Lillie's Method For Ferric and Ferrous Iron

EMS26604

Components:

Potassium Ferrocyanide

Potassium Ferricyanide

Hydrochloric Acid 0.5%

Basic Fuchsin 0.5%

Acetic Acid 1% Aqueous

Fixation:

10% Buffered Neutral Formalin

Sections:

Paraffin @ 6µm

Staining Procedures:

(Use control slide. Use chemically clean glassware)

a) Preparation of Potassium Ferrocyanide Solution:

Prepare fresh, just before use

• Potassium Ferrocyanide - 0.4g
• Hydrochloric Acid - 40.0ml

b) Preparation of Potassium Ferricyanide Solution:

Prepare fresh just before use.

• Potassium Ferricyanide - 0.4g
• Hydrochloric Acid - 40.0ml

1. Deparaffinise and hydrate to distilled water.
2. For Ferric Iron, place in Potassium Ferrocyanide Solution for 1 hour. For Ferrous Iron, place in Potassium Ferricyanide Solution for I hour.
3. Wash in Acetic Acid, 1%.
4. Stain for 10 minutes in Basic Fuchsin Solution. Rinse in distilled water, (For critical work, omit this step).
5. Dehydrate in 95% alcohol, absolute alcohol and clear in xylene, two changes each.
6. Mount with Permount.

Stain Results:

References:

Lillie, R.D., Histopathologic Technic and Practical Histochemistry, 3rd ed. McGraw-Hill, N.Y., p.407, c 1965.

Luna, L.G. (ed). Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology, 3rd ed., McGraw-Hill, N.Y., c. 1968, p.182.