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Kostowiecki’s Trichrome for Muscle, Cartilage and Collagen


Kostowiecki's Trichrome

for Muscle, Cartilage and Collagen



  • A red nuclear stain
  • Solution A
    Aniline blue0.06g
    Orange G0.2g
    Phosphomolybdic acid1g
    Distilled water100mL

Preparation of Solution A

  1. Add both dyes to the water.
  2. Bring to the boil for three minutes.
  3. Add the phosphomolybdic acid while hot.
  4. Cool 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 a red nuclear stain.
  3. Place into the staining solution until darkly stained, 30-120 minutes.
  4. Rinse with distilled water.
  5. Place in 95% ethanol for 1 minute.
  6. Dehydrate with absolute ethanol.
  7. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  red
  • Muscle  –  orange
  • Cartilage  –  blue
  • Collagen  –  blue-green

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.
    Kostowiecki, (1932)
    Zeitschrift für wissenschaftliche Mikroskopie und für mikroskopische Technik,
    v. 49, pp. 337