I have been staining mouse femurs that were decalcified in 0.5M EDTA at 4C for 3 weeks using a TRAP protocol from University of Rochester Medical Center TRAP staining paraffin sections. I see very nice bright red osteoclasts after 30 minutes in the staining solution. However sometime between this step and mounting, the red stain is totally removed. I initially rehyrdated in dips of 70, 90, 100, 100 EtOH, then 3x 3min in xylene. I then tried quick dips in each, including xylene, and it seems to be a little better. Does xylene remove the fast red violet stain?