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:  

Nucleolus and Nuclear Membrane

Black

Neurofibrils, Dendrites and Axis Cylinders

Black

Cytoplasm of Astrocytes and the Cytoplasmic membrane of Macrophages

Grey

Senile Plaques

Black

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.