AW: [Histonet] Bone samples long-term storage in 10% formalin or
4% paraformaldehyde
Orla M Gallagher
o.m.gallagher <@t> sheffield.ac.uk
Mon Dec 9 07:44:47 CST 2013
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.Wineman <@t> 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.wineman <@t> novusint.com
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of pruegg <@t> ihctech.net
> Sent: Thursday, December 05, 2013 2:50 PM
> To: gu.lang <@t> gmx.at; 'Orla M Gallagher'
> Cc: histonet <@t> 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.lang <@t> gmx.at>
> Date: 12/5/13 11:42 am
> To: "'Orla M Gallagher'" <o.m.gallagher <@t> sheffield.ac.uk>
> Cc: histonet <@t> 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
>
> -----Ursprüngliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Orla M
> Gallagher
> Gesendet: Donnerstag, 05. Dezember 2013 19:31
> An: histonet <@t> 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.gallagher <@t> sheffield.ac.uk
>
>
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--
**************************
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.gallagher <@t> sheffield.ac.uk
*STOP*: Do you really need to print this e-mail?
*BE GREEN:* Keep it on the screen.
*Times Higher Education University of the Year*
Data protection and confidentiality:
The information contained in this message or any appended documents may be
privileged and confidential and is intended for the exclusive use of the
addressee(s). If you are not the addressee, any disclosure, reproduction,
distributions, other dissemination or use of this message is strictly
prohibited and may be unlawful. If you receive this correspondence in error
please contact the sender immediately and permanently delete/destroy what
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