[Histonet] Stains for Hemaglobin

[Monfils, Paul]

Years ago I had to do a hemoglobin stain, to demonstrate that round
inclusions in some large cells were in fact phagocytized erythrocytes
(thereby proving that the cells in question were parasitic amebae).  I used
one of two techniques I found in an old book (Histopathologic Technic and
Practical Histochemistry by Lillie and Fullmer, 1976), and whichever
technique I used was quite simple and worked very well. I'm not sure which
one I used (I think it was Dunn-Thompson) so I'll give you both ...


DUNN-THOMPSON METHOD

1. Stain 15 minutes in Mallory's aqueous alum hematoxylin (or other
unacidified alum hematoxylin)
2. Wash in tap water
3. Mordant 1 minute in 4% iron alum
4. Rinse in tap water
5. Stain 15 minutes in picrofuchsin solution (13 ml 1% acid fushsin in 87 ml
saturated aqueous picric acid)
6. Differentiate 3 minutes in 95% ethanol
7. Dehydrate, clear, mount

Results: Hemoglobin emerald green; cytoplasm brown to yellow; collagen red;
nuclei purple to gray-black


OKAJIMA METHOD

1. Mordant 1 minute in 10% phosphomolybdic acid
2. Wash in distilled water
3. Stain 1 hour in ... 9 ml 10% phosphomolybdic acid in 30 ml saturated
aqueous alizarin red
4. Wash in distilled water
5. Counterstain in an unacidified alum hematoxylin , 3 to 5 minutes
6. Wash in water, dehydrate, clear, mount

Results: Hemoglobin orange-red; background light brown