Abstract
Hyphae of the oomycete Saprolegnia ferax were stained with diverse neutral, anionic, cationic, and phospholipid analog fluorescent membrane probes. Apparently only probes smaller than 1000 Da enter the living hyphae but these permitted near normal growth rates, thus demonstrating their use as vital fungal stains. Collectively the penetrant probes stained all identifiable membranes, but individually none faithfully revealed the total cellular membranes. However, several probes produced a diffuse sub-apical concentration of stain which could not be correlated with any specific membrane type but which may indicate the highest concentration of total cellular membranes. The neutral probes were unique in staining unidentified spherical refractile vesicles. Nile Red was the only probe to show significant staining in the extreme apex where wall vesicles are abundant. DiOC 6 (3) at low concentration was an excellent mitochondria stain as previously reported but at the higher concentrations used in other cells as an endoplasmic reticulum selective stain it was toxic and caused mitochondrial deformation. NDB hexanoic ceramide failed to specifically stain Golgi bodies as previously described in mammalian cells. Our results show that considerable care must be exercised in extrapolating the use of these probes from one cell type to another and in interpreting their staining patterns in new cells.
Published Version
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