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Möllendorf’s trichrome for Muscle and Collagen


Möllendorf's trichrome

for Muscle and Collagen



  • Hansen’s iron hematoxylin or equivalent
  • Solution A
    Eosin Y1g
    Acetic acid, glacial0.3mL
    Distilled water100mL
  • Solution B
    Phosphomolybdic acid2g
    Distilled water100mL
  • Solution C
    Methyl blue1g
    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. Stain nuclei with Hansen’s iron hematoxylin for 5 minutes.
  3. Rinse with distilled water.
  4. Wash with running tap water.
  5. Place into solution A for 20 minute.
  6. Rinse with distilled water.
  7. Place into solution B for 10 seconds.
  8. Rinse with distilled water.
  9. Place into solution C for 1-2 minutes.
  10. Rinse with distilled water.
  11. Place into 95% ethanol until clouds of dye cease to flood away.
  12. Dehydrate with ethanol.
  13. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Cytoplasm  –  pink
  • Muscle  –  pink
  • Collage  –  blue

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.
    Roskin, G.E., (1946)
    Mikroskopecheskaya technika, pp.158
    Moscow, Sovetskaya Nauka.