Wallart and Honette's Trichrome
for Connective Tissues

Solutions

Stock solution 1
Acid fuchsin	1	g
Acetic acid	1	mL
Distilled water	100	mL
Stock solution 2
Fast yellow	3	g
Acetic acid	1	mL
Distilled water	100	mL
Stock solution 3
Phosphomolybdic acid	1	g
Distilled water	100	mL

Working solution A
Stock solution 1	30	mL
Stock solution 2	30	mL
Stock solution 3	30	mL
Working solution B
Acetic acid	1	mL
Distilled water	100	mL
Working solution C
Acetic acid	1	mL
Ethanol 100%	100	mL

Tissue sample
5µ paraffin sections of formalin fixed tissue are suitable. Many 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 an acid resistant nuclear stain.
3. Rinse well with tap water.
4. Place into solution A for 5 minute.
5. Rinse quickly with distilled water.
6. Place into solution B for 5 minutes.
7. With a pipette, drop solution C onto the slide for 30 seconds.
8. Dehydrate with ethanol.
9. Clear with xylene and mount with a resinous medium.

Expected results

• Elastic  –  pink
• Cytoplasm  –  red
• Collagen  –  yellow
• Nuclei  –  black

Reference
Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Wallart and Honette, (1934)
    Bulletin d'histologie appliquée à la physiologie et à la pathologie
    et de technique microscopique, vol. 10, pp. 404