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Boone & Drijver’s Alcoholic Trichrome for Exfoliated Cells

Protocol

Boone & Drijver's Alcoholic Trichrome

for Exfoliated Cells

10
steps
11
materials

Materials

  • Mayer’s hemalum or equivalent
  • Solution A
    MaterialAmount
    Orange G1g
    Phosphotungstic acid0.075g
    Ethanol, absolute250mL
    Distilled water250mL

    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
    MaterialAmount
    Eosin Y0.9g
    Light green SF0.35g
    Phosphotungstic acid1.5g
    Ethanol, 100%250mL
    Distilled water250mL
    Lithium carbonateas needed

Tissue Sample

Ethanol fixed smears of exfoliated epithelial cells.


Protocol

  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

  • Nuclei  –  blue
  • Cytoplasm  –  pink or blue-green
  • Keratinized cells  –  orange
  • Erythrocytes  –  pink to orange

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.


References

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