Gadsdon's Alum Hematoxylin
This formula is by Prof. Derek Gadsdon at the Sheffield Children’s Hospital, UK.
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 5.5 g | Dye |
| Potassium alum | 60 g | Mordant |
| Distilled water | 610 mL | Solvent |
| 100% ethanol | 100 mL | Solvent |
| Glycerol | 300 mL | Stabiliser |
| Sodium iodate | 0.5 g | Oxidant |
| Glacial acetic acid | 20 mL | Acidifier |
Compounding procedures
- Dissolve the hematoxylin in 100 mL ethanol.
- Dissolve the Alum in 600 mL water.
- Dissolve the sodium iodate in 10 mL water.
- Combine the hematoxylin and Alum solutions. Mix well.
- Add the sodium iodate solution. Mix well.
- Add the acetic acid. Mix well.
- Leave overnight.
- Add the glycerol. Mix well.
- Filter before use.
- The solution may be used immediately.
- It is stable at room temperature for at least 6 months.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for the appropriate time.
- Rinse well with water.
- Differentiate with acid ethanol if using regressively.
- 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
- This solution may be used regressively or progressively.
- Regressively, staining time is 6 minutes.
- Progressively, staining time is 3 minutes.
- Frozen sections will stain in 1 minute.
- As with many strong formulations mucins may stain blue.
- Acid ethanol is 0.5% – 1% hydrochloric acid in 70% ethanol.
- 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.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Histopathology laboratory, Red Cross Children’s Hospital.
Gadsdon’s Haematoxylin Method
Histonet communication, Nov, 1996 - Jim Elsam, HTEQA Services.
Gadsdon’s Hx
Histonet communication, Nov, 1996
