Delafield's Alum Hematoxylin
Materials
Material | Amount | Function |
---|---|---|
Hematoxylin | 6 g | Dye |
Ammonium alum | 90 g | Mordant |
Distilled water | 600 mL | Solvent |
95% ethanol | 200 mL | Solvent |
Glycerol | 150 mL | Stabiliser |
Compounding procedures
- Dissolve the hematoxylin in 50 mL ethanol.
- Dissolve the alum in the water.
- Add the two solutions.
- Leave, loosely stoppered, in a warm, sunlit place for one week.
- Filter, then add the glycerol and the rest of the ethanol.
- Leave, loosely stoppered, in a warm, sunlit place for 3 months.
- Filter and store in a tightly stoppered container in the dark.
- This solution is stable for years.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 20 minutes.
- Rinse well with water.
- Differentiate with acid ethanol if necessary.
- 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 solution may be chemically ripened by adding 0.5 g sodium iodate, but such solutions are considered to be inferior in longevity.
- Like many strong regressive formulations, cartilage, cement lines and mucins may be 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
- Culling, C.F.A., Allison, R.T. and Barr, W.T.
Cellular Pathology Technique, Ed.4.
Butterworth, London, UK. - Bancroft, J.D. and Stevens A. (1982)
Theory and practice of histological techniques Ed. 2
Churchill Livingstone, Edinburgh & London, UK.