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The potential of the plasma membrane (&#916;&#968;) regulates the electrochemical potential between the outer and inner sides of cell membranes. The opportunistic fungal pathogen, <i>Candida albicans,</i> regulates the membrane potential in response to environmental conditions, as well as the physiological state of the cell. Here we demonstrate a new method for detection of cell membrane depolarization/permeabilization in <i>C. albicans</i> using the potentiometric zwitterionic dye di-4-ANEPPS. Di-4-ANEPPS measures the changes in the cell &#916;&#968; depending on the phases of growth and external factors regulating &#916;&#968;, such as potassium or calcium chlorides, amiodarone or DM-11 (inhibitor of H⁺-ATPase). We also demonstrated that di-4-ANEPPS is a good tool for fast measurement of the influence of amphipathic compounds on &#916;&#968;.