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 (ImmunoBioScience 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.

This uniquely formulated immuno/histo hematoxylin is an excellent counterstain for immunohistochemistry (IHC) and histochemistry staining of nuclei. It is an aqueous solution and does not contain any alcohol or mercury compounds. The bright blue color is stable for 2-3 years if kept in the dark. The counterstain can be used with HRP chromogens, AEC, DAB and alkaline phosphatase chromogens, fast red, BCIP/INT and super fast red (ImmunoBioScience). This hematoxylin is also suitable for H and E staining in histology. Sides stained with this hematoxylin can be mounted with aqueous or organic mounting mediums.