TY - JOUR
T1 - Saccharomyces cerevisiae samples stained with FUN-1 dye can be stored at -20°C for later observation
AU - Eggleston, M. D.
AU - Marshall, Pamela
PY - 2007/1
Y1 - 2007/1
N2 - FUN-1, a fluorescent vital dye, has been observed to form cylindrical intravacuolar structures within the vacuoles of metabolically active yeast cells. FUN-1 staining, which begins as a diffuse pool of fluorescent cytoplasmic stain, uses an unknown endogenous biochemical processing mechanism to compact and form orange-red cylindrical intravacuolar structures within the cell vacuole. In the clinical setting, FUN-1 is primarily used for identification of fungal infection. FUN-1 is utilized in the laboratory to distinguish between metabolically active and dead fungal cells. Although this stain is useful for distinguishing between live and dead fungal dead cells, few studies have utilized this chemical. This lack of use in the scientific community may be due to the requirement that cells are visualized directly after staining. Thus, it would be of interest to be able to stain cells and store them for later use. Our lab examined the longevity of cylindrical intravacuolar structures in two strains of Saccharomyces cerevisiae stained with FUN-1 and stored at -20°C. We found that cylindrical intravacuolar structures could be reliably observed and imaged utilizing differential interference contrast microscopy and fluorescence microscopy for 21 days. We also observed that cells stained with FUN-1 would resume propagation on yeast extract, peptone, dextrose (YPD) plates after being frozen at -20°C for 21 days. These modifications to the published procedure for FUN-1 dye staining should allow for a more prevalent and less time sensitive use of this important biological tool.
AB - FUN-1, a fluorescent vital dye, has been observed to form cylindrical intravacuolar structures within the vacuoles of metabolically active yeast cells. FUN-1 staining, which begins as a diffuse pool of fluorescent cytoplasmic stain, uses an unknown endogenous biochemical processing mechanism to compact and form orange-red cylindrical intravacuolar structures within the cell vacuole. In the clinical setting, FUN-1 is primarily used for identification of fungal infection. FUN-1 is utilized in the laboratory to distinguish between metabolically active and dead fungal cells. Although this stain is useful for distinguishing between live and dead fungal dead cells, few studies have utilized this chemical. This lack of use in the scientific community may be due to the requirement that cells are visualized directly after staining. Thus, it would be of interest to be able to stain cells and store them for later use. Our lab examined the longevity of cylindrical intravacuolar structures in two strains of Saccharomyces cerevisiae stained with FUN-1 and stored at -20°C. We found that cylindrical intravacuolar structures could be reliably observed and imaged utilizing differential interference contrast microscopy and fluorescence microscopy for 21 days. We also observed that cells stained with FUN-1 would resume propagation on yeast extract, peptone, dextrose (YPD) plates after being frozen at -20°C for 21 days. These modifications to the published procedure for FUN-1 dye staining should allow for a more prevalent and less time sensitive use of this important biological tool.
KW - FUN-1 dye
KW - Saccharomyces cerevisiae
KW - Storage
KW - Yeast
UR - http://www.scopus.com/inward/record.url?scp=33846811001&partnerID=8YFLogxK
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U2 - 10.1111/j.1365-2818.2007.01720.x
DO - 10.1111/j.1365-2818.2007.01720.x
M3 - Article
C2 - 17286700
AN - SCOPUS:33846811001
SN - 0022-2720
VL - 225
SP - 100
EP - 103
JO - Journal of Microscopy
JF - Journal of Microscopy
IS - 1
ER -