Sweat, Meloan & Puchtler's
One Step Trichrome

Solutions

Solution A
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
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.

Method

1. Bring sections to water via xylene and ethanol.
2. Place in Bouin's fixative at 56°C for one hour.
3. Wash with running tap water 3 to 4 minutes to remove yellow.
4. Place into solution A for 1 minute.
5. Rinse with solution B for 30 seconds.
6. Dehydrate rapidly with ethanol.
7. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei, cytoplasm, muscle, fibrin & elastic  –  red
• Collagen  –  blue

Notes

1. A nuclear stain is not used as the hydrochloric acid will remove it.

 

Reference

Culling, C.F.A., Alison, R.T. and Barr, W.T. (1985)
Cellular Pathology Technique, 4th ed.
Butterworths, London, UK.

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