Lugol’s Alum Hematoxylin
Materials
Lugol’s Iodide
| Material | Amount | Function |
|---|---|---|
| Iodine | 2 g | Oxidant |
| Potassium iodide | 4 g | |
| Distilled water | 100 mL | Solvent |
Hematoxylin
| Material | Amount | Function |
|---|---|---|
| 95% ethanol | 100 mL | Solvent |
| Hematoxylin | 10 g | Dye |
Alum
| Material | Amount | Function |
|---|---|---|
| Potassium alum | 6 g | Mordant |
| Distilled water | 500 mL | Solvent |
Working solution
| Material | Amount |
|---|---|
| Hematoxylin | 1.5 mL |
| Lugol’s iodine | 1.5 mL |
| Alum | 100 mL |
Compounding Procedure
- Make each solution as listed.
- Combine as directed above.
- Leave for 15 minutes before use.
Protocol
- Bring sections to distilled water with xylene and ethanol.
- Place into the staining solution for 5-10 minutes.
- 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
- Read these comments on Lugol’s iodine.
- The solution is usable for a minimum of one week.
- The working solution will stain at least 100 slides.
- Since the solution is not acidified, a distilled water rinse before staining will reduce alkaline carry over.
- The authors named this solution Lugol’s Hematoxylin.
- 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
- Clarke, G. and Dodds, H. M., (1983)
Lugol’s Hematoxylin.
Stain Technology, v 58, Nº 4, p. 232
