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Mayer’s Alum Hematoxylin Staining Variants

Mayer's Alum Hematoxylin Staining Variants


The solution usually meant when Mayer’s hemalum is specified is actually a modification of Mayer’s 1901 formula by Langeron. Mayer published several alum hematoxylin variants for nuclear staining, both progressive and regressive.

Such solutions usually contain hematoxylin and an alum, and are called hemalum or alum hematoxylin solutions. Many formulae have been suggested. They vary in the amount of hematoxylin, the amount and type of aluminum salts, solvents, oxidizing agents and stabilizers.


This protocol follows the standard hematoxylin staining procedure with any of the five alum hematoxylin variants substituted in the second step. The choice of variant depends on the desired outcome.

Hematoxylin1 g1 g4 g1 g1 gDye
Ammonium alum50 g50 g50 g50 gMordant
Potassium alum50 gMordant
Distilled water1 L1 L700 mL1 L1 LSolvent
95% ethanol50 mL50 mLSolvent
Glycerol300 mLStabilizer
Sodium iodate0.2 g0.2 gOxidant
Glacial acetic acid20 mLAcidifier

Compounding procedures

To prepare the variant that you will be using for your hematoxylin staining, follow these compounding procedures.


  1. Dissolve the hematoxylin in ethanol.
  2. Dissolve the Alum in water.
  3. Combine.
  4. Ripen for months.
  5. Add the acetic acid to 1891b


  1. Grind the hematoxylin with some glycerol.
  2. Mix all the other constituents together.
  3. Add the hematoxylin paste.
  4. Wash out the paste with the solution.
  5. Ripen for months.
  6. This formula produces a regressive stain.

1901, 1903

  1. Dissolve the Alum and hematoxylin in the water.
  2. Add the sodium iodate.
  3. Bring to a boil. Cool.


  1. Bring the sections to water
  2. Place in a variant for a few minutes (start with 5 minutes)
    1. If using the 1896 variant, rinse with tap water, then
    2. Differentiate with acid alcohol.
  3. Wash well with tap water to blue.
  4. Counterstain with eosin.
  5. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei – blue
  • Mucins – unstained (blue if using mucihematein)
  • Background – unstained


  • The 1891, 1901 and 1903 formulae are progressive.
  • The 1896 formula is regressive.
  • The 1891 and 1896 formulae are to be ripened naturally.
  • The 1901 and 1903 formulae can be used immediately.
  • The 1896 formula is also known as Mayer’s glychemalum.

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.