Mallory's Trichrome
for Muscle and Collagen
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Acid fuchsin | 1 | g |
| Distilled water | 100 | mL |
Solution B
| Material | Amount | |
|---|---|---|
| Phosphomolybdic acid | 1 | g |
| Distilled water | 100 | mL |
Solution C
| Material | Amount | |
|---|---|---|
| Orange G | 2 | g |
| Methyl blue | 0.5 | g |
| Oxalic acid | 2 | g |
| 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.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into solution A for 2 minute.
- Rinse with distilled water.
- Place into solution B for 2 minutes.
- Rinse quickly with distilled water.
- Place into solution C for 15 minutes.
- Wash well with distilled water.
- Dehydrate and differentiate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – red
- Erythrocytes – orange
- Muscle – red
- Collagen – blue
Notes
- Phosphotungstic acid may be substituted for phosphomolybdic acid in solution B.
- This is the method that introduced the concept, if not the name, of trichrome staining.
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:
Mallory, (1901)
Journal of Experimental Medicine, v.5, pp.15
And:
Mallory, (1936)
Stain technology, v.11, pp.101
