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Related Experiment Videos

Characterization of single-cell electroporation by using patch-clamp and fluorescence microscopy.

F Ryttsén1, C Farre, C Brennan

  • 1Department of Chemistry, Göteborg University, Göteborg SE-412 96, Sweden.

Biophysical Journal
|October 12, 2000
PubMed
Summary

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Electroporation of NG108-15 cells using carbon-fiber microelectrodes requires approximately 250 mV for dielectric breakdown. Cell permeabilization depends on analyte concentration and membrane potential, with minimal effects beyond 30 micrometers.

Area of Science:

  • Cellular Electrophysiology
  • Biophysical Characterization
  • Nanotechnology in Biology

Background:

  • Electroporation is a key technique for cell membrane permeabilization.
  • Understanding the precise electrical parameters for electroporation is crucial for controlled cellular manipulation.
  • NG108-15 cells are a relevant model for neuronal studies.

Purpose of the Study:

  • To characterize the electroporation of single NG108-15 cells using carbon-fiber microelectrodes.
  • To determine the electric field strengths and transmembrane potentials required for pore formation.
  • To investigate the kinetics of pore formation and analyte entry.

Main Methods:

  • Patch-clamp recordings to measure transmembrane currents.
  • Fluorescence microscopy to visualize pore formation and analyte uptake.

Related Experiment Videos

  • Controlled application of electrical pulses using carbon-fiber microelectrodes.
  • Optimization of electrode-cell interface to minimize capacitive effects.

    • Main Results:

    • The threshold transmembrane potential for dielectric breakdown was approximately 250 mV for NG108-15 cells with 1-ms pulses.
    • Electroporation pulse preceded pore formation, with analyte entry influenced by concentration and membrane potential.
    • Cells beyond approximately 30 micrometers from the microelectrodes were not permeabilized.
    • Kinetics of pore opening, closing, and open times were investigated.

    Conclusions:

    • Precise control over electroporation parameters, including electrode angle and electrical field strength, is essential.
    • The study provides quantitative data on the electrical thresholds for NG108-15 cell electroporation.
    • This research contributes to the development of targeted and efficient cell permeabilization techniques.