Real-time imaging of membrane potentials during exposure to nanosecond pulsed electric fields

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

Research output: Contribution to journalConference article

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.

Original languageEnglish
Article number48
Pages (from-to)270-276
Number of pages7
JournalProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume5692
DOIs
StatePublished - 2005
EventAdvanced Biomedical and Clinical Diagnostic Systems III - San Jose, CA, United States
Duration: Jan 23 2005Jan 26 2005

Keywords

  • Membrane potential
  • Nanosecond pulsed electric fields
  • Real-time fluorescence imaging

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    Kolb, J. F., Frey, W., White, J. A., Price, R. O., Blackmore, P. F., Beebe, S. J., Joshi, R. P., & Schoenbach, K. H. (2005). Real-time imaging of membrane potentials during exposure to nanosecond pulsed electric fields. Progress in Biomedical Optics and Imaging - Proceedings of SPIE, 5692, 270-276. [48]. https://doi.org/10.1117/12.591313