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Journal of Biomedical Optics • Open Access

Nanosecond pulsed electric field thresholds for nanopore formation in neural cells
Author(s): Caleb C. Roth; Gleb P. Tolstykh; Jason A. Payne; Marjorie A. Kuipers; Gary L. Thompson; Mauris N. DeSilva; Bennett L. Ibey

Paper Abstract

The persistent influx of ions through nanopores created upon cellular exposure to nanosecond pulse electric fields (nsPEF) could be used to modulate neuronal function. One ion, calcium (Ca 2+), is important to action potential firing and regulates many ion channels. However, uncontrolled hyper-excitability of neurons leads to Ca 2+ overload and neurodegeneration. Thus, to prevent unintended consequences of nsPEF-induced neural stimulation, knowledge of optimum exposure parameters is required. We determined the relationship between nsPEF exposure parameters (pulse width and amplitude) and nanopore formation in two cell types: rodent neuroblastoma (NG108) and mouse primary hippocampal neurons (PHN). We identified thresholds for nanoporation using Annexin V and FM1-43, to detect changes in membrane asymmetry, and through Ca 2+ influx using Calcium Green. The ED50 for a single 600 ns pulse, necessary to cause uptake of extracellular Ca 2+ , was 1.76  kV/cm for NG108 and 0.84  kV/cm for PHN. At 16.2  kV/cm , the ED50 for pulse width was 95 ns for both cell lines. Cadmium, a nonspecific Ca 2+ channel blocker, failed to prevent Ca 2+ uptake suggesting that observed influx is likely due to nanoporation. These data demonstrate that moderate amplitude single nsPEF exposures result in rapid Ca 2+ influx that may be capable of controllably modulating neurological function.

Paper Details

Date Published: 26 March 2013
PDF: 11 pages
J. Biomed. Opt. 18(3) 035005 doi: 10.1117/1.JBO.18.3.035005
Published in: Journal of Biomedical Optics Volume 18, Issue 3
Show Author Affiliations
Caleb C. Roth, The Univ. of Texas Health Science Ctr. at San Antonio (United States)
Gleb P. Tolstykh, National Research Council (United States)
Jason A. Payne, Air Force Research Lab. (United States)
Marjorie A. Kuipers, Air Force Research Lab. (United States)
Gary L. Thompson, National Research Council (United States)
Mauris N. DeSilva, General Dynamics Information Technology (United States)
Bennett L. Ibey, Air Force Research Lab. (United States)

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