AW: [Histonet] holly nuclei bat man
I have a quiet semi-scientific explanation. Formalin-fixation stabilizes proteins via crosslinking. AR solves methylol-adducts due to formaldehyde and perhaps also some crosslinks. Nuclei also have a protein-scaffold, filled with nucleinacids. It seems like a ladder or hole in tights. It seems like there is a kind of tension, after breaking a crosslink the hole blubbs out. This is also seen with pretreatment in hybridization-procedures. The more enzymatic and heat-retrieval the more holes. And there is a direct relationsship between fixation-quality and AR-strength. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Patsy Ruegg Gesendet: Samstag, 05. Jänner 2013 21:38 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] holly nuclei bat man Happy New Year everyone! Can we start a discussion once again on what causes holes in nuclei? It is pretty clear in my experience it has something to do with heating at least for me with antigen retrieval. When I do HIER at ph 8 especially I am seeing a lot of holes in nuclei. HE on same tissue alone without HIER/IHC doesn't seem to exhibit this artifact (holes in the nuclei). I bet it can also happen if the tissue is not fixed well enough to protect it from paraffin processing but right now I am seeing it more after HIER, don't see it as much on my IHC slides with no AR or when I use eier instead of hier. Also notice that ph6 HIER and Ph9 HIER are not as bad as when I use Ph8 for the same times and temps. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 mailto:pru...@ihctech.net pru...@ihctech.net ___ 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] a problem with giemsa stain for h.pylori
hello every body, on giema staining of gastric tissue slides for h.pylori,we have encounterd a major problem for a while.the problem is deposition of large flakes on the mucosal surface and we do not know why? do you know how to get rid of this problem. thank you in advance for your kind help. kind regards, shahram. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] holly nuclei bat man
According with some tests I made and an article published in the JOH, the so called nuclear holes occur when a section with some remaining amount of water underneath is dried in an oven at 60ºC or more. The explanation is that the heated underlying water dissolves nuclear material and hence the appearance of holes. I could look for the reference of the JOH article if you want. René J. From: Patsy Ruegg pru...@ihctech.net To: histonet@lists.utsouthwestern.edu Sent: Saturday, January 5, 2013 3:37 PM Subject: [Histonet] holly nuclei bat man Happy New Year everyone! Can we start a discussion once again on what causes holes in nuclei? It is pretty clear in my experience it has something to do with heating at least for me with antigen retrieval. When I do HIER at ph 8 especially I am seeing a lot of holes in nuclei. HE on same tissue alone without HIER/IHC doesn't seem to exhibit this artifact (holes in the nuclei). I bet it can also happen if the tissue is not fixed well enough to protect it from paraffin processing but right now I am seeing it more after HIER, don't see it as much on my IHC slides with no AR or when I use eier instead of hier. Also notice that ph6 HIER and Ph9 HIER are not as bad as when I use Ph8 for the same times and temps. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 mailto:pru...@ihctech.net pru...@ihctech.net ___ 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: [Histonet] a problem with giemsa stain for h.pylori
Filter your Giemsa solution before using it or better yet, get a newer staining solution René J. From: Shahram Sabeti sabeti_shah...@yahoo.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Sunday, January 6, 2013 10:07 AM Subject: [Histonet] a problem with giemsa stain for h.pylori hello every body, on giema staining of gastric tissue slides for h.pylori,we have encounterd a major problem for a while.the problem is deposition of large flakes on the mucosal surface and we do not know why? do you know how to get rid of this problem. thank you in advance for your kind help. kind regards, shahram. ___ 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: [Histonet] holly nuclei bat man
Yes this all makes good sense and explains why I may not be able to have control of this because I do not always do the tissue fixation and processing myself. That first step of adequately fixing (cross linking the proteins) to protect them from tissue processing and AR are so important. The one thing I can control is to make sure the slides are completely dry before baking them as Renee mentioned. Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 pru...@ihctech.net -Original Message- From: Gudrun Lang [mailto:gu.l...@gmx.at] Sent: Sunday, January 06, 2013 4:32 AM To: 'Patsy Ruegg' Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] holly nuclei bat man I have a quiet semi-scientific explanation. Formalin-fixation stabilizes proteins via crosslinking. AR solves methylol-adducts due to formaldehyde and perhaps also some crosslinks. Nuclei also have a protein-scaffold, filled with nucleinacids. It seems like a ladder or hole in tights. It seems like there is a kind of tension, after breaking a crosslink the hole blubbs out. This is also seen with pretreatment in hybridization-procedures. The more enzymatic and heat-retrieval the more holes. And there is a direct relationsship between fixation-quality and AR-strength. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Patsy Ruegg Gesendet: Samstag, 05. Jänner 2013 21:38 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] holly nuclei bat man Happy New Year everyone! Can we start a discussion once again on what causes holes in nuclei? It is pretty clear in my experience it has something to do with heating at least for me with antigen retrieval. When I do HIER at ph 8 especially I am seeing a lot of holes in nuclei. HE on same tissue alone without HIER/IHC doesn't seem to exhibit this artifact (holes in the nuclei). I bet it can also happen if the tissue is not fixed well enough to protect it from paraffin processing but right now I am seeing it more after HIER, don't see it as much on my IHC slides with no AR or when I use eier instead of hier. Also notice that ph6 HIER and Ph9 HIER are not as bad as when I use Ph8 for the same times and temps. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 mailto:pru...@ihctech.net pru...@ihctech.net ___ 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: [Histonet] holly nuclei bat man
The reference for the Nuclear bubbling article is: Journal of Histotechnology, 1990; 13(2):135-6 René J. From: Patsy Ruegg pru...@ihctech.net To: gu.l...@gmx.at Cc: histonet@lists.utsouthwestern.edu Sent: Sunday, January 6, 2013 12:34 PM Subject: RE: [Histonet] holly nuclei bat man Yes this all makes good sense and explains why I may not be able to have control of this because I do not always do the tissue fixation and processing myself. That first step of adequately fixing (cross linking the proteins) to protect them from tissue processing and AR are so important. The one thing I can control is to make sure the slides are completely dry before baking them as Renee mentioned. Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 pru...@ihctech.net -Original Message- From: Gudrun Lang [mailto:gu.l...@gmx.at] Sent: Sunday, January 06, 2013 4:32 AM To: 'Patsy Ruegg' Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] holly nuclei bat man I have a quiet semi-scientific explanation. Formalin-fixation stabilizes proteins via crosslinking. AR solves methylol-adducts due to formaldehyde and perhaps also some crosslinks. Nuclei also have a protein-scaffold, filled with nucleinacids. It seems like a ladder or hole in tights. It seems like there is a kind of tension, after breaking a crosslink the hole blubbs out. This is also seen with pretreatment in hybridization-procedures. The more enzymatic and heat-retrieval the more holes. And there is a direct relationsship between fixation-quality and AR-strength. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Patsy Ruegg Gesendet: Samstag, 05. Jänner 2013 21:38 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] holly nuclei bat man Happy New Year everyone! Can we start a discussion once again on what causes holes in nuclei? It is pretty clear in my experience it has something to do with heating at least for me with antigen retrieval. When I do HIER at ph 8 especially I am seeing a lot of holes in nuclei. HE on same tissue alone without HIER/IHC doesn't seem to exhibit this artifact (holes in the nuclei). I bet it can also happen if the tissue is not fixed well enough to protect it from paraffin processing but right now I am seeing it more after HIER, don't see it as much on my IHC slides with no AR or when I use eier instead of hier. Also notice that ph6 HIER and Ph9 HIER are not as bad as when I use Ph8 for the same times and temps. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Fax: 720-859-4110 mailto:pru...@ihctech.net pru...@ihctech.net ___ 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] Re: holly nuclei bat man
Hmmm...I have raised this issue beforeno answer. All answers to datepure conjecture! Lol...if you meant Holly.rather than Holy or even Holey...nice one, Batwoman! Caped Crusader. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
