[Histonet] Masson's tri-chrome specificity

[Lee & Peggy Wenk]

Try extending the time in Bouins. Tendon is rather dense, and may need
longer time in the post-mordant. 60 degrees C for 1 hour, with a good 5-10
minute wash in running water afterwards, to remove all the yellow.

The same with the phosphotungstic acid rinse between the red and blue dyes.
Make it 5 minutes.

Other substitutes for Biebrich scarlet are Ponceau de xylidene, Ponceau S,
Acid fuchsin, Chromotrope 2R. 

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Saturday, October 24, 2009 4:45 AM
To: 'Jennifer Anderson'
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: AW: [Histonet] Masson's tri-chrome specificity

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-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Jennifer
Anderson
Gesendet: Freitag, 23. Oktober 2009 19:52
An: histonet <@t> 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
janderson <@t> halozyme.com<mailto:janderson <@t> halozyme.com>



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