[Histonet] NBF fixation and long term storage

[Rene J Buesa]

Patrick:
Under separate cover I am sending an article about NBF fixation and its comparison with other fixatives so you can make an "informed" decision.
René J.

--- On Wed, 9/15/10, Lewis, Patrick <patrick.lewis <@t> seattlechildrens.org> wrote:


From: Lewis, Patrick <patrick.lewis <@t> seattlechildrens.org>
Subject: [Histonet] NBF fixation and long term storage
To: Histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, September 15, 2010, 1:53 PM


I have some questions about Fixing tissues for paraffin embedding when
the ultimate purpose will be looking for viral antigens and cell surface
markers.



We have some monkey tissue that has been in NBF for over a year. and we
are just now processing them and embedding them in to paraffin.



We will be getting additional tissues from monkey necropsies and in an
effort to avoid epitope damage from prolonged exposure to NBF I was
wondering if anyone could recommend a fixation period.  (I.E. no longer
than 24 hours in NBF, 48 hours? ETC.



I was thinking of replacing the NBF (Neutral buffered Formalin) with 70%
Etoh.  And using 70% Etoh as long term storage of tissue parts.  The
ideal situation would be to get the tissues from necropsy the same day
of the necropsy and store the tissues overnight in NBF and then transfer
them to 70% etoh and then process them on the tissue processer after 24
hours in Etoh.  



I'm not sure how many animals I've eventually get and I'm a little
concerned over where we will put all the spare animal parts that I'm
sure I'll accumulate.



We don't want to throw anything away and for some tissues I'll have more
than I can put into a single cassette.

I may end up using more than 1 cassette for a single piece of tissue so
that I have backup cassettes of it.  



Anyone storing research tissues  has a suggestion about the best way to
store them when space is a premium.



Thanks 


Patrick.





Patrick Lewis

Research Associate II-Bench| Infections and Prematurity

Seattle Children's Research Institute

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