Gill's Aluminum Hematoxylin Variants
Materials
| Material | Solution Strength | Function | ||
|---|---|---|---|---|
| Single (I) | Double (II) | Triple (III) | ||
| Hematoxylin | 2 g | 4 g | 6 g | Dye |
| Aluminum sulphate | 17.6 g | 70.4 g | 158.4 g | Mordant |
| Distilled water | 750 mL | 750 mL | 750 mL | Solvent |
| Ethylene glycol | 250 mL | 250 mL | 250 mL | Solvent |
| Sodium iodate | 0.2 g | 0.4 g | 0.6 g | Oxidant |
| Glacial acetic acid | 20 mL | 20 mL | 20 mL | Acidifier |
Compounding procedures
- Mix the ethylene glycol and water.
- Add the hematoxylin, then the sodium iodate.
- Add the aluminum sulphate and the acetic acid
- Stir for an hour at room temperature.
- Filter before use.
- The solutions may be used immediately, and are stable for approximately one year.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for an appropriate time.
- Rinse with water and blue.
- Rinse well with water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Background – as counterstain or unstained
Notes
- The three formulations are for progressive staining.
- The single strength solution is for cytology. Staining time is 2 minutes. The double strength solution is for paraffin sections. Staining time is 3 minutes. The triple strength solution is for paraffin sections. Staining time is 1.5 minutes.
- Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.
- The aluminum sulphate should be Al2(SO4)3.18 H2O. Adjust the amount if different.
- Similarly, the hematoxylin should be anhydrous. If hydrated (C16H14O6.3 H2O) use 2.36 g, 4.72 g and 7.08 g respectively.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling, C.F.A., Allison, R.T. and Barr, W.T.
Cellular Pathology Technique, Ed.4.
Butterworth, London, UK.
