Kricheski's Trichrome
for Muscle and Collagen
Materials
Solution A
Material | Amount | |
---|---|---|
Acid fuchsin | 0.25 | g |
Distilled water | 100 | mL |
Solution B
Material | Amount | |
---|---|---|
Methyl blue, 1% aqueous | 30 | mL |
Orange G, 1% aqueous | 30 | mL |
Phosphomolybdic acid, 1% aqueous | 30 | mL |
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
- Bring sections to water via xylene and ethanol.
- Place into solution A for 1-3 minute.
- Rinse well with distilled water.
- Place into solution B for 3-5 minutes.
- Dip 2 or 3 times in 70% ethanol.
- Dehydrate and differentiate with ethanol for 1-3 minutes.
- Clear with xylene and mount with a resinous medium.
- 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
- 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