Safranin O Fast green Stain Protocol

Rat Joint Safranin O

Anyone out there doing articular cartilage studies?  This stain is for you.  A fast easy stain that can give beautiful results.  I wish all stains were this easy.  Take a look.

Control tissue:  Use bone with joint attached.

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. Harris Hematoxylin—————————-  5 minutes (re-use)
  8. Define in 1% acid alcohol———————— 1 dip (re-use)
  9. Blue in ammonium water———————–  30 seconds (re-use)
  10. 10.  Rinse in water—————————— 1 minute
  11. Stain in Fast green—————————— 5 minutes (re-use)
  12. Rinse in water———————————-  30 seconds
  13. 1% acetic acid———————————– 30 seconds (re-use)
  14. Stain in Safranin O—————————— 15 minutes (re-use)
  15. Rinse in water———————————- 30 seconds
  16. Dehydrate in 95% alcohol———————– 30 seconds
  17. Dehydrate in 100% alcohol———————- 1 minute
  18. Dehydrate in 100% alcohol———————- 2 minutes
  19. Dehydrate in 100% alcohol———————- 2 minutes
  20. Clear in xylene (3)—————————— 2 minutes (re-use)
  21. Clear in xylene (4)—————————— 5 minutes (re-use)
  22. Coverslip

Results:

Articular cartilage (proteoglycan) – Red

Nuclei- Blue

Background- Green

 

An Expert Histology Service

4 thoughts on “Safranin O Fast green Stain Protocol

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