 5 ns laser pulse from a dye-laser. The laser pulse was synchronized with the high voltage pulse to record images at specific times before, during and after exposure to the electric field. When exposing Jurkat cells to a 60 ns, 100 kV/cm pulse, each hemisphere of the plasma membrane (as oriented with respect to the electrodes) responded uniquely to the applied field. From these observations it is possible to draw conclusions on the charging time of the membrane, maximum transmembrane voltages and the onset of poration.
© (2005) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Juergen F. Kolb, Wolfgang Frey, Jody A. White, R. O. Price, Peter F. Blackmore, Stephen J. Beebe, Ravindra P. Joshi, and Karl H. Schoenbach "Real-time imaging of membrane potentials during exposure to nanosecond pulsed electric fields", Proc. SPIE 5692, Advanced Biomedical and Clinical Diagnostic Systems III, (1 April 2005); https://doi.org/10.1117/12.591313
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