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Masson’s HES General Oversight Stain

Masson's HES

General Oversight Stain


This is also known as the Hematoxylin Erythrosine Saffron stain.


  • Regressive hemalum
  • Solution A
    Erythrosine B1g
    Tap water100mL

    Dissolve the erythrosine B and filter. Preserve with a few drops drops of chloroform.

  • Solution B
    Distilled water100mL
    Strong formalin1mL
    Tannic acid, 5% aqueous1mL

    Add the saffron stigmata to the water and heat in a boiling water bath for one hour. Filter, and add the formalin and tannic acid. Life is a few weeks.

Tissue Sample

No particular fixative was specified. 5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable but results may be improved by refixing in Bouin’s fixative or saturated aqueous picric acid at 56°C for one hour prior to staining, then washing the sections in tap water to remove all yellow discoloration.


  1. Bring sections to water with xylene and ethanol.
  2. Treat with Bouin’s fluid if desired.
  3. Stain nuclei with hemalum, differentiate and blue.
  4. Wash well with water.
  5. Place in solution A for 2-5 minutes.
  6. Wash rapidly with tap water.
  7. Differentiate erythrosine with 70% ethanol for a few seconds to decolorize collagen.
  8. Wash rapidly with tap water.
  9. Place into solution B for 5 minutes.
  10. Wash rapidly with tap water.
  11. Dehydrate rapidly with absolute ethanol.
  12. Clear with xylene and mount with a synthetic resinous medium.

Expected Results

  • Nuclei  –  blue
  • Cytoplasm  –  red shades
  • Muscle  –  pink
  • Elastic fibres  –  pink
  • Collagen  –  orange-yellow


  • Saffron is expensive. It may be available in East Indian grocery stores or health food stores as its most common use today is as a spice and food coloring. Usually, whole stigmata are more effective than ground saffron.
  • Eosin B or phloxine B may be substituted for erythrosine B.

Safety Note

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


  1. Biological Staining Methods, 6th ed. (1957)
    Gurr, G. T.,
    George T. Gurr, London, UK
  2. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.