This is a great stain if you have the controls to run.
Safety equipment: Work under a hood with lab coat, gloves, and glasses.
- De-paraffin slides in xylene (1) ———————- 2 minutes (re-use)
- De-paraffin slides in xylene (2)———————– 2 minutes (re-use)
- Clear slides in 100% alcohol————————– 2 minutes
- Clear slides in 100% alcohol————————– 2 minutes
- Hydrate slides in 95% alcohol———————— 2 minutes
- Hydrate slides in running water———————- 5 minutes
- Uranyl nitrate at 60C ——————————– 15 minutes
- Rinse in running water ——————————- 1 minute
- 1% silver nitrate at 60C —————————— 90 minutes
10. Rinse in running water —————————— 1 minute
11. Dehydrate in 95% alcohol —————————- 1 minute
12. Dehydrate in 100% alcohol ————————– 1 minute
13. 2.5% gum mastic ———————————— 5 minutes
14. Air dry ———————————————- 5 minutes
15. Place in reducing solution at 45C ——————— 25 minutes
16. Check with microscope, spirochetes should be black.
17. If not dark enough, place in reducing solution at room temp for 1-2 minutes
18. Check again with microscope
19. Dehydrate in 95% alcohol———————— 30 seconds
20. Dehydrate in 100% alcohol———————- 1 minute
21. Dehydrate in 100% alcohol———————- 2 minutes
22. Dehydrate in 100% alcohol———————- 2 minutes
23. Clear in xylene (3)——————————- 2 minutes (re-use)
24. Clear in xylene (4)——————————- 5 minutes (re-use)
25. Coverslip
Results: Spirochetes – Black
Background – Yellowish
An Expert Histology Service
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