Jennifer, I don't think, that you can substitute Biebrich Scarlett with Eosin. These dyes have a different chemistry. That is well described on Bryan Llewellyn's website: http://stainsfile.info/StainsFile/stain/trichindex.htm
The staining of collagen is also a matter of differentiation of the dyes in water and diluted ethanol. Biebrich Scarlett "leaves" the tissue faster than anilinblue in diluted ethanol. So if you want blue collagen, wash longer in 96%. This will also lead to paler cytoplasm staining. There is a publication on the different dye uptake of collagen, that was exposed to the atmosphere (Histologic 1979). Older blocks show uneven dye uptake. And another publication refers to the tension-state of collagen: "The Masson staining of collagen - an explanation of an apparent paradox" The Histochemical Journal 1975. It says that in tensioned collagen more positivly-charged amino dye-binding sites are available than in relaxed collagen. That leads to better dye uptake of arylmethan dyes of collagen in streched tendon or stretched dermis. I think ripened collagen with many crosslinks has also less dye-binding sites than "younger" collagen. Are there more recent publication on this issue? Gudrun Lang Histolab, Akh Linz -----Ursprüngliche Nachricht----- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Jennifer Anderson Gesendet: Freitag, 23. Oktober 2009 19:52 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Masson's tri-chrome specificity Good morning Histonet. Thanks a lot to so many people who helped me with my Leica XL question - your wealth of knowledge never ceases to impress me. I have a different question now about Masson's tri-chrome and it's specificity on isolated rabbit tendon. There is some muscle still attached to the ends of the tendon. We get both Beibrich Scarlet and Aniline Blue staining, at different intensities, within each collagen sample (even control). How can I explain this? Would there be a possibility to omit the Beibrich and stain with eosin and Aniline blue? Thanks so much! Jennifer M. Anderson, Scientist Halozyme Therapeutics, Inc. 11388 Sorrento Valley Road San Diego, CA 92121 858-704-8333 jander...@halozyme.com<mailto:jander...@halozyme.com> ________________________________ The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. If you received this in error, please contact the sender and delete/destroy this email. _______________________________________________ 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