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Krutsay’s Alum & Iron Hematoxylin

Krutsay's Alum & Iron Hematoxylin


Krutsay’s Alum hematoxylin gives very highly selective staining of nuclei. It may also be easily converted into an iron hematoxylin for use as an acid resistant nuclear stain.


Alum hematoxylin

Hematoxylin1 gDye
Potassium alum50 gMordant
Distilled water1 LSolvent
Hydrochloric acid5 mLAcidifier
Sodium iodate0.2 gOxidant

Iron conversion

Iron alum2 gMordant
Hydrochloric acid0.5 mLAcidifer
Distilled water100 mLSolvent

Compounding Procedure

  1. Mix the reagents under Alum hematoxylin together.
  2. Bring to a boil
  3. Cool and filter.
  4. The solution may be used immediately.

Conversion to Iron Hematoxylin

  1. Mix the reagents under Iron conversion together, and dissolve.
  2. For use mix the following:
    • Alum hematoxylin solution – 25 mL
    • Iron conversion solution – 2 mL


  1. Bring sections to water with xylene and ethanol.
  2. Place into either staining solution for 5 minutes.
  3. Rinse with water and blue.
  4. Rinse well with water.
  5. Counterstain if desired.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue (alum) or black (iron)
  • Background  –  as counterstain or unstained


  • This is a progressive solution and is highly selective for nuclei.
  • The iron Alum is (FeNH4(SO4)2.12 H2O)
  • 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. Humason, G. L.
    Animal Tissue Techniques.
    W. H. Freeman and Co., San Francisco, CA, USA