Bielschowsky Silver Tissue Stain for Axons and Alzheimer’s Disease

The Bielschowsky Silver Stain is a classical histological silver stain used for the impregnation of nerve fibers and the detection of neurites and neurofibrillary tangles associated with Alzheimer’s disease, yielding a black stain.


8 micron paraffin wax sections
Solution A: 20% silver nitrate in distilled water

Reducing Solution (make fresh just before use)

  • Formaldehyde 50ml
  • Absolute Alcohol 200ml
  • Distilled water 750ml
Solution B
  • 20% Silver nitrate 20ml
  • 95% Alcohol 20ml
  • Add ammonia until precipitate formed is redissolved
  • Add 10 more drops of ammonia

5% Sodium thiosulphate
0.1% Gold chloride


  1. Take sections to distilled water
  2. Place in solution A for 2-3 hours
  3. Wash in reducer for 2-5 minutes
  4. Wash in 10% alcohol for 2 minutes
  5. Place in solution B for 1-2 minutes
  6. Wash in a constant flow of reducer and leave for 3-5 minutes
  7. Wash in distilled water for 2 minutes.
  8. Tone 0.1% gold chloride for 2 minutes
  9. Wash in distilled water for 2 minutes.
  10. Fix in 5% sodium thiosulphate for 2 minutes
  11. Wash in tap water for 5 minutes.
  12. Dehydrate through 70, 80, 90, and two changes of 100% ethanol.
  13. Clear by immersing slides in two changes of xylene solutions for 5 minutes each.


Very high magnification micrograph of the cerebellum (cerebellar cortex). Bielschowsky stain. Source: User: Nephron (Wikipedia)


This protocol is based upon a published procedure by Jim Lowe at the University of Nottingham.

Comments are closed.