Masson's HES
General Oversight Stain
This is also known as the Hematoxylin Erythrosine Saffron stain.
Materials
- Regressive hemalum
- Solution A
Material Amount Erythrosine B 1 g Tap water 100 mL Dissolve the erythrosine B and filter. Preserve with a few drops drops of chloroform.
- Solution B
Material Amount Saffron 2 g Distilled water 100 mL Strong formalin 1 mL Tannic acid, 5% aqueous 1 mL 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.
Protocol
- Bring sections to water with xylene and ethanol.
- Treat with Bouin’s fluid if desired.
- Stain nuclei with hemalum, differentiate and blue.
- Wash well with water.
- Place in solution A for 2-5 minutes.
- Wash rapidly with tap water.
- Differentiate erythrosine with 70% ethanol for a few seconds to decolorize collagen.
- Wash rapidly with tap water.
- Place into solution B for 5 minutes.
- Wash rapidly with tap water.
- Dehydrate rapidly with absolute ethanol.
- 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
Notes
- 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.
References
- Biological Staining Methods, 6th ed. (1957)
Gurr, G. T.,
George T. Gurr, London, UK - Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
