Langeron's
Alum Hematoxylin Variants

Both of Langeron's formulae are modifications of Mayer's formulae. Langeron's 1924 formula is a modification of Mayer's 1896 formula, and Langeron's 1942 formula is a modification of Mayer's 1901 formula.

Langeron's 1942 formula is the solution usually meant when Mayer's hemalum is specified.

Ingredient	1924	1942	Function
Hematoxylin	4 g	1 g	Dye
Ammonium alum	50 g	–	Mordant
Potassium alum	–	50 g	Mordant
Distilled water	700 mL	1 L	Solvent
Glycerol	300 mL	–	Stabiliser
Sodium iodate	–	0.2 g	Oxidant
Glacial acetic acid	20 mL	–	Acidifier
Citric acid	–	1 g	Acidifier
Chloral hydrate	–	50 g	Acidifier

 

Compounding procedure

1924
Grind the hematoxylin with glycerol in a pestle and mortar.
Dissolve the other ingredients in water.
Add the hematoxylin paste.
Wash out the hematoxylin paste with the solution.
Leave months to ripen.

1942
Dissolve the alum and dye in the water.
Add the other ingredients.
Bring to the boil.
Cool to room temperature.
The solution may be used immediately.
Ammonium alum may be substituted for potassium alum.

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 the 1924 solution 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 1924 solution is regressive, and staining time should be determined by trial.
2. The 1942 solution is progressive. Staining time is 5-10 minutes.
3. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
4. 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.