Rozas' Iron Hematoxylin
Rozas’ iron hematoxylin is of interest as it contains aluminum and iron salts in one solution. Both are mordants used with hematoxylin, and there will presumably be some kind of competition for available binding sites which could modify the solution’s staining characteristics. Since iron can be used to displace aluminum as a mordant for hematoxylin, it is likely that the solution stains more as an iron hematoxylin than as an aluminum hematoxylin.
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 0.6 g | Dye |
| Ferric ammonium sulfate | 1 g | Mordant |
| Aluminum chloride | 1.2 g | Mordant |
| Distilled water | 74 mL | Solvent |
| 95% ethanol | 6 mL | Solvent |
| Glycerol | 20 mL | Stabiliser |
Solution B
| Material | Amount | Function |
|---|---|---|
| Ferric ammonium sulfate | 20 g | Mordant |
| Distilled water | 100 mL | Solvent |
Compounding Procedure
- For solution A, dissolve the hematoxylin in the ethanol.
- Dissolve the ferric ammonium sulfate and the aluminum chloride in the water.
- Combine, then add the glycerol.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into solution A for 12-24 hours.
- Place into solution B until differentiated.
- Wash well in running tap water to blue.
- Rinse with distilled water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Background – as counterstain or unstained
Notes
- The presence of glycerol indicates that solution A may be stable for some time.
- The staining procedure indicates that the technique is intended as a primary stain. It is likely a modification of Heidenhain’s method and would demonstrate much the same material.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
Citing:
Rozas, (1935)
Zietschrift für wissenschaftliche Mikroskopie und für mikroskopische Technik,
v.52, p.1. Leipzig.
