Hematoxylin Stain–Gill

Reagent Preparation

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

Formalin-Ethanol Fixative (Films)

Mix 5 mL of formaldehyde with 45 ml of 95% ethanol. Prepare fresh.


  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Stain in Gill's Hematoxylin No. 2 or No. 3 solution for 1.5 to 3 minutes.
  3. Rinse in running tap water.
  4. Blue in working Scott’s Tap Water Substitute for 1 minute.
  5. Rinse in running tap water.
  6. Stain slides in Eosin Y Alcoholic for 30 seconds.
  7. Place slides directly in 95% alcohol for 15 seconds.
  8. Dehydrate in alcohol, clear in xylene and mount.


Chromatin appears blue to blue-black and nucleoli should be conspicuous. Cytoplasmic staining with Gill hematoxylin solutions should be pale or absent, thus, acid differentiation may not be necessary. Cytoplasm should stain with different shades of pink with eosin.

Hematoxylin Stain-Gill


Related Links