Open AccessFlow Cytometric Measurement of Efflux in Candida Species
Author(s) -
Iyer Kali R.,
Robbins Nicole,
Cowen Leah E.
Publication year - 2020
Publication title -
current protocols in microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 35
eISSN - 1934-8533
pISSN - 1934-8525
DOI - 10.1002/cpmc.121
Subject(s) - efflux , nile red , flow cytometry , biology , candida albicans , fluorescence , microbiology and biotechnology , fluorescence microscope , cytometry , chromatography , biochemistry , chemistry , physics , quantum mechanics
A technique to assess the ability of distinct Candida strains to efflux substrates, as well as to compare the effectiveness of efflux inhibitors, is important for analysis of antifungal drug resistance mechanisms and the mode of action of antifungals. We describe a method that measures the ability of Candida 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‐resistant Candida species. © 2020 Wiley Periodicals LLC Basic Protocol 1 : Growth and sample preparation of stained Candida Basic Protocol 2 : Quantitative measurement of fluorescence by flow cytometry Alternate Protocol : Qualitative determination of fluorescence using microscopy