Complementary Methods of Processing diS-C3(3) Fluorescence Spectra Used for Monitoring the Plasma Membrane Potential of Yeast: Their Pros and Cons
Abstract
Carbocyanine dye diS-C-3(3) was repeatedly employed in monitoring the plasma membrane potential of yeast and other living cells. Four methods of measuring and evaluating probe fluorescence signal were used in different studies, based on following fluorescence parameters: fluorescence intensity emitted within a certain spectral interval, F(580)/F(560) fluorescence emission ratio, wavelength of emissionspectrum maximum, and the ratio of respective fluorescence intensities corresponding to the diS-C-3(3) bound to cytosolic macromolecules and remaining dissolved in the aqueous cell medium (i.e., unbound, or free).
We have shown that data corresponding to the three latter spectral assessments of diS-C-3(3) accumulation in cells is mutually convertible, which means that their alternative use cannot lead to ambiguities in the interpretation of the results of biological experiments.