Biochemical and Biophysical Research Communications
Nanosecond electric pulses penetrate the nucleus and enhance speckle formation
Section snippets
Materials and methods
Cell culture. Jurkat cells, a human T cell leukemia cell line [American Type Culture Collection (ATCC), Manassas, VA], were grown in RPMI-1640 medium (phenol red) containing 2 mM l-glutamine (ATCC, VA) and supplemented with 10% fetal bovine serum (ATCC, VA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma, St. Louis, MO), and were maintained at 37 °C with 5% CO2. The cells used in our experiments ranged between 30 and 50 passages. We verified that cell viability exceeded 90% daily and prior
Results
Using immune labeling, temporally, and spatially resolved images of nuclear speckles generated by 10 ns pulse-induced stimulation were measured by recording intra-nuclear fluorescence of Jurkat cells caused by the binding of Y12 antibody with FITC to the nuclear speckles. We examined the nuclear speckles before the pulse and at 10, 60, and 180 min post-pulse. Nuclear speckles were detected after pulsing by examining the cellular fluorescence corresponding to the predicted molecular weight of
Discussion
Compared to traditional electroporation, nsPEFs preferentially charge the membranes of subcellular organelles, thereby inducing distinct effects on cellular structure and function that are predominantly intracellular in nature [1], [2], [5], [8]. The resulting delayed plasma membrane permeabilization was likely secondary, arising due to subcellular effects [2] rather than direct electroporation [18], [19]. In previous confocal microscopic real time studies, we compared the nuclear and plasma
Acknowledgments
This work was supported in part by an Air Force Office of Scientific Research (AFOSR) MURI grant on Subcellular Response to Narrow Band and Wide Band Radio Frequency Radiation, administered by Old Dominion University. One author (A.L.G.) was supported by a National Defense Science and Engineering Graduate Fellowship sponsored by the Department of Defense.
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Present address: GE Global Research Center, Niskayuna, NY 12309, USA.