McNulty-Smith's Zirconyl Hematoxylin
This hematoxylin substitutes for alcian blue pH 2.5 for staining acid mucins.
Materials
| Ingredient | Amount | Function |
|---|---|---|
| Hematoxylin | 100 mg | Dye |
| Ethanol, absolute | 5 mL | Solvent |
| Sodium iodate, 0.5% aqu. | 5 mL | Oxidant |
| Zirconyl chloride, octahydrate | 400 mg | Mordant |
| Distilled Water | 22.5 mL | Solvent |
| Glycerol | 7.5 mL | Anti-oxidant |
Compounding Procedure
- Dissolve the hematoxylin in the ethanol.
- Add the sodium iodate solution (freshly made).
- Add the zirconyl chloride.
- Combine the glycerol and water, and add to the solution.
- Stir for 5 minutes.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 10 minutes.
- Wash with distilled water, two changes of 2 minutes each.
- Counterstain in methylene green for 5 minutes.
- Wash with distilled water, two changes of 2 minutes each.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – green
- Acid mucins – Purple
Notes
- Formalin fixed, paraffin sections are suitable.
- The counterstain recommended was 0.05% methylene green in 2.4% boric acid.
- This method was recommended as a substitute for alcian blue staining of acid mucins (pH 2.5).
- Staining may be removed with 1% hydrochloric acid in 70% ethanol.
- The solution above is McNulty’s modification of Smith’s original, and stains more intensely:
- Hematoxylin, 100 mg
- Absolute ethanol, 5 mL
- 0.1% aqueous sodium iodate, 5 mL
- Zirconyl chloride octahydrate, 400 mg
- 22% aqueous glycerol, 45 mL
- Glacial acetic acid, 5 mL
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Smith, A. A., (1999)
Zirconyl hematoxylin staining of acidic mucins.
Journal of Histochemistry and Cytochemistry, v. 47, p. 1645. - McNulty, J. M. and Smith, A. A., (2004)
An improved formulation of the zirconyl hematoxylin stain for acidic mucins.
Biotechnic and Histochemistry, v. 79, Nº 5, p. 191. - McNulty, J. M., Kambour, M. J. and Smith, A. A., (2004)
Use of an improved zirconyl hematoxylin stain in the diagnosis of Barrett’s esophagus.
J. Cell. Mol. Med., v. 8, Nº 3, p. 382.
