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Hansen’s Iron Hematoxylin

Hansen's Iron Hematoxylin

8
steps
3
materials

Materials

MaterialAmountFunction
Hematoxylin0.75 gDye
Ferric ammonium sulfate4.5 gMordant
Distilled water100 mLSolvent

Compounding Procedure

  1. Dissolve the iron alum in 65 mL of the water.
  2. Dissolve the hematoxylin in the remaining water.
  3. Combine the solutions and bring to a boil.
  4. Cool and filter.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for an appropriate time.
  3. Rinse with tap water.
  4. Treat briefly in 1% hydrochloric acid in 70% ethanol.
  5. Wash well in running tap water to blue.
  6. Rinse with distilled water.
  7. Counterstain if desired.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  black
  • Background  –  as counterstain or unstained

Notes

  • The stability of the solution is not given, but the simplicity of the formula indicates it would not be stable for too long.
  • Although a staining time was not given, 30 minutes should be sufficient. Determine by trial.

Safety Note

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

References

  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Citing:
    Hansen, (1905)
    Zeitschrift für wissenschaftliche Mikroskopie und für mikroskopische Technik,
    v.22, p.55. Leipzig.