Lillie's Iron Hematoxylin

Solution A 1940 1954 Function
Hematoxylin 1 g 0.5 g Dye
Ferric chloride 1.2 g 1.2 g Mordant
Ferrous sulphate 4.44 g Mordant
100% ethanol 100 mL Solvent
Distilled water 100 mL 292 mL Solvent
Hydrochloric acid 1 mL 8 mL Solvent


Compounding procedure

Dissolve the ferric chloride in half of the water.
Dissolve the hematoxylin in the other half.
Combine and add the hydrochloric acid.

Dissolve the hematoxylin in the ethanol.
Dissolve the ferric chloride and ferrous sulphate in the water.
Add the hydrochloric acid.
Combine the solutions.


  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for 5-30 minutes.
  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


  1. The 1940 formula is likely not stable for long. If staining is too dark, dip briefly in 1% hydrochloric acid in 70% ethanol.
  2. The 1954 formula is a modification of Wiegert's iron hematoxylin. Lillie states that it is stable for several weeks with occasional use. It is progressive, and does not require differentiation.


Lillie, R.D., (1954)
Histopathologic technique and practical histochemistry Ed.2
Blakiston, New York, USA.

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
    Lillie, (1940)
    Archiv für pathologische Anatomie, v. 29, p. 705.



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