Hirano-Zimmerman for Nerve Cells & Fibres

EMS26698

Components:

Silver Nitrate 10% Aqueous

Ammonia Water 0.1%

Formalin Solution 50%

Gold Chloride 0.05% Aqueous

Sodium Thiosulfate 5% Aqueous

Fixation:

Formalin 10% Neutral Buffered

Sections:

Paraffin @ 12 – 15µm

Staining Procedures:

1. Deparaffinise and hydrate to distilled water.
2. Silver Nitrate, 10% for two hours or longer.
3. Place sections directly in Ammonia Water for 3 minutes.
4. Place section directly in Formalin Solution 3 minutes. Tissue becomes black-brown. Rinse in distilled water, two changes.
5. Place section again into Silver Nitrate, 10% for 3 – 5 minutes.
6. Repeat step 3 and 4. Tissue becomes darker in color.
7. Tone in Gold Chloride Solution for 20 to 30 minutes.
8. Place sections directly into Sodium Thiosulfate, 5% for 1 minute. Rinse in distilled water.
9. Dehydrate in 95% alcohol, absolute alcohol and clear in Xylene two changes each.
10. Mount with Permount or any synthetic mounting medium

Stain Results:  

Various lipid granules are unstained, but each granule is clearly outlined.

References:

AFIP, Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna: New York: McGraw-Hill Publications. C. 1968, p. 198.

Hirano, A. and Zimmerman, H.M., Arch. Neurol., 6:114-122, 1962.