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Delafield’s Alum Hematoxylin

Delafield's Alum Hematoxylin

8
steps
5
materials

Materials

MaterialAmountFunction
Hematoxylin6 gDye
Ammonium alum90 gMordant
Distilled water600 mLSolvent
95% ethanol200 mLSolvent
Glycerol150 mLStabiliser

Compounding procedures

  1. Dissolve the hematoxylin in 50 mL ethanol.
  2. Dissolve the alum in the water.
  3. Add the two solutions.
  4. Leave, loosely stoppered, in a warm, sunlit place for one week.
  5. Filter, then add the glycerol and the rest of the ethanol.
  6. Leave, loosely stoppered, in a warm, sunlit place for 3 months.
  7. Filter and store in a tightly stoppered container in the dark.
  8. This solution is stable for years.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for 20 minutes.
  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 solution may be chemically ripened by adding 0.5 g sodium iodate, but such solutions are considered to be inferior in longevity.
  • Like many strong regressive formulations, cartilage, cement lines and mucins may be blue.
  • 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. Culling, C.F.A., Allison, R.T. and Barr, W.T.
    Cellular Pathology Technique, Ed.4.
    Butterworth, London, UK.
  2. Bancroft, J.D. and Stevens A. (1982)
    Theory and practice of histological techniques Ed. 2
    Churchill Livingstone, Edinburgh & London, UK.