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Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery
Published on: August 7, 2014
Control of the release of freely diffusing molecules in single-cell electroporation
Aparna Agarwal1, Manyan Wang, Jessica Olofsson
1Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.
Analytical Chemistry
|September 8, 2009
Summary
Single-cell electroporation outcomes depend on cell size and capillary tip distance. Optimizing this distance improves fluorescence control and reduces experimental variability in adherent cells.
Area of Science:
- Biotechnology
- Cell Biology
- Bioengineering
Background:
- Single-cell electroporation is a novel method for creating temporary pores in adherent cells.
- Traditional models like the Schwan equation are unsuitable due to inconsistent cell shapes and non-uniform electric fields.
Purpose of the Study:
- To identify key experimental and cellular parameters influencing single-cell electroporation.
- To develop a predictive model for controlling electroporation outcomes.
Main Methods:
- Adherent A549 cells were loaded with the thiol-reactive dye Thioglo-1.
- Intracellular fluorescence decay was monitored post-electroporation.
- A model relating fluorescence to cell size and tip-to-cell distance was developed and validated.
Main Results:
- Final intracellular fluorescence after electroporation correlates with cell size and capillary tip-to-cell distance.
- Adjusting tip-to-cell distance based on cell size effectively controls fluorescence.
- This method was successfully applied to A549, DU 145, and PC-3 cell lines.
Conclusions:
- A new relationship was established between cell size, tip-to-cell distance, and electroporation outcome.
- Controlling tip-to-cell distance based on cell size significantly reduces fluorescence variability.
- This finding offers enhanced control for single-cell electroporation applications.
