Boone & Drijver's
Alcoholic Trichrome for Exfoliated Cells


Mayer's hemalum or equivalent
Solution A
Orange G 1 g
Phosphotungstic acid 0.075 g
Ethanol, absolute 250 mL
Distilled water 250 mL
Combine the ethanol and water. Add the orange G and phosphotungstic acid. Filter. The pH should be between 2.0 and 2.8.
Solution B
Eosin Y 0.9 g
Light green SF 0.35 g
Phosphotungstic acid   1.5 g
Ethanol, 100% 250 mL
Distilled water 250 mL
Lithium carbonate as needed  
Combine the ethanol and water. Dissolve the dyes, then the acid. Stirring continuously, adjust to pH4.6 with lithium carbonate powder. Filter.

Tissue sample
Ethanol fixed smears of exfoliated epithelial cells.


  1. Bring smears to water via graded ethanols.
  2. Stain nuclei with alum hematoxylin for 2 minutes.
  3. Blue with running tap water.
  4. Rinse with 50% ethanol, 15 dips.
  5. Place into solution A for 1 minute.
  6. Rinse with 50% ethanol, two changes of 15 dips each.
  7. Place into solution B for 2 minutes.
  8. Rinse with 50% ethanol, 15 dips.
  9. Dehydrate with absolute ethanol, two changes.
  10. Clear with t-butanol and mount with a resinous medium.

Expected results


Kiernan. J.A., (1999)
Histological and histochemical methods: Theory and practice, Ed. 3
Butterworth Heinemann, Oxford, UK.
    Boone, M. E., and Drijver, J. S., (1986)
    Routine cytological staining. Theoretical background and practice.
    Elsevier, New York.




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