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Mary,

we made good experiences with nail hardener (non-fluorescent; check
performance / water resistance by submersing test sample in water after
drying of the hardener). Brands are changing continuously, so it is wise to
check yourself which one to use.

Glass spacers made from cover slips keep the distance between slide and
cover slip and thus help to retain the vectashield under the cover slip. It
is important to remove any embedding solution from the prospective glue
deposit sites on the slide (we use Q-Tips or filter paper). Cover slip
edges are fixed first. After some minutes, the seal can be completed.

No bad influence of nail hardener on fluorescent protein fluorescence
noticed yet, but we did not perform a detailed check regarding this issue.

Hope this helps,

Guenter

At 15:51 23.11.04, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>I've heard varying opinions regarding the use of sealants for slides
>mounted in fluorescence friendly media such as Vectashield.  We are
>looking at eGFP cells with immuno label for cell type markers visualize
>with either Cy3 or Cy5.  Is it true that nail polish is problematic for
>eGFP preps and if rubber cement is a better sealing option, any
>suggestions on how to apply it best? We've tried applicator sticks or
>pastuer pipets with more mess than nail polish.  Any secrets that can be
>shared?
>
>Thanks!
>Mary Ellen
>
>Mary Ellen Pease, M.S.
>Laboratory Manager
>Glaucoma Research Lab & Microscopy and Imaging Core Facility
>Wilmer Eye Institute, Johns Hopkins Hospital
>175 Woods Research
>600 N. Wolfe Street
>Baltimore, MD 21287
>410-955-3337 (phone/voicemail)
>443-287-2711 (fax)
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------------------------------------------
Dr. Guenter Giese
Light Microscopy Facility
Dept. of Biomedical Optics, MPI fuer Medizinische Forschung
Jahnstr. 29, D-69120 Heidelberg, Germany
Phone (+49) 6221-486-360 (Fax: -325)
e-mail: [log in to unmask]
http://www.mpimf-heidelberg.mpg.de/~ggiese/lightmicro