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Plasma Membrane Voltage Changes during Nanosecond Pulsed Electric Field Exposure

W. Frey ^*, J. A. White , R. O. Price , P. F. Blackmore , R. P. Joshi , R. Nuccitelli , S. J. Beebe ^¶, K. H. Schoenbach   and J. F. Kolb 

^* Forschungszentrum Karlsruhe, IHM, D-76344, Eggenstein-Leopoldshafen, Germany; Frank Reidy Research Center for Bioelectrics, and Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, Virginia; and Department of Physiological Sciences, and ^¶ Center for Pediatric Research, Eastern Virginia Medical School, Norfolk, Virginia

Correspondence: Address reprint requests to J. F. Kolb, E-mail: jkolb{at}odu.edu.

The change in the membrane potential of Jurkat cells in response to nanosecond pulsed electric fields was studied for pulses with a duration of 60 ns and maximum field strengths of 100 kV/cm (100 V/cell diameter). Membranes of Jurkat cells were stained with a fast voltage-sensitive dye, ANNINE-6, which has a subnanosecond voltage response time. A temporal resolution of 5 ns was achieved by the excitation of this dye with a tunable laser pulse. The laser pulse was synchronized with the applied electric field to record images at times before, during, and after exposure. When exposing the Jurkat cells to a pulse, the voltage across the membrane at the anodic pole of the cell reached values of 1.6 V after 15 ns, almost twice the voltage level generally required for electroporation. Voltages across the membrane on the side facing the cathode reached values of only 0.6 V in the same time period, indicating a strong asymmetry in conduction mechanisms in the membranes of the two opposite cell hemispheres. This small voltage drop of 0.6–1.6 V across the plasma membrane demonstrates that nearly the entire imposed electric field of 10 V/µm penetrates into the interior of the cell and every organelle.

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