immunofluorescent staining and paraformaldehyde fixation

From:Gayle Callis <>

It will work.  Don't overfix the cells (monolayer?), I have found 10 min
overkill, often use less, 2 min.  Have done time studies to optimize at 2,
3, 4, 5, etc. and incubated overnight at 4C, in dark.  

This was addressed the other day, we often use Molecular Probes Alexa 488
in place of FITC and Alexa 546 in place of TRITC (rhodamine) for brighter
fluorescence, and they do not quench as easily in UV light.

Histonet archives has many discussions on this, you may want to do a search.


Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
Bozeman MT 59717-3610

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