Smith's Vanadium Hematoxylin
When differentiated with dilute lithium carbonate, this solution demonstrates basic proteins, including nuclear histones.
Materials
Material | Amount | Function |
---|---|---|
Hematoxylin | 50 mg | Dye |
Ethanol, 100% | 5 mL | Solvent |
Glycerol | 10 mL | Solvent |
Distilled water | 35 mL | Solvent |
Ammonium metavanadate | 200 mg | Solvent |
Compounding procedure
- Dissolve the hematoxylin in the ethanol, then add the glycerol and water in sequence.
- Add the ammonium vanadate and stir for 30 minutes.
- The ammonium vanadate may not completely dissolve. The pH is 6.2.
Tissue Sample
Paraffin sections of Schaudinn, Bouin or formalin fixed tissues are suitable. Sections of Zenker or Helly fixed tissues must be soaked in saturated aqueous lithium carbonate for 4 hours to stain satisfactorily.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the vanadate hematoxylin for 30 minutes.
- Place directly into 0.08% aqueous lithium carbonate for the appropriate time.
Differentiation times in lithium carbonateFixative Time Bouin 2 minutes Formalin, 10% 2 minutes Helly 10 minutes Schaudinn 5 minutes Zenker 10 minutes - Briefly rinse with distilled water for 2 seconds.
- Optionally, counterstain if wished.
- Dehydrate with 70%, 95% and absolute ethanols.
- Clear with xylene and mount with a resinous medium (Permount specified).
Expected Results
- Nuclear histone – blue
- Ribosomal protein – paler blue
- Other protein – as counterstain or unstained
Notes
- Counterstaining can be accomplished at step 5 with 1% aqueous eosin Y, phloxine B or erythrosin B.
- Alternatively, metanil yellow at 0.1% in 95% ethanol containing 0.1% acetic acid may be used between the 70% and 95% ethanols in step 6.
- Counterstaining with 0.01% aqueous safranin O stained RNA intensely, giving good contrast between blue nuclear histone and endoplasmic reticulum.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Smith, A. A., (1995)
A vanadate hematoxylin stain for basic proteins.
Biotechnic and Histochemistry, v. 70, Nº 5, p. 5.