Rawitz'
Alum Hematoxylin Variants

Ingredient	1895a	1895b	1909	Function
Hematoxylin	10 g	–	–	Dye
Hematein	–	2.5 g	2 g	Dye
Potassium alum	10 g	–	–	Mordant
Ammonium alum	–	15 g	–	Mordant
Aluminum nitrate	–	–	20 g	Mordant
Distilled water	650 mL	500 mL	500 mL	Solvent
Glycerol	350 mL	500 mL	500 mL	Stabiliser

 

Compounding procedure
Dissolve the aluminum salt and dye in the water.
When dissolved, add glycerol.
The 1895a formula should be allowed to ripen.
The 1895b and 1909 formulas may be used immediately.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for an appropriate time.
3. Rinse well with water.
4. Differentiate with acid ethanol if necessary.
5. Rinse with water and blue.
6. Rinse well with water.
7. Counterstain if desired.
8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Background  –  as counterstain or unstained

Notes

1. The 1895b and 1909 formulae use hematein instead of hematoxylin.
2. The Microtomists Formulary and Guide gives the 1895b and 1909 formulae as containing 500 millilitres of glycerol. The Microtomists Vade-Mecum gives these two formulae as containing 500 grams glycerol. This equates to 400 millilitres based on a specific gravity of 1.25.
3. The staining time should be determined by trial.
4. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
5. Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.

 

Reference
Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.

Bolles Lee, A.. Edited by Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist's Vade-Mecum. 11 ed.,
Churchill, London, UK.

 

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