Mollier's trichrome
for Elastic and Collagen

Solutions

Weigert's iron hematoxylin or equivalent

Solution A
Orcein	0.8	g
Hydrochloric acid	1	mL
Ethanol, 100%	50	mL
Distilled water	50	mL

Solution B
Azocarmine	2	g
Acetic acid, glacial	1	mL
Distilled water	100	mL

Solution C
Phosphotungstic acid	5	g
Distilled water	100	mL

Solution D
Naphthol green B	1	g
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 into solution A for 12 hours.
3. Stain nuclei with Weigert's iron hematoxylin for 1-3 minutes.
4. Differentiate the nuclear stain if necessary.
5. Wash with water for 15 minutes.
6. Place into solution B for 15-30 minutes.
7. Rinse with distilled water.
8. Decolourise in solution C for 2-6 hours, changing the solution three times.
9. Rinse quickly with distilled water.
10. Place into solution D for 15-30 minutes.
11. Agitate vigorously in 95% ethanol for 30 seconds.
12. Dehydrate with ethanol.
13. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Elastic  –  black
• Erythrocytes  –  red
• Cytoplasm  –  purple
• Collagen  –  green

Notes

1. Solution A is the Unna-Taenzer elastic solution.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Mollier, (1938)
    Zeitschrift für wissenschaftliche Mikroskopie und für mikroskopische Technik,
    v.55, pp.472.
  And:–
    von Kahlden, C. and Laurent, O., (1896)
    Technique microscopique, pp.143
    Carré, Paris, France