Patay's Trichrome
for Muscle and Collagen

Solutions

Masson's alum hematoxylin

Solution A
Ponceau 2R	1	g
Distilled water	100	mL

Solution B
Phosphomolybdic acid	1	g
Distilled water	100	mL

Solution C
Light green SF	0.5	g
Ethanol, 90%	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. Stain nuclei with Masson's alum hematoxylin, underdifferentiate and blue.
3. Place into solution A for 2 minute.
4. Rinse with distilled water.
5. Place into solution B for 2 minutes.
6. Rinse quickly with distilled water.
7. Place into solution C for 30 seconds.
8. Dehydrate with ethanol.
9. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Cartilage  –  blue
• Erythrocytes  –  yellow
• Cytoplasm  –  orange
• Muscle  –  orange
• Bone  –  bright green
• Collagen  –  green

Notes

1. Gray strongly recommended this technique. He considered it to be the finest trichrome described to that date (1954).
2. The blue of the cartilage is from hematoxylin.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Patay, (1934)
    Bulletin d'histologie appliquée à la physologie et à la pathologie et de technique
    microscopique, v.11, pp.408