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Sass’ Alum Hematoxylin Variants

Sass' Alum Hematoxylin Variants



MaterialVariation IVariation IIFunction
Hematoxylin10 g1 gDye
Ammonium alumsaturated50 gMordant
Distilled water1 L1 LSolvent
Sodium iodate10 g1 gOxidant
Glacial acetic acid30 mLAcidifier

Compounding Procedure

  1. Dissolve the Alum in the water.
  2. Add the hematoxylin.
  3. Add the other ingredients.
  4. Filter.
  5. The solutions may be used immediately.


  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

  • Nuclei  –  blue
  • Background  –  as counterstain or unstained


  • Both of these formulae use 1 gram of sodium iodate for each gram of hematoxylin. However, 1 gram of hematoxylin is oxidized by 0.2 grams of sodium iodate, so the solutions should be expected to have a relatively short life.
  • Due to it’s high dye content, the Var I formula is likely regressive.
  • The Var II formula is likely progressive.
  • The staining time should be determined by trial.
  • Acid ethanol is 0.5% – 1% hydrochloric acid in 70% ethanol.
  • 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.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.


  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.