Bensley's trichrome
for Muscle and Collagen
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Acid fuchsin | To saturation | |
| Aniline water | 100 | mL |
Solution Preparation
- Add about 20 g dye to the aniline water.
- Shake intermittently for 48 hours. Filter.
Solution B
| Material | Amount | |
|---|---|---|
| Phosphomolybdic acid | 1 | g |
| Distilled water | 100 | mL |
Solution C
| Material | Amount | |
|---|---|---|
| Orange G | 2 | g |
| Aniline blue | 0.5 | 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 10 minute.
- Rinse with distilled water.
- Place into solution B for 10 minutes.
- Rinse quickly with distilled water.
- Place into solution C for 60 minutes.
- Rinse with 95% ethanol until clouds of dye cease being extracted.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – red
- Erythrocytes – orange
- Cytoplasm – red
- Collagen – blue
Notes
- Solution A is Altmann’s acid fuchsin, originally formulated for staining mitochondria.
- This method does not specify an acid resistant nuclear stain. Doing so before staining with solution A may improve the technique.
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, F. B., (1938),
Pathological technique.
Saunders, Philadelphia, Pa, USA
