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Related Experiment Videos

Nanoelectropulse-driven membrane perturbation and small molecule permeabilization.

P Thomas Vernier1, Yinghua Sun, Martin A Gundersen

  • 1Department of Electrical Engineering-Electrophysics, Viterbi School of Engineering, University of Southern California, Los Angeles, CA 90089-0271, USA. vernier@mosis.com

BMC Cell Biology
|October 21, 2006
PubMed
Summary

Short, high-voltage electric pulses as brief as 3 nanoseconds can alter cell membranes and cause pore formation. This supports the theory that electric field charging drives nanosecond membrane poration and phosphatidylserine (PS) externalization.

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Area of Science:

  • Cellular Biophysics
  • Electroporation Studies
  • Membrane Dynamics

Background:

  • Nanosecond, megavolt-per-meter pulsed electric fields are known to disrupt cell membranes, leading to calcium release and apoptosis.
  • Previous research linked phospholipid rearrangement to nanoelectropulse exposure, suggesting transmembrane potential drives phosphatidylserine (PS) externalization.

Purpose of the Study:

  • To investigate cellular responses to electric pulses with durations as short as 3 nanoseconds (ns).
  • To differentiate the effects of unipolar versus bipolar pulses on membrane phospholipids.
  • To test the membrane charging hypothesis regarding pulse duration and electric field strength.

Main Methods:

  • Exposure of cells to nanosecond pulsed electric fields (3 ns to 30 ns).

Related Experiment Videos

  • Tracking phospholipid order changes using FM1-43 fluorescence.
  • Monitoring influx of fluorescent dyes YO-PRO-1 and propidium iodide to assess membrane permeabilization.

    • Main Results:

    • Pulses as short as 3 ns were sufficient to induce cellular responses similar to longer pulses.
    • Bipolar pulses redistributed phospholipids at both anode and cathode poles, unlike unipolar pulses.
    • Shorter pulse trains required higher fields for comparable phospholipid scrambling, supporting the membrane charging hypothesis.
    • Sufficiently high numbers of pulses induced YO-PRO-1 influx, indicating membrane poration, with propidium iodide entry at higher pulse counts.

    Conclusions:

    • Electric pulses as short as 3 ns can alter plasma membrane structure and permeabilize cells.
    • Dose responses to pulses (3-30 ns) support a field-driven charging mechanism for nanosecond pore formation.
    • Anionic phospholipid PS migrates electrophoretically to the external membrane face during nanosecond electroporation.