Suzanne, 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 HTL/HT/MT(ASCP) -----Original Message----- From: Suzanne Martin [mailto:[email protected]] Sent: Tuesday, June 30, 2015 12:37 PM To: [email protected] 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. Thoughts? Thank you. Suzanne HT _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
