Bielschowsky for Neurofibrils and Senile Plaques (Alzheimer)

In addition to the Tau and Amyloid proteins discovered through IHC, this silver stain indicates Alzheimer disease.

Control tissue: Brain Cortex, Spinal Cord

Safety equipment: Work under a hood with lab coat, gloves, and glasses.

  1. De-paraffin slides in xylene (1) —————— 2 minutes (re-use)
  2. De-paraffin slides in xylene (2)——————- 2 minutes (re-use)
  3. Clear slides in 100% alcohol———————- 2 minutes
  4. Clear slides in 100% alcohol———————- 2 minutes
  5. Hydrate slides in 95% alcohol——————– 2 minutes
  6. Hydrate slides in running water —————– 5 minutes
  7. 10% Silver nitrate ——————————  30 minutes in 40C oven
  8. Place slides in DH2O—————————-  until new solution is made
  9. Slowly add ammonium hydroxide to the 10% silver nitrate until it is clear.
  10. Place slides in solution in 40C oven ————– 30 minutes
  11. Place in developer solution ———————- 5 minutes (re-use)
  12. Slides should be golden, use scope to check.
  13. Place in 1% ammonium hydroxide to stop reaction – 1 minute
  14. Wash in water ———————————– 2-5 minutes
  15. Place in 5% sodium thiosulfate ——————- 5 minutes (re-use)
  16. Wash in running water ————————–  5 minutes
  17. Dehydrate in 95% alcohol———————— 30 seconds
  18. Dehydrate in 100% alcohol———————–  1 minute
  19. Dehydrate in 100% alcohol———————–  2 minutes
  20. Dehydrate in 100% alcohol———————–  2 minutes
  21. Clear in xylene (3)——————————-  2 minutes (re-use)
  22. Clear in xylene (4)——————————-  5 minutes (re-use)

Results:           Neurofibrils and senile plaques- black

Background- yellow to brown

 

An Expert Histology Service


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