[Histonet] Masson Trichrome, Alligator Heart Tissue, colors turning out purple

cscampbe <@t> uci.edu cscampbe <@t> uci.edu
Tue Mar 9 19:06:21 CST 2010

To Histonet:

This is my first time mailing the histonet community. Coincidentally, this
is my first venture into histology. My lab is conducting research on
alligator heart tissue. At the moment, I am attempting to stain my slides
using Masson Trichrome to differentiate muscle from connective tissue.

Results have not been optimal. The slides turn out to be more purple than
I had anticipated (comparing to pictures from journals). The red can be
told apart from the teal, but the colors are not bright.

Here is the procedure I am using:

1. Deparaffinize and hydrate to water
2. Iron Alum, 30 min
3. Wash in Dis. Water, 5 min
4. Hematoxylin stain, 30 min
5. Wash in Dis. Water, 5 min
6. Differentiate in saturated aq. picric acid, 1 min
7. Wash in dis. water, 10 min
8. Stain in acid fuschin, 5 min
9. Rinse in water
10. Stain in Ponceau de Xylidene, 5 min
11. Rinse in water
12. Differentiate in Phosphomolybdic acid, 5 min
13. Fast Green, 2 min
14. Differentiate in 1% aq. acetic acid, 5 min
15. Dehydrate
16. xylene (two washes, 3 min each)

I would greatly appreciate any advice on how to modify my procedure to
better stain the cardiac tissue. Or should I be using a different
trichrome method better suited to this particular tissue?

Thank you!

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