Permeant vital stains for yeasts
brambl at graz.cbs.umn.edu
Fri Apr 1 11:42:18 EST 1994
In article <01HAMMGE7PIQ005655 at GW.AGR.CA> BISSETTJ at NCCCOT2.AGR.CA (John
> An industry client has an urgent need of a permeant vital
> stain for Saccharomyces cerevisiae and other yeasts. Their
> automated imaging system cannot reliably distinguish yeasts
> on grid membrane filters from dust and other
> autofluorescing contaminants. The stain should not be
> toxic to yeasts. A fluorescent brightener should have an
> emission range other than 415-505 nm, the range of
> autofluorescence in this application.
> Has anyone knowledge of vital stains/optical brighteners
> for this application - or another solution?
> Please address replies to: BISSETTJ at NCCCOT2.AGR.CA
> John Bissett
An excellent source for a large variety of dyes is a company called
Molecular Probes (Eugene, Oregon). A group of dyes you should consider
are the potentiometric cyanine dyes, some of which report the energization
of the mitochondrial membrane and which are easily measured with
fluorescence microscopy. Others may report pH of the cytosol. One of
these which works really well for fungi is DiOC7(3), the full name of
which either Molecular Probes or I can provide, if necessary. If
autofluorescence remains a problem with your measurements, this could be
subtracted by treatment of the yeast cell samples with CCCP or DCCD.
Hope this helps.
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