[Chlamydomonas] neutral red or other fluorescent vital staining of chlamy

Otti Croze via chlamy%40net.bio.net (by Ottavio.Croze from glasgow.ac.uk)
Mon Oct 11 07:43:15 EST 2010


Dear Chlamy community,

I need to fluorescently stain Chlamydomonas augustae (CCAP 11/51B) cells
for an experiment where we want to image a labelled subpopulation of 
swimming cells flowed in a tube. I want to image the population 
macroscopically, so I don't care what gets stained provided the cells 
are more visible than unstained cells and as normally motile.

I have tried with neutral red but I am failing to stain the cells and
keep them motile. Cells stained at around 100 micro molar of dye and
washed, fluoresce nicely, but they don't swim! Cells stained for smaller
concentrations swim nicely, but are not stained.

Can anyone with experience of neutral red staining of chlamy give me 
some advice? Alternatively can anyone suggest a protocol for staining 
with any other fluorescent vital dye which doesn't affect motility? (I 
am already aware of FDA, but haven't tried it yet, as we don't have the 
right filters).

Thanks in advance for your help!

Otti Croze,
University of Glasgow



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