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Brillmeyer’s Trichrome for Muscle and Collagen

Brillmeyer's Trichrome

for Muscle and Collagen

8
steps
6
materials

Materials

Solution A

MaterialAmount
Acid fuchsin0.2g
Distilled water100mL

Solution B

MaterialAmount
Aniline blue0.5g
Orange G2g
Phosphomolybdic 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.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Stain with an acid resistant nuclear stain.
  3. Place into solution A for 1 minute.
  4. Drain.
  5. Place into solution B for 2-3 hours.
  6. Wash briefly with distilled water.
  7. Dehydrate with ethanol.
  8. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue or black
  • Erythrocytes  –  yellow
  • Cytoplasm  –  red
  • Collagen  –  blue

Notes

  • The original technique recommended nuclear staining in Delafield’s alum hematoxylin, with blueing.
  • Weiss modified this method to shorten the time required.

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.