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Pollak’s Trichrome for Muscle and Collagen


Pollak's Trichrome

for Muscle and Collagen



  • Weigert’s iron hematoxylin or equivalent
  • Solution A
    Orange G0.25g
    Ponceau 2R0.33g
    Acid fuchsin0.17g
    Light green SF0.15g
    Phosphotungstic acid0.5g
    Phosphomolybdic acid0.5g
    Acetic acid, glacial1mL
    Ethanol, 95%50mL
    Distilled water50mL
  • Solution B
    Acetic acid, glacial0.2mL
    Distilled water100mL

Preparation of Solution A

  1. Mix the water, ethanol and acetic acid together.
  2. Divide into four, and dissolve as follows:
    1. Phosphomolybdic acid
    2. Phosphotungstic acid and orange G
    3. Light green SF
    4. Ponceau 2R and acid fuchsin
  3. Combine all solutions and filter.

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 Weigert’s iron hematoxylin or equivalent.
  3. Place into solution A for 3-7 minute.
  4. Place into solution B until differentiated.
  5. Dehydrate with ethanol.
  6. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Erythrocytes  –  yellow
  • Cytoplasm  –  red
  • Muscle  –  red
  • Collagen  –  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.
    Pollack, (1944)
    Archives of pathology and laboratory medicine, v. 37, pp. 294
    Chicago, USA