PAP labeling enhancement by osmium tetroxide-potassium ferrocyanide treatment

J Histochem Cytochem. 1988 Jun;36(6):697-9. doi: 10.1177/36.6.3284949.

Abstract

We describe a procedure to intensify staining of antigens labeled by the peroxidase-antiperoxidase (PAP) method. Routinely PAP-stained samples were enhanced by application of 5% osmium tetroxide for 30 min followed by freshly prepared 2% potassium ferrocyanide, plus 1% hydrochloric acid for 15 min. The staining color changed from the original golden-brown, through transient gray, to a very dark brown that was almost black. In addition to stronger labeling, antigen locations not apparent in untreated specimens may thus be disclosed. Furthermore, retrospective staining intensification can be performed in stored PAP-labeled samples even years later.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation, T-Lymphocyte / analysis
  • Antigens, Viral / analysis
  • Astrocytes / metabolism
  • Ferrocyanides*
  • Immunoenzyme Techniques*
  • Immunohistochemistry / methods*
  • Osmium Tetroxide*
  • Osmium*
  • Rats
  • Spleen / cytology
  • Vimentin / metabolism

Substances

  • Antigens, Differentiation, T-Lymphocyte
  • Antigens, Viral
  • Ferrocyanides
  • Vimentin
  • Osmium
  • potassium ferrocyanide
  • Osmium Tetroxide