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Roque’s Trichrome for Mallory Bodies


Roque's Trichrome

for Mallory Bodies



  • Weigert’s iron hematoxylin or equivalent
  • Solution A
    Phosphomolybdic acid1g
    Distilled water100mL
  • Solution B
    Aniline blue0.5g
    Chromotrope 2R2g
    Hydrochloric acid, 0.02N100mL

    Dissolve the aniline blue in the acid with gentle heat. Cool. Add the chromotrope 2R. Filter.

Tissue Sample

3µ paraffin sections of neutral buffered formalin fixed liver are suitable. Other fixatives are likely to be satisfactory. Most trichrome stains benefit from picric acid or mercuric chloride fixation. Formalin fixed tissues may benefit from secondary fixation of sections in Bouin’s fluid.


  1. Bring sections to water via xylene and ethanol.
  2. Stain nuclei with Weigert’s iron hematoxylin or equivalent for 5 minutes.
  3. Wash in tap water to blue.
  4. Place into solution A for 2 minutes.
  5. Rinse with distilled water.
  6. Place into solution B for 8 minutes.
  7. Rinse with distilled water.
  8. Dehydrate with ethanol.
  9. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Cytoplasm  –  red
  • Muscle  –  red
  • Mallory bodies  –  bright blue, or red with blue periphery
  • Collagen  –  blue

Safety Note

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


  1. Roque, Agustin L., (1953),
    Chromotrope aniline blue method of staining Mallory bodies of Laennec’s cirrhosis.
    Laboratory Investigation, v. 2, no. 5, pp. 15-21.