Heidenhain's Iron Hematoxylin

Solution A Amount Function
Ferric ammonium sulphate 2.5 g Mordant
Distilled water 100 mL Solvent
Solution B Amount Function
Hematoxylin 0.5 g Dye
Distilled water 100 mL Solvent


Compounding procedure
Make each solution seperately.
Solution B should be ripened for a minimum of one month.


  1. Bring sections to water with xylene and ethanol.
  2. Place into solution A for 30 minutes to 24 hours.
  3. Rinse with distilled water.
  4. Place into solution B for 30 minutes to 24 hours.
  5. Rinse with tap water.
  6. Differentiate in solution A, controlling microscopically.
  7. Wash well in running tap water to blue.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results


  1. The stock solutions are stable for some time.
  2. The hematoxylin solution needs to be ripened.
  3. Staining at elevated temperatures (not over 60°C) will shorten the required times. Always differentiate at room temperature.
  4. The degree of differentiation will determine which tissue components are prominent. The method can demonstrate many structures, including chromosomes, nuclear components, mitochondria and muscle striations
  5. The solutions may be reused, with the exception of the solution A used to differentiate, which should be fresh each time.
  6. Counterstaining is not recommended.
  7. This method is usually recommended for monochrome photography.


Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
    Heidenhain, M., (1892).
    Festschrift Herrn A. von Kolloker zur Feier seines fünfzigjährigen medicinischen
    Wilhelm Engellmans, Leipzig, Germany



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