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 > > > > -----Ursprüngliche 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 > > > > > > *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@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > > Histonet mailing list > > Histonet@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.gallag...@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). 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