Maxwell's Trichrome
for Pituitary Cells
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Acid fuchsin | 1 | g |
| Distilled water | 100 | mL |
Solution B
| Material | Amount | |
|---|---|---|
| Ammonia (0.880) | 0.02 | mL |
| Distilled water | 100 | mL |
Solution C
| Material | Amount | |
|---|---|---|
| Hydrochloric acid (Conc.) | 0.1 | mL |
| Distilled water | 100 | mL |
Solution D
| Material | Amount | |
|---|---|---|
| Phosphomolybdic acid | 0.5 | g |
| Distilled water | 100 | mL |
Solution E
| Material | Amount | |
|---|---|---|
| Orange G | 2 | g |
| Aniline blue | 2 | g |
| Phosphomolybdic acid | 1 | g |
| Distilled water | 100 | mL |
Maxwell’s fluid
| Material | Amount | |
|---|---|---|
| Cedarwood oil | 30 | mL |
| Thyme oil | 40 | mL |
| Xylene | 15 | mL |
| Ethanol, absolute | 15 | mL |
Tissue Sample
Paraffin sections at 5µ are suitable. Heidenhain’s (Zenker formol) fixation was specified. Other mercuric chloride fixatives would likely be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into solution A for 30 minute.
- Rinse with distilled water.
- Place into solution B until differentiated.
- Place into solution C for a few seconds.
- Place into solution D for 3 minutes.
- Place into solution E for 60 minutes.
- Rinse with distilled water.
- Differentiate with 95% ethanol.
- Dehydrate with ethanol.
- Clear with Maxwell’s fluid or xylene.
- Mount with Canada balsam or other resinous medium.
Expected Results
- Nuclei – red
- Acidophils – orange red
- basophils – blue
Notes
- Maxwell’s fluid was originally specified for clearing, but this is probably not necessary.
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:
Maxwell, (1938)
Stain Technology, vol. 13, pp. 93
