Hematoxylin, Immuno/Histo Aqueous

A. H and E staining: Follow the procedure as outlined for H and E staining. We use the following protocol:
1. Deparafinize formalin-fixed paraffin-embedded (FFPE) tissue sections.
2. Stain with hematoxylin for 4 minutes (2X), wash 4-5X with tap water, and rinse 2 X with deionized/distilled (de/di) water.
3. Dip in 10 % acetic acid 2X, wash with de/di water, wash with Immuno Automation buffer (ImmunoBioScience), PBS or any buffer with pH 7.4-8.
4. Rinse 2X with 70% ethanol, apply Eosin stain 1-5 minutes, and wash with 70% ethanol to remove excess Eosin.
5. Rinse 2X with 95% ethanol, 2X with 100% ethanol, xylene or any dehydrating agent.
6. The slide can be mounted with organic mounting medium or our Organo (limonene) mounting medium (AR-6504).
B. Immunohistochemistry:
1. After chromogen staining, wash slide 2-3X with de/di water, 2X with buffer and 2X with de/di.water
2. Apply Immuno hematoxylin for 30-60 seconds; wash with tap water 5-7X.
3. Soak the slides in Immuno Automation buffer (AR-6561), PBS or any buffer with pH 7.4-8 for 2-3 minutes. Hematoxylin changes color from purple red to blue.
4. Wash slide 2-3X with de/di water.
5. The slides can be mounted with aqueous mounting medium (ImmunoHistoMount or equivalent) for chromogens like AEC, DAB, Fast red, BCIP/INT, or Super Fast red.
6. DAB and Super Fast can be mounted with Organic mounting mediums like Organo mount (limonene) or equivalent after dehydrating the tissue with alcohols and organic solvents.