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H&E Staining is intended for use in histology and cytology applications. Included in this staining is a newly formulated Eosin that provides the benefits of a traditional alcoholic formula with significant improvements in usability. Advantages include lower evaporation rate, better color patterns, reduced tendency to spill over container, hands, and countertops, and improved surface tension to remain on tissue section. The Hematoxylin produces crisp, intense blue nuclei providing optimal contrast to the Eosin stained cytoplasm.
1) Deparaffinize sections, 2 changes of xylene, 10 minutes each.
2) Re-hydrate in 2 changes of absolute alcohol, 5 minutes each.
3) 95% alcohol for 2 minutes and 70% alcohol for 2 minutes.
4) Wash briefly in distilled water.
5) Stain in Mayer hematoxylin solution for 8 minutes.
6) Wash in warm running tap water for 10 minutes.
7) Rinse in distilled water.
8) Rinse in 95% alcohol, 10 dips.
9) Counterstain in eosin-phloxine B solution (or eosin Y solution) for 30 seconds to 1 minute.
10) Dehydrate through 95% alcohol, 2 changes of absolute alcohol, 5 minutes each.
11) Clear in 2 changes of xylene, 5 minutes each.
12) Mount with xylene based mounting medium.
1) Nuclei: blue
2) Cytoplasm: pink to red
Leica ST5020 Autostainer w/ CV5030 Slide Coverslipper & TS5025 Transfer Station