Iron pigments - bright blue
Nuclei – dark pink to red
Cytoplasm - light pink
Figure 1. Positive control dog splenic hemosiderosis.
Mallory’s Iron Staining Procedure
Purpose:
This standard operating procedure (SOP) establishes the standard for staining
iron pigment using the Mallory’s Iron staining procedure.
Reagents and Materials:
Reagents
5% Potassium Ferrocyanide
Potassium Ferrocyanide 25gm
Distilled water 500ml
5% Hydrochloric Acid
Hydrochloric Acid 25ml
Distilled water 475ml
Make fresh:
5% Hydrochloric Acid 20ml
5% Potassium Ferrocyanide 20ml
Nuclear Fast Red
Commercially prepared by Newcomer Supply Co.
(catalog # 12552)
Expiration date provided by manufacturer.
Store at room temperature.
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and synthetic mounting media.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample
Preparations:
Paraffin tissue section slides cut at 4 microns.
Calibration
and Standardization:
A tissue section slide containing iron pigment is stained with each procedure.
Procedure:
1. Deparaffinize
and hydrate to distilled water.
2. Iron Stain - 10 minutes.
Make fresh:
5% Hydrochloric Acid 20ml
5% Potassium Ferrocyanide 20ml
3. Distilled water rinse.
4. Nuclear Fast Red - 5 minutes.
5. Distilled water rinse.
6. Dehydrate, clear and coverslip.
Make fresh:
Light green 10ml
Distilled water 40ml
11. Distilled water rinse.
12. Dehydrate, clear and coverslip.
Positive
control: dog spleen with hemosiderosis
Results:
Iron pigments - bright blue
Nuclei – dark pink to red
Cytoplasm - light pink
Figure 1. Positive control dog splenic hemosiderosis.
Effective:
July 1, 2004
Cherie Chapman
James R. Turk
References:
Sheehan DC, Hrapchak BR: Theory and Practice of Histotechnology, 2nd Ed, CV Mosby, St. Louis, 1980, pp. 217-218.Armed Forces Institute of Pathology Laboratory Methods in Histotechnology, eds. Prophet EB, Mills B, Arrington JB, Sobin LH, American Registry of pathology, Washington D.C. , 1992, pp. 196.