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Measuring the Induced Membrane Voltage with Di-8-ANEPPS
Published on: November 19, 2009
Induced transmembrane voltage and its correlation with electroporation-mediated molecular transport
Tadej Kotnik1, Gorazd Pucihar, Damijan Miklavcic
1Department of Biomedical Engineering, Faculty of Electrical Engineering, University of Ljubljana, Trzaska 25, 1000, Ljubljana, Slovenia.
Abstract:
Exposure of a cell to an electric field results in inducement of a voltage across its membrane (induced transmembrane voltage, DeltaPsi (m)) and, for sufficiently strong fields, in a transient increase of membrane permeability (electroporation). We review the analytical, numerical and experimental methods for determination of DeltaPsi (m) and a method for monitoring of transmembrane transport. We then combine these methods to investigate the correlation between DeltaPsi (m) and molecular transport through an electroporated membrane for isolated cells of regular and irregular shapes, for cells in dense suspensions as well as for cells in monolayer clusters. Our experiments on isolated cells of both regular and irregular shapes confirm the theoretical prediction that the highest absolute values of DeltaPsi (m) are found in the membrane regions facing the electrodes and that electroporation-mediated transport is confined to these same regions. For cells in clusters, the location of transport regions implies that, at the field strengths sufficient for electroporation, the cells behave as electrically insulated (i.e., as individual) cells. In contrast, with substantially weaker, nonelectroporating fields, potentiometric measurements show that the cells in these same clusters behave as electrically interconnected cells (i.e., as one large cell). These results suggest that sufficiently high electric fields affect the intercellular pathways and thus alter the electric behavior of the cells with respect to their normal physiological state.
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