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We describe a method that measures the ability of\n                    <jats:italic>Candida<\/jats:italic>\n                    species to extrude the fluorescent dye Nile red as an output for efflux activity. This involves exposing cells to Nile red and using flow cytometry to quantify cellular fluorescence, enabling numerous samples to be processed in a limited time frame. This protocol provides a simple, yet effective method for quantifying efflux in drug\u2010resistant\n                    <jats:italic>Candida<\/jats:italic>\n                    species. \u00a9 2020 Wiley Periodicals LLC\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Growth and sample preparation of stained\n                    <jats:italic>Candida<\/jats:italic>\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Quantitative measurement of fluorescence by flow cytometry\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol<\/jats:bold>\n                    : Qualitative determination of fluorescence using microscopy\n                  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