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.

Solution A	Amount	Function
Hematoxylin	0.6 g	Dye
Ferric ammonium sulphate	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	Amount	Function
Ferric ammonium sulphate	20 g	Mordant
Distilled water	100 mL	Solvent

 

Compounding procedure
For solution A, dissolve the hematoxylin in the ethanol.
Dissolve the ferric ammonium sulphate and the aluminum chloride in the water.
Combine, then add the glycerol.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into solution A for 12-24 hours.
3. Place into solution B until differentiated.
4. Wash well in running tap water to blue.
5. Rinse with distilled water.
6. Counterstain if desired.
7. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  black
• Background  –  as counterstain or unstained

Notes

1. The presence of glycerol indicates that solution A may be stable for some time.
2. 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.

 

Reference

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.

 

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