McLachlan's
Alum Hematoxylin

This formulation has been named in honor of Dr. H.K.I. McLachlan FRCPA at the request of the originator, Mike Rentsch.

Ingredient	Var I	Var II	Function
Hematoxylin	2 g	2 g	Dye
Aluminum sulphate	17.5 g	–	Mordant
Ammonium alum	–	25 g	Mordant
Distilled water	700 mL	700 mL	Solvent
Glycerol	300 mL	300 mL	Stabiliser
Sodium iodate	0.2 g	0.2 g	Oxidant
Glacial acetic acid	20 mL	20 mL	Acidifier

 

Compounding procedure
Dissolve the aluminum sulphate or the alum in about 500 mL water.
Add the hematoxylin and dissolve.
Add the acetic acid and mix well.
Warm the glycerol to reduce viscosity, and add to the mixture.
Mix well, then make up to 1 litre with water.
Add the iodate and mix for 5-10 minutes.
Store in a tightly capped bottle in the dark.
The solution remains usable for 2-5 years.

Method

1. Bring sections to water with xylene and ethanol.
2. Place paraffin sections in the staining solution for 2-4 minutes
   and frozen sections for 40-60 seconds.
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. This is a progressive solution.
2. 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
Rentsch, M.
Australian Biostain P/L.
Personal communication via the internet.