Hematoxylin Stain–Harris

Reagent Preparation

Mix one part Scott’s Tap Water Substitute with 9 parts deionized water (i.e. one bottle with 900 mL deionized water).


  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Stain in Harris’ Hematoxylin solution for 2 to 2.5 minutes.
  3. Rinse in running tap water.
  4. Dip slides in Differentiating solution 1 to 2 times.
  5. Rinse in running tap water.
  6. Blue in working Scott’s Tap Water Substitute for 1 minute.
  7. Rinse in running tap water.
  8. Stain slides in Eosin Y Alcoholic for 30 seconds.
  9. Dehydrate in alcohol, clear in xylene and mount.


Nuclei should be blue. Nucleoli should be conspicuous and crisply outlined. Cytoplasm will display various shades of pink and red blood cells will be red.


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