Régaud's Iron Hematoxylin
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Ferric ammonium sulfate | 5 g | Mordant |
| Distilled water | 100 mL | Solvent |
Solution B
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 1 g | Dye |
| 90% ethanol | 10 mL | Solvent |
| Distilled water | 80 mL | Solvent |
| Glycerol | 10 mL | Stabiliser |
Solution C
| Material | Amount | Function |
|---|---|---|
| Potassium ferricyanide | 2.5 g | Bleach |
| Sodium borate | 2 g | pH control |
| Distilled water | 100 mL | Solvent |
Compounding Procedure
- Make each solution separately.
- Solutions A and B may be reused provided they are not contaminated by each other.
- Solution C should not be reused.
Protocol
- Bring sections to distilled water with xylene and ethanol.
- Place into solution A at 50°C for 30 minutes.
- Rinse well with distilled water.
- Place into solution B at 50°C for 30 minutes.
- Rinse well with distilled water.
- Place into solution C until differentiated.
- Wash in running tap water until blue.
- Counterstain as required.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Background – as counterstain or unstained
Notes
- The solutions are stable for some time.
- Solution C is Weigert’s ferricyanide bleach and should be used fresh.
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.
