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Kricheski’s Trichrome for Muscle and Collagen

Kricheski's Trichrome

for Muscle and Collagen

7
steps
5
materials

Materials

Solution A

MaterialAmount
Acid fuchsin0.25g
Distilled water100mL

Solution B

MaterialAmount
Methyl blue, 1% aqueous30mL
Orange G, 1% aqueous30mL
Phosphomolybdic acid, 1% aqueous30mL

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 solution A for 1-3 minute.
  3. Rinse well with distilled water.
  4. Place into solution B for 3-5 minutes.
  5. Dip 2 or 3 times in 70% ethanol.
  6. Dehydrate and differentiate with ethanol for 1-3 minutes.
  7. Clear with xylene and mount with a resinous medium.
  8. Bring sections to water via xylene and ethanol.

Expected Results

  • Nuclei – red
  • Erythrocytes – orange
  • Keratin – orange
  • Muscle – red
  • Collagen – blue

Notes

  • Although this method does not specify that an acid resistant nuclear stain be used, Weigert’s iron hematoxylin or equivalent could be inserted before step 2, lightly differentiated and blued. The nuclei would then be black.

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:
    Kricheski, (1931)
    Stain technology, v.6, pp.97