Sweat, Meloan & Puchtler's
One Step Trichrome
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Chromotrope 2R | 0.6 | g |
| Aniline blue | 0.6 | g |
| Phosphomolybdic acid | 1 | g |
| Hydrochloric acid, conc. | 1 | mL |
| Distilled water | 100 | mL |
Solution B
| Material | Amount | |
|---|---|---|
| Acetic acid, glacial | 1 | 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.
- Place in Bouin’s fixative at 56°C for one hour.
- Wash with running tap water 3 to 4 minutes to remove yellow.
- Place into solution A for 1 minute.
- Rinse with solution B for 30 seconds.
- Dehydrate rapidly with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei, cytoplasm, muscle, fibrin & elastic – red
- Collagen – blue
Notes
- A nuclear stain is not used as the hydrochloric acid will remove it.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling, C.F.A., Alison, R.T. and Barr, W.T. (1985)
Cellular Pathology Technique, 4th ed.
Butterworths, London, UK.
