AW: [Histonet] Bone samples long-term storage in 10% formalin or
4% paraformaldehyde
Barry Rittman
barryrittman <@t> gmail.com
Mon Dec 9 07:49:57 CST 2013
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.gallagher <@t> 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.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
> >
> >
> > *STOP*: Do you really need to print this e-mail?
> >
> > *BE GREEN:* Keep it on the screen.
> >
> >
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> >
> >
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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
> you have received.
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