Hello Histonetters, We regularly perform the Masson Trichrome stain in our lab and recently have been noticing some inconsistent results. The main issue seems to be that the Biebrich Scarlet is not being adequately removed by the PTA/PMA, as many of the areas containing collagen are showing up as purple instead of bright blue. We use fresh reagents every time we run the stain and our protocols are as follows:
Rehydrate (xylene --> 100%EtOH --> 95% --> 80% --> 70%) Bouin's in microwave (1 min) Stand at RT in Bouin's (15 min) Running tap (5 min) Weigert's Hx (10 min) Running Tap (5 min) Biebrich Scarlet (1.25 min) Rinse dH2O Phosphotungstic acid/Phosphomolybdic acid (10 min) Aniline Blue (10 min) Rinse dH2O 1% Acetic acid (1 min) Rinse dH2O Air dry for dehydration Has anyone else experienced this same problem and, if so, what corrective actions have worked for you? I'd greatly appreciate your advice! Thanks, Anna -- Anna Coffey Senior Histology Technician Department of Oncology Histopathology and Tissue Shared Resource LR-10 Pre-Clinical Sciences Building Lombardi Comprehensive Cancer Center Georgetown University 202-687-7890 ah...@georgetown.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet