Gomori's Trichrome
for Muscle and Collagen
Materials
- Weigert’s iron hematoxylin or equivalent
- Solution A
Material Amount Chromotrope 2R 0.6 g Fast green FCF 0.3 g Phosphotungstic acid 0.6 g Acetic acid, glacial 1 mL Distilled water 99 mL - Solution B
Material Amount Acetic acid, glacial 0.2 mL Distilled water 100 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.
- Stain nuclei with an acid resistant nuclear stain.
- Place into solution A for 5-20 minute.
- Quickly rinse off stain with distilled water.
- Rinse with solution B.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Cytoplasm – red
- Collagen – green
Notes
- The time in solution A should be determined empirically. Once a satisfactory stain has been achieved, the time should remain constant. Any change in fixation, processing or section thickness warrants reviewing the staining time.
- Light green SF yellowish can replace Fast green FCF.
- Wheatley recommended this trichrome for protozoa in intestinal smears.
- Engel & Cunningham modified this method to differentiate muscle fibre types in cryostat sections.
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:
Gomori, (1950),
Technical Bulletin, vol.20, pp.77
And:
Wheatley, (1951)
Technical Bulletin, vol.21, pp.92 - Humason, G. L., (1967)
Animal tissue techniques, Ed. 2
W. H. Freeman and Company, San Francisco, Ca, USA
