Re: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
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 *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
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 *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
RE: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
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 *STOP*: Do you really need to print this e-mail? *BE GREEN:* Keep it on the screen. *Times Higher Education University
Re: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
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) smcbr...@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.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 *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
RE: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
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 *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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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
AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
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
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 *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