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Goldner’s Trichrome for Muscle and Collagen

Protocol

Goldner's Trichrome

for Muscle and Collagen

11
steps
9
materials

Materials

  • Weigert’s iron hematoxylin or equivalent
  • Solution A
    MaterialVariation 1Variation 2Variation 3
    Ponceau 2R0.7g0.75g
    Acid fuchsin0.3g0.25g
    Azophloxine0.1g5g
    Acetic acid, glacial2mL2mL2mL
    Distilled water1L1L1L
  • Solution B
    MaterialAmount
    Acetic acid, glacial10mL
    Distilled water1L
  • Solution C
    MaterialAmount
    Orange G20g
    Phosphomolybdic acid40g
    Distilled water1L
  • Solution D
    MaterialAmount
    Light green SF2g
    Acetic acid, glacial2mL
    Distilled water1L

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.


Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Stain nuclei with Weigert’s iron hematoxylin, or equivalent.
  3. Place into one of the variants of solution A for 5 minutes.
  4. Rinse with solution B.
  5. Place into solution C until collagen is decolorised.
  6. Rinse with solution B.
  7. Place into solution D for 5 minutes.
  8. Place into solution B for 5 minutes.
  9. Blot.
  10. Dehydrate rapidly with absolute ethanol.
  11. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Erythrocytes  –  orange
  • Cytoplasm  –  red
  • Collagen  –  green

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.