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Hamilton Alum Hematoxylin

Hamilton Alum Hematoxylin

6
steps
6
materials

This formula is based on Langeron’s 1942 (Mayer’s) formula. It has been referred to as Hamilton Alum hematoxylin as it was received via the internet from a histotechnologist at the Hamilton Health Sciences Corporation, Hamilton, Ontario, Canada.

Materials

MaterialAmountFunction
Hematoxylin2 gDye
Ammonium alum75 gMordant
Distilled water950 mLSolvent
100% ethanol50 mLSolvent
Sodium iodate0.3 gOxidant
Glacial acetic acid30 mLAcidifier

Compounding Procedure

  1. Dissolve the Alum in the water.
  2. Add the ethanol and mix well.
  3. Add the hematoxylin and dissolve.
  4. Add the sodium iodate and mix well.
  5. Leave overnight in the dark to ripen.
  6. Mix well, and add the acetic acid.
  7. Filter.
  8. The solution may be used immediately.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for 3 minutes.
  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

  • Potassium Alum may be used instead of ammonium alum.
  • The solution is progressive.
  • 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

Personal internet communication, source unidentified.