Bensley's Copper Hematoxylin
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Hematoxylin, | 1 g | Dye |
| Distilled water | 100 mL | Solvent |
Solution B
| Material | Amount | Function |
|---|---|---|
| Cupric acetate, | to saturation | Mordant |
| Distilled water | 100 mL | Solvent |
Solution C
| Material | Amount | Function |
|---|---|---|
| Potassium chromate, | 5 g | – |
| Distilled water | 100 mL | Solvent |
Solution D
| Material | Amount | Function |
|---|---|---|
| Weigert’s borax-ferricyanide | 20 mL | Differentiator |
| Distilled water | 80 mL | Diluent |
Compounding procedure
Make each solution separately.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into solution A for 5 minutes.
- Rinse with water.
- Dip into solution B.
- Rinse with water.
- Place into solution C for 2 minutes.
- Return to solution A for 1 minute.
- Place in solution D until differentiated.
- Rinse well with water.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei and other structures – blue-black
Notes
- After step 7, the sections should be intense black. Repeat steps 5-7 if necessary.
- Bensley recommended prestaining with Mayers mucicarmine.
- Best results are obtained with freshly made solutions
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Bensley R. R. and Bensley, S. H., (1938)
Handbook of Histological and Cytological Technique.
U. Chicago Press, Chicago, USA
