Krutsay's Alum & Iron Hematoxylin
Krutsay’s Alum hematoxylin gives very highly selective staining of nuclei. It may also be easily converted into an iron hematoxylin for use as an acid resistant nuclear stain.
Materials
Alum hematoxylin
Material | Amount | Function |
---|---|---|
Hematoxylin | 1 g | Dye |
Potassium alum | 50 g | Mordant |
Distilled water | 1 L | Solvent |
Hydrochloric acid | 5 mL | Acidifier |
Sodium iodate | 0.2 g | Oxidant |
Iron conversion
Material | Amount | Function |
---|---|---|
Iron alum | 2 g | Mordant |
Hydrochloric acid | 0.5 mL | Acidifer |
Distilled water | 100 mL | Solvent |
Compounding Procedure
- Mix the reagents under Alum hematoxylin together.
- Bring to a boil
- Cool and filter.
- The solution may be used immediately.
Conversion to Iron Hematoxylin
- Mix the reagents under Iron conversion together, and dissolve.
- For use mix the following:
- Alum hematoxylin solution – 25 mL
- Iron conversion solution – 2 mL
Protocol
- Bring sections to water with xylene and ethanol.
- Place into either staining solution for 5 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 (alum) or black (iron)
- Background – as counterstain or unstained
Notes
- This is a progressive solution and is highly selective for nuclei.
- The iron Alum is (FeNH4(SO4)2.12 H2O)
- 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
- Humason, G. L.
Animal Tissue Techniques.
W. H. Freeman and Co., San Francisco, CA, USA