[Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde

Sean McBride smcbride <@t> andrew.cmu.edu
Mon Dec 9 10:46:24 CST 2013 

Dear Orla,

Post fixation, we have stored our bone specimens in 1x PBS while having them sent out for MicroCT analysis.  We have also stored them in 1x PBS at 4°C post fixation when necessary until further processing without having adverse affects on our staining.  However, we chose conventional staining over IHC for our results.


Best regards,


~Sean McBride


Scientific Specialist
Bone Tissue Engineering Center
Carnegie Mellon Research Institute
Suite 4311
700 Technology Drive
Pittsburgh, PA 15219-3124

412-268-8275 (o)
412-915-1683 (m)
412-268-8275 (fax)
smcbride <@t> andrew.cmu.edu 






On Dec 9, 2013, at 8:44 AM, Orla M Gallagher 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
>> 
>> -----Urspr&uuml;ngliche 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.
>> 
>> 
>> *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.
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> 
>> 
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> 
> 
> 
> 
> -- 
> **************************
> 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.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

More information about the Histonet mailing list