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For a long time, estimations and actual measurements of the electric plasma membrane potential (PMP) in whole yeast cells have been the subject of studies by several groups without reliable results. The conditions in the measurements, as well as precautions required to perform them, are described here. Essentially, two approaches using different dyes are reviewed: (a) qualitative estimations by following fluorescence changes under different energization conditions and (b) measurements of the PMP by the accumulation of dyes. An analysis is presented regarding the conditions recommended to obtain more consistent results when following the fluorescence changes. Also, measurements of accumulation of different dyes, and the necessary conditions to perform them, are analyzed. In particular, using acridine yellow appears to be a trustworthy method, with few reserves, both to follow in real time the qualitative changes of the PMP by fluorescence changes and to assess actual PMP values by measuring the accumulation of the dye.

The Plasma Membrane Electric Potential in Yeast: Probes, Results, Problems, and Solutions: A New Application of an Old Dye?