Re: cloverslipping media for fluorescence

From:Gayle Callis <>

No, exposure to UV light will eventually quench fluroescence.  Have used
Vectashield, messy and coverslips moved, Aquamount works, but FITC and
others will eventually fade.  Others are available, but it is not just the
mounting media that causes quenching of fluorescence aka fading.

We use and prefer Alexa fluorchromes from Molecular Probes. Alexa 488 (FITC
look alike) or 546 (rhodamine look alike) are far superior, resist
quenching better than FITC and others.  Brighter also.  They now have an
array of secondary antibodies, some adsorbed for murine work, etc,
conjugated to these special fluorochromes.  They also work very well for
FACS technics.  We love the Strepavidin-conjugates.  

One researcher uses an older Zeiss halogen UV scope and can look at
phycoeritherin (sp?)aka PE for a long time, up to 30 minutes without
fading, also FITC, TRITC, TexRed.  

Make sure you store fluorochromes in dark, keep diluted antibodies in dark
until application/incubation, incubate in dark (under a black towel, etc)
and keep in dark until read.  Protection from light helps.

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

406 994-6367
404 994-4303 (FAX)

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