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Papanicolaou Stain–Harris

Reagent Preparation

Mix one part Scott’s Tap Water Substitute with 9 parts deionized water (i.e. one bottle with 900 mL deionized water).


  1. Fix slides in 95% alcohol for 15 minutes.
  2. Rinse in tap water.
  3. Stain in Harris’ Hematoxylin solution for 1 to 3 minutes.
  4. Rinse in tap water.
  5. Dip slides in Differentiating solution for 20 to 60 seconds.
  6. Rinse in tap water.
  7. Dip slides 10 times in working Scott’s Tap Water Substitute.
  8. Rinse in tap water.
  9. Dip slides 10 times in 95% alcohol.
  10. Stain in OG-6 Solution for 1.5 minutes.
  11. Dip slides 10 times in 95% alcohol.
  12. Stain in EA-50 Solution for 2.5 minutes.
  13. Dehydrate in 2 changes of 95% alcohol.
  14. Dehydrate in 100% alcohol for 1 minute.
  15. Clear in 2 changes of xylene and coverslip with mounting media.


Nuclei are stained blue while cytoplasm displays varying shades of blue, orange, pink, and red.

Papanicolaou Stain-Harris


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