Protocol
Eosin
for Eosinophils
Materials
- Alum hematoxylin, such as Mayer’s
- Eosin Y ws, 1% in tap water
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei lightly with alum hematoxylin.
- Differentiate if necessary and blue.
- Place in eosin solution for 5 minutes.
- Wash well with tap water until sections are almost unstained.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Eosinophils – bright pink
- Background – pale pink to colorless
Notes
- The tap water should be hard and very slightly alkaline.
- Areas which have acidic or soft tap water should use Scott’s tap water substitute. If this is too alkaline, it may be diluted.
- The water wash is to remove eosin from connective tissues so that the eosinophil granules are the most prominent feature. A light pink background helps with orientation.
- Adjust staining and washing times to obtain suitable results.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
