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Cason’s Trichrome for Muscle and Collagen

Cason's Trichrome

for Muscle and Collagen

5
steps
5
materials

Materials

Staining solution

MaterialAmount
Orange G1g
Acid fuchsin1.5g
Aniline blue0.5g
Phosphotungstic acid1g
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.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Place into the staining solution for 5 minute.
  3. Rinse with distilled water.
  4. Dehydrate with ethanol.
  5. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  red
  • Erythrocytes  –  orange
  • Cytoplasm  –  red
  • Collagen  –  blue

Notes

  • Although the method does not specifically call for one, an acid resistant nuclear stain could be inserted immediately before step 2.

Safety Note

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

References

  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Citing:
    Cason, (1950)
    Stain technology, vol.25, pp.225