Just curious-what do people consider the time frame for long term ?
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Barry
Rittman
Sent: Monday, December 09, 2013 7:50 AM
To: Orla M Gallagher
Cc: histonet@lists.utsouthwestern.edu; Wineman, Terra
Subject: Re: AW: [Histonet] Bone samples long-term storage in 10% formalin
or 4% paraformaldehyde
hi
I would recommend storage for long term in 70% ethanol. To prevent drying
out we used glycerin in the ethanol, about 20% of the volume.
Barry
On Mon, Dec 9, 2013 at 7:44 AM, Orla M Gallagher
o.m.gallag...@sheffield.ac.uk wrote:
Thanks to everyone for your comments.
I may not have been clear in my question - our researchers don't wish
to decalcify these formalin-fixed bones yet, but rather to store them
for more than a couple of weeks, in case they need to carry out
MicroCT followed by histology later. I'm aware that the formalin or
paraformaldehyde will degrade over time, but I just wondered if anyone
has a protocol for storage without decalcification? I guess transfer
to 70% ethanol is an option but this is also not ideal for longterm
storage, and would need to be removed before decal in EDTA.
All the best,
Orla
On 6 December 2013 16:12, Wineman, Terra terra.wine...@novusint.com
wrote:
I would suggest a different protocol if the tissue will not be
processed for a while. I would say a week in 10%NBF and then
transfer the bones to an EDTA decal solution. The bones will decal
slowly without the affects
of
the formic acid. I am in research and this is what we do with our
bones.
Terra Wineman, HTL (ASCP)CM
Research Biologist
636-926-7476 phone
terra.wine...@novusint.com
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu [mailto:
histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
pru...@ihctech.net
Sent: Thursday, December 05, 2013 2:50 PM
To: gu.l...@gmx.at; 'Orla M Gallagher'
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: AW: [Histonet] Bone samples long-term storage in 10%
formalin
or 4% paraformaldehyde
i would think u are correct in advising formic acid decal and then
processing into paraffin for the best protection of the trap enzyme,
immunoreactivity, etc. A couple of weeks in formalin should be fine.
Paraformaldehyde show be the same as formalin. I do know a way to
restore
the enzyme activity for TRAP that may have been lost so if u need
that
let
me know.
- Original Message - Subject: AW: [Histonet] Bone
samples long-term storage in 10% formalin or 4% paraformaldehyde
From: Gudrun Lang gu.l...@gmx.at
Date: 12/5/13 11:42 am
To: 'Orla M Gallagher' o.m.gallag...@sheffield.ac.uk
Cc: histonet@lists.utsouthwestern.edu
Paraformaldehyd is formaldehyd in solid form. Formalin is the
aequous solution of formaldehyd.
So the main characteristics are the same.
Gudrun Lang
-Urspruuml;ngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von
Orla
M
Gallagher
Gesendet: Donnerstag, 05. Dezember 2013 19:31
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Bone samples long-term storage in 10% formalin
or 4% paraformaldehyde
Dear Histonetters,
What is your opinion on storing bone samples long-term (more than a
couple of weeks) in 10% formalin? As I was taught, best practice
has always been to fix only as long as necessary, depending on the
size of
the
sample, then decalcify and process to wax, and I always stress this
to everyone I advise.
However, research colleagues sometimes wish to do histology on bone
samples that have been stored for months ..or even years! As the
formalin
pH becomes more acidic, there is formalin pigment and the
immunoreactivity
and TRAP enzyme activity is diminished or destroyed during long
fixation,
is there any way of minimising this e.g. has anyone tried regularly
replacing the old formalin with fresh buffered formalin, or storing
formalin-fixed bones in any other medium? I'm also interested in
how
best
to fix in 4% paraformaldehyde and whether the problems are the same
with long-term storage.
Thanks for your comments.
All the best,
Orla
--
**
Ms. Orla Gallagher
Bone Analysis Laboratory
Mellanby Centre for Bone Research
Department of Human Metabolism
D Floor Medical School
University of Sheffield
Beech Hill Road
Sheffield
S10 2RX
UK
Website: http://mellanbycentre.dept.shef.ac.uk
Tel: 0044114-2713337 (office)
0044114-2713174 (lab)
E-Mail: o.m.gallag...@sheffield.ac.uk
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