Gill's
Aluminum Hematoxylin Variants

Ingredient	Single (I)	Double (II)	Triple (III)	Function
Hematoxylin	2 g	4 g	6 g	Dye
Aluminum sulphate	17.6 g	70.4 g	158.4 g	Mordant
Distilled water	750 mL	750 mL	750 mL	Solvent
Ethylene glycol	250 mL	250 mL	250 mL	Solvent
Sodium iodate	0.2 g	0.4 g	0.6 g	Oxidant
Glacial acetic acid	20 mL	20 mL	20 mL	Acidifier

 

Compounding procedure
Mix the ethylene glycol and water.
Add the hematoxylin, then the sodium iodate.
Add the aluminum sulphate and the acetic acid
Stir for an hour at room temperature.
Filter before use.
The solutions may be used immediately, and are stable for approximately one year.

Method

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

Expected results

• Nuclei  –  blue
• Background  –  as counterstain or unstained

Notes

1. The three formulations are for progressive staining.
2. The single strength solution is for cytology. Staining time is 2 minutes.
   The double strength solution is for paraffin sections. Staining time is 3 minutes.
   The triple strength solution is for paraffin sections. Staining time is 1.5 minutes.
3. 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.
4. The aluminum sulphate should be Al₂(SO₄)₃.18 H₂O. Adjust the amount if different.
5. Similarly, the hematoxylin should be anhydrous.
   If hydrated (C₁₆H₁₄O₆.3 H₂O) use 2.36 g, 4.72 g and 7.08 g respectively.

Reference
Culling, C.F.A., Allison, R.T. and Barr, W.T.
Cellular Pathology Technique, Ed.4.
Butterworth, London, UK.

 

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