Bielschowsky Silver Tissue Stain for Axons and Alzheimer’s Disease

Contents

  1. Materials
  2. Procedure
  3. Results
  • Acknowledgements

    • 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

    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.

    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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