Protocol
Roque's Trichrome
for Mallory Bodies
Materials
- Weigert’s iron hematoxylin or equivalent
- Solution A
Material Amount Phosphomolybdic acid 1 g Distilled water 100 mL - Solution B
Material Amount Aniline blue 0.5 g Chromotrope 2R 2 g Hydrochloric acid, 0.02N 100 mL 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.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with Weigert’s iron hematoxylin or equivalent for 5 minutes.
- Wash in tap water to blue.
- Place into solution A for 2 minutes.
- Rinse with distilled water.
- Place into solution B for 8 minutes.
- Rinse with distilled water.
- Dehydrate with ethanol.
- 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.
References
- 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.
