Masson's HES
General Oversight stain

Also known as the Hematoxylin Erythrosine Saffron stain.

Solutions

Regressive hemalum

Solution A
Erythrosine B	1	g
Tap water	100	mL

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

Solution B
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. 5µ 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 discolouration.

Method

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

Notes

1. 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 colouring. Usually, whole stigmata are more effective than ground saffron.
2. Eosin B may be substituted for erythrosine B.

 

Reference
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.

Translate in Google Translate Instructions