[Histonet] Trichrome troubleshooting

Gayle Callis gayle.callis at bresnan.net
Tue Jun 30 15:33:09 CDT 2015


I don't understand what you wrote here " We are using Leica's kit with the
Weigerts iron with Gills".  In a long histo career, I never used Gills half
oxidized hematoxylin for a trichrome stain nor heard of the combined
staining you mentioned.       Could you explain what you are doing here?  Is
this part of the Leica staining protocol?    This is a new one for me to
combine Gills hematoxylin  with Weigerts iron hematoxylin and why it would
even be necessary???   Both of Leica's trichromes kits use Weigerts Iron
hematoxylin and do NOT have Gills in the method.  Their kits look more like
classic Mass Tri methods.   Personally I would only use Weigerts Iron
hematoxylin in a classic Trichrome method.       It is important to know
that iron hematoxylin is not stable over a long period of time since the
ferric chloride continues to oxidize the hematoxylin so it becomes very weak
or doesn't stain.   We never used iron hematoxylin for more than one day
although some people may have success over a week.   We never took a chance
in having weak staining with that continued oxidation by ferric chloride,
and only used freshly mixed iron hematoxylin for a day.   

I would be happy to personally send you a modified Weigerts Iron hematoxylin
that does NOT differentiate out so the iron hematoxylin staining is always
excellent.   It was found in a J of Histotechnology publication many years
ago, and is made up in house - even using other trichrome reagents from the
above vendors I mentioned.   The modified Weigerts iron hematoxylin is more
concentrated compared to the original iron hematoxylin recipe.   

Gayle M. Callis 

-----Original Message-----
From: Suzanne Martin [mailto:smartin at lcpath.com] 
Sent: Tuesday, June 30, 2015 12:37 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Trichrome troubleshooting

Hi all,

We are having trouble troubleshooting our trichrome. It is too blue. We are
using Leica's kit with the Weigerts iron with Gills. Most of the small bowel
controls have seen improvement but patient tissue is not... strange. 

We have tried lessening the time in Gills, adding time for the last acid
step, even lessening time and adding time in the Weigerts. 


Thank you.

Suzanne HT

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