Gabe's Aldehyde Fuchsin
Materials
Stock powder
| Material | Amount | |
|---|---|---|
| Pararosanilin | 5 | g |
| Paraldehyde | 5 | mL |
| Hydrochloric acid | 10 | mL |
| Distilled water | 1 | L |
Dye Preparation Procedures
- Add the dye to the water and boil.
- Cool to room temperature.
- Add paraldehyde and hydrochloric acid.
- Ripen at RT two days. Filter. Wash precipitate with 50mL distilled water. Dry the ppt and paper at 60°C.
- Store in a labelled container. Stable for about ten years.
Working solution
| Material | Amount | |
|---|---|---|
| Stock powder | 0.25 | g |
| Ethanol, 70% | 200 | mL |
| Acetic acid, glacial | 2 | mL |
Acid ethanol
| Material | Amount | |
|---|---|---|
| Hydrochloric acid, conc. | 1 | mL |
| Ethanol, 95% | 200 | mL |
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place in the working solution for 5 minutes.
- Rinse well with running tap water.
- Rinse with acid ethanol for 20-30 seconds to remove excess dye.
- Counterstain if wished.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Elastic fibres – purple
- Mast cells – purple
- Pituitary βcells – purple
- Sulphated mucins – purple
- Background – as the counterstain
- Nuclei – as the nuclear stain
Notes
- The basic fuchsin used for this solution should be one that is suitable for Schiff’s reagent, i.e., it should have a high pararosanilin content. Both methods involve forming a compound between an aldehyde and dye.
- Light counterstaining with a progressive alum hematoxylin and eosin is also suitable.
- Many other counterstains can be used, including methods such as Masson’s trichrome.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Kiernan. J.A., (1999)
Histological and histochemical methods: Theory and practice, Ed. 3
Butterworth Heinemann, Oxford, UK.
Citing:
Gabe, M., (1976)
Histological techniques.
Masson, Paris.
