Protocol
Goldner's Trichrome
for Muscle and Collagen
Materials
- Weigert’s iron hematoxylin or equivalent
- Solution A
Material Variation 1 Variation 2 Variation 3 Ponceau 2R 0.7 g 0.75 g – Acid fuchsin 0.3 g 0.25 g – Azophloxine – 0.1 g 5 g Acetic acid, glacial 2 mL 2 mL 2 mL Distilled water 1 L 1 L 1 L - Solution B
Material Amount Acetic acid, glacial 10 mL Distilled water 1 L - Solution C
Material Amount Orange G 20 g Phosphomolybdic acid 40 g Distilled water 1 L - Solution D
Material Amount Light green SF 2 g Acetic acid, glacial 2 mL Distilled water 1 L
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
- Bring sections to water via xylene and ethanol.
- Stain nuclei with Weigert’s iron hematoxylin, or equivalent.
- Place into one of the variants of solution A for 5 minutes.
- Rinse with solution B.
- Place into solution C until collagen is decolorised.
- Rinse with solution B.
- Place into solution D for 5 minutes.
- Place into solution B for 5 minutes.
- Blot.
- Dehydrate rapidly with absolute ethanol.
- 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
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.