Skip to main content

Papamiltiades’ Alum Hematoxylin

Papamiltiades' Alum Hematoxylin

6
steps
7
materials

Materials

MaterialAmountFunction
Hematoxylin4 gDye
Aluminum sulphate10 gMordant
Distilled water900 mLSolvent
Zinc sulphate5 g
Potassium iodide4 gStabiliser
Glacial acetic acid32 mLAcidifier
Glycerol100 mLStabiliser

Compounding Procedure

  1. Dissolve the hematoxylin in 400 mL water.
  2. Dissolve the aluminum sulphate in 200 mL water.
  3. Dissolve the zinc sulphate in 100 mL water.
  4. Dissolve the potassium iodide in 100 mL water.
  5. Combine the four solutions, then add the acetic acid and glycerol.
  6. The solution may be used immediately, and is stable for about two months.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for an appropriate time.
  3. Rinse with water and blue.
  4. Rinse well with water.
  5. Counterstain if desired.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue
  • Background  –  as counterstain or unstained

Notes

  • The solution is progressive.
  • The purpose of the zinc sulphate and potassium iodide are not clear.
  • The staining time should be determined by trial.
  • 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. Putt, F.A.
    Manual of Histopathological Staining Methods
    John Wiley & Sons, New York, NY., USA