Heat the water. Then add in order:-
Hematoxylin, sodium iodate, alum, citric acid and chloral hydrate.
Dissolve each ingredient before adding the next.
Cool to room temperature and filter.
It may be used immediately.
Bring sections to water with xylene and ethanol.
Place into the staining solution for 5-20 minutes.
Rinse with water and blue.
Rinse well with water.
Counterstain if desired.
Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Nuclei – blue
Background – as counterstain or unstained
Putt gives no information regarding staining time and characteristics,
but the similarity of the formula to Mayer's (Langeron's)
hemalum suggests it is progressive and selectively nuclear.
A staining time of 10-20 minutes should be adequate.
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.
F. A. Putt Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA