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Monitoring Electroporation-Induced Changes in Action Potential Generation in Genetically Engineered Tet-On Spiking HEK cells

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Quantification of propidium iodide delivery using millisecond electric pulses: experiments.

Mohamed M Sadik¹, Jianbo Li, Jerry W Shan

¹Department of Mechanical and Aerospace Engineering, Rutgers, The State University of New Jersey, 98 Brett Road, Piscataway, NJ 08854, USA.

Biochimica Et Biophysica Acta

|January 15, 2013

Summary

Electroporation

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Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

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Published on: August 7, 2014

Area of Science:

Biophysics
Cell Biology
Molecular Delivery

Background:

Electroporation is a key technique for molecular delivery into cells.
Understanding transport mechanisms is crucial for optimizing electroporation efficiency.

Purpose of the Study:

To experimentally investigate and quantify transport mechanisms in electroporation-mediated molecular delivery.
To elucidate the role of extracellular conductivity and electrokinetic phenomena in molecular uptake.

Main Methods:

Time- and space-resolved fluorescence microscopy of propidium iodide (PI) uptake in single 3T3 fibroblasts.
Investigation as a function of extracellular buffer conductivity.
Comparison of experimental results with a compact predictive model.

Main Results:

Molecular uptake (PI fluorescence intensity) showed an inverse correlation with extracellular conductivity during electroporation.
Field-Amplified Sample Stacking (FASS) was identified as a key electrokinetic phenomenon.
Electrophoresis was found to be the dominant transport mechanism over diffusion.

Conclusions:

Experimental and modeling approaches provide quantitative insights into electroporation transport.
The findings offer a method for the quantitative diagnosis of electroporation processes.
Optimizing extracellular conductivity can enhance molecular delivery efficiency.