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Bosma’s Alum Hematoxylin

Bosma's Alum Hematoxylin

8
steps
6
materials

Materials

MaterialAmountFunction
Hematoxylin, 10% alc.25 mLDye
Ammonium alum25 gMordant
Tap water865 mLSolvent
Diethylene glycol100 mLStabiliser
Sodium iodate0.2 gOxidant
Glacial acetic acid10 mLAcidifier

Compounding procedure

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

Protocol

  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

  • Nuclei  –  blue
  • Background  –  as counterstain or unstained

Notes

  • The staining time should be determined by trial.
  • 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.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.

References

  1. Bosma, Rob
    A useful hematoxylin without toxic chemicals.
    Histologic, V 18, Nº 1, January 1988