AW: [Histonet] Bone/cartilage/epithelial tissue stain

[Gudrun Lang]

Rui,
in your first post about your problem, you wrote about decalcification of
your samples. 
Are these samples, where you want to demonstrate bone, decalcified? Which
decalcifier?
Haematoxylin stains only non-decalcified bone, due to the Ca-ions.
Strong acids for decalcification (like HNO3 or HCl) may alter the
tissue-stainability. They are said to solve some amount of proteoglycans.
If they are not decalcified, you can combine a von-Kossa-stain with a
trichrome-stain or alcianblue. Calcified bone stains black.

An easy stain would be the combined Alcianblue-PAS. Alcianblue demonstrates
acid proteoglycans, PAS demonstrates neutral glycoproteins like collagen.
But I think, proteoglycans and collagen are in such a tight junction in
cardilage, that it will give a mixed dye-appearance. But worth a try.

Gudrun

-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Rui TAHARA
Gesendet: Freitag, 23. August 2013 09:49
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Bone/cartilage/epithelial tissue stain



Hi,
Would anyone suggest me what staining is best to color differentiate between
cartilage and bone and epithelial tissues in avian embryos? 

I have been trying Mallory Trichrome for embryos but recently I was
suggested that Mallory Trichrome stains cartilage differently in embryos
compared to adult samples since Aniline blue stains fiber that may not
develop in early embryos. There is some protocol that modified the Mallory
Trichrome that could be applied to embryos. However, the resulting colors of
all tissues look all purple-ish and difficult to tell the cartilage from the
weak blue stain from aniline blue. 

Currently I am thinking to try out Alcian blue/Hematoxylin and Eosin stain
(Ehrlich’s hematoxylin). The purpose of the staining is to look at
interaction between ossification and epithelial development so I think
alcian blue for staining cartilage works but I am wondering if there is any
other staining combination with alcian blue exist for visualizing bone and
epithelial tissue (e,g. alcian blue/alizarine red with other staining?). 

 

Any suggestion would be appreciated! 

Rui TAHARA
PhD Candidate
Biology Department
McGill University