Bosma's
Alum Hematoxylin

Ingredient Amount Function
Hematoxylin, 10% alc. 25 mL Dye
Ammonium alum 25 g Mordant
Tap water 865 mL Solvent
Diethylene glycol 100 mL Stabiliser
Sodium iodate 0.2 g Oxidant
Glacial acetic acid 10 mL Acidifier

 

Compounding procedure
Place 815 mL hot tap water in a large flask.
Add the ammonium alum, mix well to dissolve and cool to room temperature.
Add the alcoholic hematoxylin solution (ripened).
Add the sodium iodate dissolved in 50 mL cold tap water and mix well.
Add the diethylene glycol and mix well.
Add the acetic acid and mix well.
The pH should be 3.1 to 3.3 and the solution may be used immediately.

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 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

Notes

  1. The staining time should be determined by trial.
  2. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
  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
Bosma, Rob
A useful hematoxylin without toxic chemicals.
Histologic, V 18, Nº 1, January 1988

 


 

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