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Patay’s Trichrome for Muscle and Collagen

Patay's Trichrome

for Muscle and Collagen



  • Masson’s alum hematoxylin
  • Solution A
    Ponceau 2R1g
    Distilled water100mL
  • Solution B
    Phosphomolybdic acid1g
    Distilled water100mL
  • Solution C
    Light green SF0.5g
    Ethanol, 90%100mL

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. Stain nuclei with Masson’s alum hematoxylin, underdifferentiate and blue.
  3. Place into solution A for 2 minute.
  4. Rinse with distilled water.
  5. Place into solution B for 2 minutes.
  6. Rinse quickly with distilled water.
  7. Place into solution C for 30 seconds.
  8. Dehydrate with ethanol.
  9. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue
  • Cartilage  –  blue
  • Erythrocytes  –  yellow
  • Cytoplasm  –  orange
  • Muscle  –  orange
  • Bone  –  bright green
  • Collagen  –  green


  • Gray strongly recommended this technique. He considered it to be the finest trichrome described to that date (1954).
  • The blue of the cartilage is from using a ripened, strong hemalum.

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.
    Patay, (1934)
    Bulletin d’histologie appliquée à la physologie et à la pathologie et de technique microscopique, v.11, pp.408