Compounding procedure
Dissolve the alum in the water.
Add the hematoxylin.
The solution is likely progressive, although this is not stated to be so.
Method
Bring sections to water with xylene and ethanol.
Place into the staining solution for a few 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.
Expected results
Nuclei – blue
Background – as counterstain or unstained
Notes
This solution is from the late 1800's and is now obsolete,
although the modern formula should stain satisfactorily.
Concentrated alcoholic hematoxylin would most likely have been
approximately 10% hematoxylin in absolute or 95% ethanol. This was used as it
afforded some degree of consistency in making solutions. It was made by soaking
logwood chips in ethanol until no more dye would dissolve out. Depending on the
sample of logwood and the amount of dye it contained, more than one batch may
have been necessary to saturate the ethanol. The solution was then ripened naturally.
The type of alum was not specified, the most likely being either
potassium or ammonium.
The formula calls for adding a "little alum" to 800 mL water.
Potassium alum saturates at about 14% in water, so 800 mL would contain about 112 g.
I have taken just less than 25% of the maximum (i.e. 25 grams) as being a "little".
Of course, it could be any amount between 1 and 112 grams.
The appropriate time should be determined by trial.
The instructions are to use full strength for a few minutes.
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 Arthur Bolles-Lee, (1885) The Microtomist's Vade-Mecum Originally published by: J & A Churchill, London, England. Republished by: Science Heritage Ltd., Lincolnwood, Illinois, USA.
Susan Budavari, Editor, (1996) The Merck Index, Ed. 12
Merck & Co., Inc., Whitehouse Station, NJ, USA
