Safranin O Fast green Stain Protocol

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


Articular cartilage (proteoglycan) – Red

Nuclei- Blue

Background- Green


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We are in the business of histology to help researchers and patients get the fast accurate results they deserve. Most of our techniques are old school protocols that have been modified through hard work and persistence. Call or email us today and see what a difference we can make for you.
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4 Responses to Safranin O Fast green Stain Protocol

  1. Everything is very open with a very clear description of the
    issues. It was really informative. Your website is extremely
    helpful. Thanks for sharing!


  2. Its such as you read my mind! You appear to know a lot approximately this, such as you wrote the ebook in it or something.
    I feel that you just could do with some p.c. to drive the message home a bit,
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  3. Kathleen Roberts says:

    A possibly dumb question, but what is the concentration of Safranin O and Fast Green?

    Liked by 1 person

  4. They are both 0.1% solutions.


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