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Dear Damir--

On 5/3/2011 12:58 PM, Damir Sudar wrote:

> I would really appreciate some good suggestions how to keep
> immunofluorescently labeled slides of mammalian cells good for long
> periods (3-5 months). The fluors are typically Alexa dyes or similar. Is
> freezing at -20 or -80 a good idea? Any preparation tricks that keep the
> fluorescence and the localization intact?

We stain tissue sections but I think our method should work for cells as 
well.  We stain with Cy2, Cy3, rhodamine and Cy5-labeled secondaries 
(and in the old days, Bodipy) and/or DNA dyes--but NOT fluorescein. 
After staining, we dehydrate the tissue in graded alcohols, clear in 
xylene and mount with DPX.  The DNA dyes tend to photobleach but 
photobleaching of the secondaries is acceptably low for our purposes.

I have fluorescently stained sections that I processed that way almost 
20 years ago and have kept on my desk top in the intervening time, that 
I still use for teaching purposes.  I like having a solid mounting 
medium that doesn't require cold-storage.

Good luck!

Martin Wessendorf


-- 
Martin Wessendorf, Ph.D.                   office: (612) 626-0145
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