Var I
Dissolve the dye into the ethanol, and the alum into the water.
Combine the solutions in an Erlenmeyer flask and slowly add the mercuric oxide
Reheat until the solution changes colour to a dark purple.
Remove from heat and cool rapidly by swirling the flask in cold water.
When cool, add the acetic acid.
Var II
Dissolve the alum into the water.
Add the hematoxylin, sodium iodate, citric acid and chloral hydrate in that order.
Filter before use.
Method
Bring sections to water with xylene and ethanol.
Place into the staining solution for the specified time.
Var I for 10 minutes.
Var II for 8-10 seconds.
Rinse well with water.
Differentiate Var I with acid ethanol for a few seconds.
Rinse with water and blue.
Rinse well with water.
Counterstain if desired.
Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected results
Nuclei – blue
Background – as counterstain or unstained
Notes
Var I is a modification of Harris' hemalum and recommended for double embedded tissues.
Var II is a modification of Mayer's hemalum., but the quadrupling of the
dye content may have changed the hemalum from a progressive
to a regressive formulation. The very short staining time of 8-10 seconds would support this.
It could likely be applied for some minutes, then lightly differentiated with acid ethanol as with other
regressive formulations.
The use of mercuric oxide as an oxidant is now deprecated due to its toxicity.
Var I could be modified by using sodium iodate at about 0.3 grams instead.
Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
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 Molnar, L. N., Modification of Harris hematoxylin for sections from tissue double embedded with nitrocellulose and paraffin.
Histologic, v 5, Nº 1, January, 1975
Molnar, L. N., Modification of Mayer's hematoxylin-eosin method.
Histologic, v 6, Nº 4, October, 1976
