Rapid viability assessment of yeast cells using vital staining with 2-NBDG, a fluorescent derivative of glucose.
A fluorescent glucose analogue, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG), which had been developed previously for the analysis of glucose uptake activity by living cells, was investigated to evaluate its applicability for assaying the viability of yeasts. Fluorescence intensities of the yeast population were measured by fluorescence spectrophotometry upon exposure to antifungal agents after staining with 2-NBDG and were compared to the number of colony forming units (CFU). A good correlation was obtained between the yeast viability, determined by the CFU, and the accumulation of 2-NBDG by yeast cells (correlation constant: r=0.98). Susceptibility testing of amphotericin B and miconazole against yeast strains by plate count and 2-NBDG fluorescence method yielded corresponding results. In conclusion, we found that staining with 2-NBDG is a rapid and sensitive method for the assessment of yeast cell viability.
Oh
Ki-Bong
KB
Natural Products Research Institute, Seoul National University, Jongro, South Korea. ohkibong@snu.ac.kr
Matsuoka
Hideaki
H
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000378:metabolism*; D000666:Amphotericin B / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology*; D015169:Colony Count, Microbial; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000378:metabolism*; D005456:Fluorescent Dyes; D005947:Glucose / Q000378:metabolism; D008825:Miconazole / Q000494:pharmacology; D013194:Staining and Labeling; D013997:Time Factors; D015003:Yeasts / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*
D000935:Antifungal Agents; D005456:Fluorescent Dyes; D008825:Miconazole; D000666:Amphotericin B; D003847:Deoxyglucose; D009327:4-Chloro-7-nitrobenzofurazan; D005947:Glucose; C098340:2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose