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Proescher & Arkush Iron Alum-Celestine Blue for Nuclei

Proescher & Arkush Iron Alum-Celestine Blue

for Nuclei



  • An eosin solution, or other counterstain
  • Iron alum-celestine blue
    Ferric ammonium sulphate5g
    Celestine blue B0.5g
    Distilled water100mL

    Dissolve the ferric ammonium sulphate in the distilled water. Add the celestine blue B. Boil for 5 minutes. Cool and filter.

Tissue Sample

5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.


  1. Bring sections to water via xylene and ethanol.
  2. Place into the iron alum-celestine blue solution for 3 minutes to 2 hours.
  3. Wash with tap water.
  4. Counterstain with eosin or other counterstain.
  5. Dehydrate with 95% and absolute ethanols.
  6. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue
  • Cytoplasm  –  pink


  • The iron alum-celestine blue solution is stable for a few months.
  • A staining time of 5-10 minutes is usually satisfactory for formalin fixed tissues.
  • This method has been recommended as a substitute for Hematoxylin and Eosin.
  • Proescher & Arkush also recommended the dyes gallamine blue and gallocyanin. Neither dye is as stable as celestine blue, although the color with gallocyanin most closely resembles alum hematoxylin of all three dyes.

Safety Note

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


  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Proescher and Arkush, (1928)
    Stain Technology, v. 3, pp. 36