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Anderson’s Iron Hematoxylin

Anderson's Iron Hematoxylin

6
steps
7
materials

Materials

Solution A

MaterialAmountFunction
Hematoxylin0.5 gDye
100% ethanol50 mLSolvent
Distilled water50 mLSolvent
Calcium hypochlorite, 2% aqueous5 mLOxidant

Solution B

MaterialAmountFunction
Ferric ammonium sulphate3 gMordant
Distilled water100 mLSolvent
Sulfuric acid0.5 mLAcidifier

Compounding procedures

  1. Make each solution separately.
  2. For use, add 2 volumes of solution A to 1 volume of solution B.
  3. The working solution may be used immediately, but is not stable for long.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for an appropriate time.
  3. Wash well in running tap water to blue.
  4. Rinse with distilled water.
  5. Counterstain if desired.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Background  –  as counterstain or unstained

Notes

  • The stock solutions are stable for some time.
  • The working solution should be made fresh.
  • It is claimed that the working solution rarely overstains, i.e. it is progressive.
  • Solution A incorporates calcium hypochlorite as an oxidizing agent for hematoxylin. Presumably other oxidizing agents would suffice equally as well. Sodium iodate (0.1 g or less) is the most common.
  • The staining time should be determined by trial.

Safety Note

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

References

  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Citing:
    Anderson, J., (1929)
    How to stain the nervous system.
    Livingstone. Edinburgh, Scotland.