[Histonet] Long term storage of tissue
renton louise mrs
rentonlf <@t> bru.wits.ac.za
Thu Sep 23 02:06:30 CDT 2004
Hmm, interesting question,
this may not be an answer to your direct query, but from personal expereince I once tried to do PCR on FFPE blocks that were about 50 years old and found that the DNA was largely degraded. There are of course many explanations to this, one being that the formalin used way back then might not have been properly buffered (one favourite technique was to throw a handful of marble chips in the bottom ofthe 10% formalin container, so my assumption remains however that SOME degradation of SOME tissue factors may occur.
-----Original Message-----
From: Robert Krug <rkrug <@t> sial.com>
To: histonet <@t> lists.utsouthwestern.edu
Date: Wed, 22 Sep 2004 13:57:28 -0500
Subject: [Histonet] Long term storage of tissue
I have a question concerning the long term storage of tissue which is not
addressed in any of the histology textbooks I have on hand. After many
hours of searching, I was unable to find an answer through a websearch.
Can tissue be preserved in 10% neutral buffered formalin for decades
without changing the 10% NBF (provided the container is sealed and does
not leak)?
A former coworker of mine once worked at the CDC and she stated the CDC
used 70% alcohol for the long term storage of tissue. A review of the
Internet shows that the Australian Museum Fish site states they have fixed
in "formaldehyde and then stored in alcohol" for the last 80 years. This
site claims to have thousands of specimens on hand. Another site I
found recommended storage in formalin as a way to keep the original color
of the tissue and recommended against long term storage of tissue in
alcohol for various reasons.
The unused 10% neutral buffered formalin solution does decompose over time
and is typically assigned an expiration period. Once the tissue has been
fixed in 10% neutral buffered formalin, is the reaction stable, meaning
the bonds are stable and no decomposition will occur? I was always told
the formalin solution degrades with time and may form formic acid and
other by products. Given enough time the remaining formalin in solution
might form formic acid, drop out of solution as paraformaldehyde or
escape as formaldehyde gas. Another coworker feels that once fixation
has occurred, the fixation is permanent and there is no need to change the
solution. I realize the fixation bonds can be broken through washing in
running water and with select chemicals. But given a sealed container,
will the bonds remain permanent and the tissue free of decomposition or
fungal growth?
Any jounal articles or textbook references you have to support your view
point would be greatly appreciated.
Many thanks
Bob Krug
St John, Missouri
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Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
.......so what IS the speed of dark?
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