Orcein for Elastic Fibers Stain Protocol

This elastic stain is far easier than Verhoeff Van Gieson stain but the chemical can be more unusual to find.

Run a control slide.             Control Tissue – Aorta, Skin

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. Stain in Orcein ————————————– 30 minutes (re-use)
  8. 70% alcohol —————————————– 10 minutes
  9. 70% alcohol —————————————– 10 minutes
  10. 10.  70% alcohol —————————————- 10 minutes
  11. 11.  Rinse in running water ————————– 1 minute
  12. 12.  Stain in Harris Hematoxylin ——————– 1 minute (re-use)
  13. 13.  Rinse in running water ————————– 1 minute
  14. 14.  Acid alcohol —————————————- 1 dip (re-use)
  15. 15.  Rinse in running water ————————– 5 minutes
  16. 16.  Dehydrate in 95% alcohol———————- 30 seconds
  17. 17.   Dehydrate in 100% alcohol——————- 1 minute
  18. 18.   Dehydrate in 100% alcohol——————- 2 minutes
  19. 19.   Dehydrate in 100% alcohol——————- 2 minutes
  20. 20.   Clear in xylene (3)—————————— 2 minutes (re-use)
  21. 21.   Clear in xylene (4)—————————— 5 minutes (re-use)

Results:         Elastic Fibers – Dark Pink

 

 

An Expert Histology Service


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