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

Very high magnification micrograph of the cerebellum (cerebellar cortex). Bielschowsky stain. Source: User: Nephron (Wikipedia)
Acknowledgements
This protocol is based upon a published procedure by Jim Lowe at the University of Nottingham.
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