Protocol
Möllendorf's trichrome
for Muscle and Collagen
Materials
- Hansen’s iron hematoxylin or equivalent
- Solution A
Material Amount Eosin Y 1 g Acetic acid, glacial 0.3 mL Distilled water 100 mL - Solution B
Material Amount Phosphomolybdic acid 2 g Distilled water 100 mL - Solution C
Material Amount Methyl blue 1 g 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 Hansen’s iron hematoxylin for 5 minutes.
- Rinse with distilled water.
- Wash with running tap water.
- Place into solution A for 20 minute.
- Rinse with distilled water.
- Place into solution B for 10 seconds.
- Rinse with distilled water.
- Place into solution C for 1-2 minutes.
- Rinse with distilled water.
- Place into 95% ethanol until clouds of dye cease to flood away.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Cytoplasm – pink
- Muscle – pink
- Collage – blue
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:
Roskin, G.E., (1946)
Mikroskopecheskaya technika, pp.158
Moscow, Sovetskaya Nauka.
