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Electron Microscopy Sciences

Technical Data Sheets

Hirano-Zimmerman for Nerve Cells & Fibers

EMS Catalog #: 26698-Series

Fixation:

Formalin 10% Neutral Buffered (#15740).

Sections:

Paraffin @ 12 – 15 microns

Staining Procedures:

  1. Deparaffinize and hydrate to distilled water.
  2. Silver Nitrate, 10% (#26698-01) for two hours or longer.
  3. Place sections directly in Ammonia Water (#26698-02) for 3 minutes.
  4. Place section directly in Formalin Solution (#26698-03) for 3 minutes. Tissue becomes black-brown. Rinse in distilled water, two changes.
  5. Place section again into Silver Nitrate, 10% (#26698-01) for 3 – 5 minutes.
  6. Repeat step 3 and 4. Tissue becomes darker in color.
  7. Tone in Gold Chloride Solution (#26698-04) for 20 to 30 minutes.
  8. Place sections directly into Sodium Thiosulfate, 5% (#26698-05) for 1 minute. Rinse in distilled water.
  9. Dehydrate in 95% alcohol, absolute alcohol and clear in Xylene (#23400) two changes each.
  10. Mount with Permount (#17986-01) 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 Gray
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.

Online Ordering

Hirano-Zimmerman for Nerve Cells & Fibers is available online from the EMS Catalog. For ordering or product information, click here.