Posting for a friend..........
"We have hit a bump in the road with our IHC test and thought maybe you might have some input as we seem to be stuck. We stain (typically for virus) frozen bovine tissues using Biocare's Mach-3 AP polymer kit and develop with Vector Red, counterstain, and dehydrate and mount with Histoclear/VectaMount by hand or xylene/Surgipath mountant on the autocoverslipper. We have been noticing a major loss of stain after development recently. The controls seem to remain stained, it's the more diffuse and minimally stained sections that we have seen stain disappear between development and coverslipping. I did a test with Impress-DAB and Impress-Bajoran Purple to compare and we thought we had solved the problem as being related to the surgipath mountant. However I just did a run with cell markers coverslipping with the Histoclear/Vectamount and there seems to be a loss of stain again. Vector suggested trying 200 mM TBS for wash/diluent/stop instead of 100 mM. This is a recent problem as we have been using this protocol successfully for a few years now. Any suggestions would be greatly appreciated!" Replies directly to meghan.tuc...@ars.usda.gov would be appreciated _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet