Frozen section
Staining slides


Topic Completed: 1 October 2011

Revised: 03 January 2019

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PubMed Search: Staining slides


Jessica Wallace, P.A., A.S.C.P.
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Cite this page: Wallace J. Staining slides. PathologyOutlines.com website. http://www.pathologyoutlines.com/topic/frozensectionslideorder.html. Accessed June 24th, 2019.
Procedure for staining slides
  • Keep all stains and solutions fresh and well maintained
  • Dip slide in reagents in this order for H&E staining:
    • After obtaining frozen section, IMMEDIATELY fix in 95% ethanol (even 15 seconds of delay can cause significant artifact)
    • Formal alcohol, formalin or 95% alcohol: 45 - 60 seconds
    • Water: 5 - 7 seconds
    • Hematoxylin: 60 seconds
    • Lithium carbonate or 0.2 % aqueous ammonia (Bluing): 15 - 20 seconds
    • Eosin: 20 - 60 seconds
    • 95% alcohol: 10 seconds
    • 100% alcohol: 10 seconds
    • Xylene, toluene, limonene derivatives and Clearite: 10 seconds
    • Then add mounting media for cover slipping
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