GFP and Nail Polish

brett brett at BORCIM.WUSTL.EDU
Thu Mar 5 09:54:44 EST 1998


>To be able to do 3D data collection on our GFP-labeled cells we need to fix
>the coverslip somehow on the slide. Normally nailpolish is used to do this.
>However all those that we've tried decreased the fluorescence of the GFP. Has
>anybody an idea what to use instead?
>
>Thanks in advance for any hints
>
>Steffen Dietzel

You didn't mention if you were working with live or fixed cells. For live
cells, I make a little channel of grease on a clean slide, a little less than
one coverslip in width (in this case, larger coverslips are better - I use
22x40mm rectangles). Plop this down on a PBS-drenched coverslip to make a
"cell sandwich" and seal top and bottom edges by brushing molten paraffin
on with a cotton swab.  Dry the exposed glass surfaces and take a look.
For fixed cells, I fix in 4% paraformaldehyde for 10', rinse in PBS, and
mount in a self-hardening medium containing anti-fade (For example, Molecular
Probes' ProLong reagent (no affiliation)). Since it hardens, there is no need
to use nail polish.






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