This is a cached page for the URL ( To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
Protocol: Hematoxylin Stain. [OHSU Intranet] [Search the OHSU Site] [OHSU Site Map] [Oregon Health Sciences University] [Neurological Sciences Institute] [Faculty] [Education] [Public] [Seminars] [Contact] [Directions] OHSU Website (NavBar): the Neurological Sciences Institute


Hematoxylin Stain



Hematoxylin 6.4g
Ammonium alum 60g
Ethanol 200ml
Glycerol 160ml
Distilled water 640ml

Mix for a day then allow the solution to ripen in the dark for at least a month before using.

Acidic Alcohol

Concentrated HCl 4ml
95% Ethanol 396ml

Bluing Agent

Sodium bicarbonate 1g
Distilled water 1 l


  1. Deparaffin and rehydrate slides.
  2. Slightly overstain the sections with hematoxylin 3-5 minutes.
  3. Remove excess stain in tap water for 2 minutes.
  4. Differentiate and restain a few seconds in acidic alcohol until sections look red.
  5. Rinse in tap water.
  6. Blue in bicarbonate until nuclei stand out sharply blue for 2 minutes.
  7. Rinse in running tap water.
  8. Dehydrate and clear, coverslip with Cytoseal.

Valid HTML!


This document was created for the neurogenetics laboratory run by P. H. Reddy, Ph.D. in the NSI of OHSU. Page last updated: 18 Sept 2001.