Bullard's Alum Hematoxylin

Ingredient Amount Function
Hematoxylin 8 g Dye
Ammonium alum 20 g Mordant
Distilled water 250 mL Solvent
50% ethanol 144 mL Solvent
Glacial acetic acid 16 mL Acidifier
Mercuric oxide, red 8 g Oxidant
95% ethanol 275 mL Solvent
Glycerol 330 mL Stabiliser
Glacial acetic acid 18 mL Acidifier
Ammonium alum 40 g Mordant


Compounding procedure
Combine the first five ingredients and bring to the boil.
Add the mercuric oxide with caution.
Cool and filter.
Add the last four ingredients.
The solution may be used immediately, and is stable for more than a year.


  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for an appropriate time.
  3. Rinse well with water.
  4. Differentiate with acid ethanol if necessary.
  5. Rinse with water and blue.
  6. Rinse well with water.
  7. Counterstain if desired.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results


  1. The solution is likely regressive
  2. li>The appropriate time should be determined by trial.
  3. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
  4. Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.

F. A. Putt
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA




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