Papanicolaou's Trichrome
for Exfoliated Cells

This is not the Papanicolaou technique commonly used to screen for cervical cancer.


Ehrlich's alum hematoxylin or equivalent
Staining solution
Acid fuchsin 1 g
Aniline blue 0.6 g
Eosin Y 2.1 g
Orange G 1.25 g
Phosphotungstic acid 1.12
Phosphomolybdic acid   2.25 g
Distilled water 1 L

Tissue sample
Ethanol fixed smears of exfoliated epithelial cells.


  1. Stain nuclei with Ehrlich's hematoxylin for 2 minutes.
  2. Blue.
  3. Rinse with distilled water.
  4. Place into the staining solution for 2-5 minutes.
  5. Rinse quickly with distilled water.
  6. Dehydrate rapidly with ethanol.
  7. Clear with xylene and mount with a resinous medium.

Expected results


  1. The method specified that dehydration and clearing should be with dioxane. This reagent is no longer used due to its toxicity. If satisfactory results are not obtained with rapid ethanol dehydration, dehydrants such as t-butanol should be tried.


Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
    Papanicolaou, (1941)
    Journal of laboratory and clinical medicine, v. 26, pp. 1200




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