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Tri-Crome stain always purple instead of red

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#1 Empty_H



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Posted 22 October 2014 - 12:29 PM

I am doing a Tri-Crome stain on frozen mouse lung samples. The lungs should be red and any fibrosis should be blue (with cells being much smaller and darker). The problem I have is it always turns out purple and blue instead of red and blue. Any suggestions?


Normal Procedure:

ON Bouin's solution @ RT

2min rinse in tap water

5 minutes Working Weigert's Iron Hematoxylin (mixed just beforehand)

10 minutes rinse in tap water

Rinse in DI water

5 minutes Biebrich Scarlet-Acid Fucshin

3x1 rinse in tap water

Rince in DI Water

3x3 minutes Phosphotungstic/Phosphomolybdidc acid solutions

5 minutes Aniline Blue solution

3x1 minute rinse in tap water

2 minutes Acetic Acid 1%

2x1 minute rinse in tap water

rinse in DI water


Multiple 100% iso washes

Multiple Xylene washes


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