Hematoxylin and Eosin (H&E) staining is used most commonly in histology. Hematoxylin is a basic dye and stains basophilic structure. Eosin Y is a fluorescent red dye is used to stain cytoplasm, collagen and muscle fiber. Slides stained with hematoxylin and eosin can only be mounted with organic mounting mediums. H&E stains slides are not compatible with aqueous mounting medium.
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H and E staining: Follow the procedure as outlined for H and E staining. We use the following protocol:
- Deparafinize formalin-fixed paraffin-embedded (FFPE) tissue sections.
- Stain with hematoxylin for 4 minutes (2X), wash 4-5X with tap water, and rinse 2 X with de/di water.
- Dip in 10 % acetic acid 2X, wash with de/di water, wash with Immuno Automation buffer (ImmunoBioScience AR-6561), PBS or any buffer with pH 7.4-8
- Rinse 2X with 70% ethanol, apply Eosin stain 1-5 minutes, and wash with 70% ethanol to remove excess Eosin.
- Rinse 2X with 95% ethanol, 2X with 100% ethanol, xylene or any dehydrating agent.
- The slide can be mounted with organic mounting medium or our Organo /limonene (AR-6504) mounting medium.