Congo Red Stain Protocol For Amyloid

Run a control slide.                                     Control tissue – Tissue with amyloid

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. 1% Congo red ————————————— 60 minutes (re-use)
  8. Rinse in water ————————————— 1 minute
  9. Alkaline Alcohol ————————————- until background clear (re-use)
  10. Wash in running water —————————- 5 minutes
  11. Harris Hematoxylin ——————————— 30 seconds (re-use)
  12. Wash in running water —————————- 10 minutes
  13. Dehydrate in 95% alcohol———————— 30 seconds
  14. Dehydrate in 100% alcohol——————— 1 minute
  15. Dehydrate in 100% alcohol——————— 2 minutes
  16. Dehydrate in 100% alcohol——————— 2 minutes
  17. Clear in xylene (3)——————————– 2 minutes (re-use)
  18. Clear in xylene (4)——————————– 5 minutes (re-use)
  19. Coverslip

Results:         Amyloid – Pink to red                     Nuclei – Blue

With polarizing microscope

Amyloid – Green Birefringence

 

An Expert Histology Service


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