[Histonet] RE: Trichrome following Histofix?

From:"Nicola Cragg"

Hi Paul & Other Histonetters, 

I had the same problem with Trichrome stain on formalin fixed tissue but I found that post-fixing in Bouin's (recommended fix for Trichrome) produces beautiful staining on FFPE sections.  This also worked for Van Gieson's stain, which similarly stains collagen & other connective tissues.  I've just cut & paste the post-fixing part of my protocol below  (I have just used the overnight at room temperature method as I was a bit scared of heating Bouin's up since it contains picric acid) if you want the full protocol let me know: -

1.	Dewax in xylene
2.	Rehydrate through graded alcohols to water
3.	Incubate in Bouin's solution overnight at room temperature or for 1 hour at 56-60*C (sealed container in water bath, open in hood after cooling).
............proceed with stain

Hope this helps!!

Nicola Cragg
EpiStem Ltd
Manchester, UK

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