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Hi Michelle,

in our hands everything that involves heating like paraffin embedding
and polymerizing Epon embedded specimens will destroy GFP fluorescence.
For light microscopy, we have good results with glycolmethycrylate
resins that can be cured in the cold. I guess the same would be true
with EM resins that can be polymerized in the cold. We have not yet
tested that.

Good luck,

Volker

Michelle Peckham wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Dear all
>
>A colleague of ours has asked for  some advice. He has some temporal
>bones (including cochlea and the vestibular apparatus), which have stem
>cells marked with GFP. The specimens have been fixed in PFA and have
>subsequently been demineralised in EDTA. Instead of cryostat sectioning
>would it be possible to embed in plastic(Epon) or paraffin or is the
>dehydration process incompatible with the GFP? He has not previously
>done any work with GFP and the specimens are rather unique so he does
>not want to set up a new and untried protocol.
>
>If anyone has any advice, please let me know.
>
>Thanks
>
>Michelle
>Dr Michelle Peckham
>School of Biomedical Sciences
>Worsley Building
>University of Leeds
>Leeds
>LS2 9JT
>UK
>
>telephone: 0113 343 4348
>fax:       0113 343 4228
>website:   http://www.cell-motility.com
>
>

--
--------------------------------------
Dr. Volker Brinkmann
Mikroskopie
MPI für Infektionsbiologie
Schumannstr. 21-22
10117 Berlin

Tel. +49 (0) 30 28460-  318
Fax                     301
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