Bencosme's
Alum Hematein

Original
Ingredient	Amount	Function
Hematein	2 g	Dye
Potassium alum	133 g	Mordant
Distilled water	920 mL	Solvent
Glacial acetic acid	20 mL	Acidifier

Variant
Ingredient	Amount	Function
Hematein	2.5 g	Dye
Potassium alum	120 g	Mordant
Distilled water	1 L	Solvent
Glacial acetic acid	10 mL	Acidifier

Compounding procedure
Add the alum to the water in a 2 L flask and bring to the boil for 1 minute.
Add the hematein and mix by swirling.
Place an inverted funnel over the flask and simmer for 10 minutes, shaking frequently.
Cool to room temperature and add the acetic acid.
Filter before use and twice weekly.
It may be used immediately.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for 2 minutes.
3. Rinse with water.
4. Differentiate with 0.5% hydrochloric acid in 70% ethanol for one second.
5. Rinse well with water and blue.
6. Rinse well with water.
7. Counterstain with HPS.
8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Background  –  shades of red and yellow.

Notes

1. Note that this solution uses hematein rather than hematoxylin. Hematoxylin could be used, but would require 0.4 grams sodium iodate to be added along with the dye for complete oxidation in the original solution, although 0.3 grams might be more appropriate to extend the life. Comparable amounts would be 0.5 g and 0.4 g respectively for the variant formula.
2. When freshly made this solution stains in 2 minutes. Staining time slowly increases to 10 minutes as the solution ages. Life is about a month.
3. 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
Medical Laboratory Technology, 2nd ed. (1969)
Lynch M.J., Raphael S.S., Mellor L.D., Spare P.D. and Inwood M.J.H.,
W. B. Saunders Co., Toronto, On., Canada

 

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