Steiner and Steiner for Spirochetes Stain Protocol

This is a great stain if you have the controls to run.

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. Uranyl nitrate at 60C ——————————– 15 minutes
  8. Rinse in running water ——————————- 1 minute
  9. 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


One thought on “Steiner and Steiner for Spirochetes Stain Protocol

  1. An intriguing discussion is definitely worth comment. I do think that you should publish more about this subject, it might not be a taboo subject but typically people do not speak about these subjects. To the next! Many thanks!!

    Like

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