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