Re: Tissue storage

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From:vandeplas@aurion.nl (Peter van de Plas)
To:Histonet@Pathology.swmed.edu
Reply-To:
Date:Tue, 16 Mar 1999 03:50:46 +0200
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Ian Montgomery wrote:
 My group will be fixing small arteries at phyiological pressure
then staining with nuclear dyes for confocal studies. After confocal
microscopy some of the arteries will be processed for routine histology and
ICC (endothelium, smooth muscle, elastin and collagen) while others will
remain in a suitable fluid, just in case we need to repeat the confocal
microscopy. In the past, the arteries were left in formalin as ICC was not
required, but now it might be. Any suggestions as to a long term storage
solution , bearing in mind that it might be at least 6 months or more
before any decisions are made regarding further use.
        I'm dealing with a bunch of Physiologists and Pharmacologists who
will decide after 3-4 months that a particular specimen demonstrated a
result which merits further study. I'll be led in, then asked to perform
the necessary miracle be it histology, ICC or E.M. "Ah but, it was sitting
in formalin for 6 months, it's nae use". "Why not, and why didn't you tell
us earlier".
------------------------------
Dear Ian,
The following method works on cell cultures and small tissue blocks for EM:
After fixation with aldehyde specimens are washed in PBS and treated with
0.1% NaBH4 in PBS for 15 minutes. Specimens are repeatedly washed in PBS
and this buffer is also used for storage. The H2 gas that is formed has no
effect on morphology of the above mentioned specimens. Another possibility
to prevent defixation of your tissue might be the use of a low
concentration of formaldehyde inbuffer.
Hope this is of help,
Peter

========================================
Peter van de Plas
AURION
Costerweg 5
6702 AA Wageningen
The Netherlands
phone: (31)-317-497676
fax: (31)-317-415955
http://www.aurion.nl





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