Lillie’s modification of Weigert’s iron hematoxylin

MATERIALS

Ethanol (95%)

Ferric chloride (FeCl₃•6H₂O)

Ferrous sulfate (FeSO₄•7H₂O)

Hematoxylin

Hydrochloric acid (concentrated)

METHOD

• 1. Make 1% alcoholic hematoxylin by dissolving 1.0 g of hematoxylin in 100 mL of 95% ethanol.

• 2. Combine the following reagents and stir until dissolved (filter if necessary, to remove undissolved impurities):

  • 2.5 g ferric chloride

  • 4.5 g ferrous sulfate

  • 298 mL H₂O

  • 2.0 mL concentrated hydrochloric acid

• 3. Add the solution containing the iron salts to the alcoholic hematoxylin solution.

  The mixture goes black. Lillie’s solution is stable for 2-3 mo at room temperature.