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Lee-Brown’s Trichrome for Muscle and Collagen

Lee-Brown's Trichrome

for Muscle and Collagen


This is a modification of Mallory’s trichrome.


Solution A

Acid fuchsin1g
Distilled water100mL

Solution B

Phosphomolybdic acid1g
Distilled water100mL

Solution C

Orange G2g
Methyl blue0.5g
Oxalic acid2g
Distilled water100mL

Tissue Sample

5µ paraffin sections of neutral buffered formalin fixed tissue 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. Place into solution A for 2 minute.
  3. Rinse with distilled water.
  4. Place into solution C for 15 minutes.
  5. Rinse quickly with distilled water.
  6. Place into solution B for 2 minutes.
  7. Wash well with distilled water.
  8. Dehydrate and differentiate with ethanol.
  9. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  red
  • Erythrocytes  –  orange
  • Keratin  –  orange
  • Muscle  –  red
  • Collagen  –  blue


  • This modification reverses the staining sequence of Mallory’s original method, and applies solution C before solution B.
  • Although an acid resistant nuclear stain is not specified, Weigert’s iron hematoxylin or an equivalent could be done before step 2. The nuclei would then be black.

Safety Note

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


  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Mallory, (1901)
    Journal of Experimental Medicine, v.5, pp.15
    Lee-Brown, (1929)
    Journal of Urology, v.21, pp.259