Protocol
Wallart & Honette's Trichrome
for Connective Tissues
Materials
Stock solution 1
| Material | Amount | |
|---|---|---|
| Acid fuchsin | 1 | g |
| Acetic acid | 1 | mL |
| Distilled water | 100 | mL |
Stock solution 2
| Material | Amount | |
|---|---|---|
| Fast yellow | 3 | g |
| Acetic acid | 1 | mL |
| Distilled water | 100 | mL |
Stock solution 3
| Material | Amount | |
|---|---|---|
| Phosphomolybdic acid | 1 | g |
| Distilled water | 100 | mL |
Working solution A
| Material | Amount | |
|---|---|---|
| Stock solution 1 | 30 | mL |
| Stock solution 2 | 30 | mL |
| Stock solution 3 | 30 | mL |
Working solution B
| Material | Amount | |
|---|---|---|
| Acetic acid | 1 | mL |
| Distilled water | 100 | mL |
Working solution C
| Material | Amount | |
|---|---|---|
| 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.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with an acid resistant nuclear stain.
- Rinse well with tap water.
- Place into solution A for 5 minute.
- Rinse quickly with distilled water.
- Place into solution B for 5 minutes.
- With a pipette, drop solution C onto the slide for 30 seconds.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Elastic – pink
- Cytoplasm – red
- Collagen – yellow
- Nuclei – black
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- 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, v. 10, p. 404.
