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Related Experiment Video

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The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection

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The temperature effect during pulse application on cell membrane fluidity and permeabilization.

M Kanduser¹, M Sentjurc, D Miklavcic

¹University of Ljubljana, Faculty of Electrical Engineering, Trzaska 25, SI-1000 Ljubljana, Slovenia. masa.kanduser@fe.uni-lj.si

Bioelectrochemistry (Amsterdam, Netherlands)

|May 27, 2008

Summary

Lowering cell suspension temperature significantly reduces cell membrane permeabilization during electroporation. This effect is most pronounced at higher electric field strengths, impacting membrane fluidity.

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Area of Science:

Biophysics
Cell Biology
Electropermeabilization

Background:

Cell membrane permeabilization is induced by high-intensity electric pulses.
Permeabilization extent depends on pulse parameters, media, and cell properties.
Temperature's role in electropermeabilization and membrane fluidity remains under investigation.

Purpose of the Study:

To investigate the effect of temperature during electric pulse application on cell membrane fluidity and electropermeabilization.
To determine the influence of temperature on V-79 and B16F-1 cell lines' response to electroporation.

Main Methods:

Cell membrane fluidity assessed using electron paramagnetic resonance (EPR).
Cell membrane electropermeabilization evaluated via bleomycin uptake and clonogenic assay.
Cells exposed to trains of rectangular electric pulses (500-900 V/cm, 100 µs duration, 1 Hz frequency) after chilling to 4°C.

Main Results:

Chilling cell suspension from 37°C to 4°C significantly decreased cell membrane electropermeabilization.
The most pronounced differences were observed at 900 V/cm electric pulse amplitude.
Lowering temperature reduced membrane fluidity, increasing order parameters and the proportion of highly ordered domains.

Conclusions:

Cell membrane fluidity and domain structure are key factors influencing electropermeabilization.
Temperature plays a critical role in modulating the extent of cell membrane permeabilization by electric pulses.
Additional factors beyond fluidity and domain structure likely contribute to electropermeabilization outcomes.