Hi Sharon, HPS is our routine stain (instead of H&E, which 98% of labs use). Not sure what part of the stain you are having trouble with, but most unpredictability usually comes in the phloxine/saffron balance. Some things to watch for are: -keeping your saffron covered, and I would suggest parafilm-ing it when not in use -keep the alcohols before the saffron rigorously free of phloxine-contaminated alcohol, thus not allowing tainted alcohol to contaminate/dilute the saffron during staining -getting all the background hematoxylin and phloxine out, which can be checked as you go along when you validate your method -be diligent about the preparation of the saffron. In addition to Hermina's suggested method, we microwave the dry saffron, grind it, then boil the alcoholic mixture to remove as much moisture as possible What do you use the stain for, i.e. differentiating muscle and collagen? As a routine stain, I understand its use in Canada is limited to a few laboratories in the east such as the Ottawa, Kingston, Sudbury areas. This is often because pathologists who trained here at Queen's Pathology like to take the stain with them. Having said that, many of our residents request slides stained with H&E, to familiarize themselves with that stain before exams. Good luck, hope this helps... Eric Gagnon MLT Histology Laboratory, Kingston General Hospital Kingston, Ontario, Canada
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