ZILLION: A novel zebrafish model for studying inflammation and infection in Inflammatory Bowel Disease

$393
Raised of $5,750 Goal
7%
Ended on 6/03/18
Campaign Ended
  • $393
    pledged
  • 7%
    funded
  • Finished
    on 6/03/18

Automated Hematoxylin and Eosin (H&E) Staining

An Autostainer XL (ST5010, Leica Biosystems, Nussloch, Germany) was used to perform standard H&E staining. The automated staining protocol consisted of the following steps: three changes of xylenes (2, 2 and 1 minutes), rehydration in an ethanol/water gradient (100%, 100%, 95% ethanol for 2, 1 minutes and 20 seconds, respectively), followed by washing with deionized (DI) water (2 minutes). The rehydrated slides were stained with Gill 3 Hematoxylin (12 minutes; Sigma-Aldrich), washed with DI water (2 minutes), followed by 0.17% Acid Alcohol (1 minutes), washed with DI water (1 minutes), Ammonia water (20 seconds), washed with DI water (1 min), and 95% ethanol (10 seconds). Staining with Eosin (45 seconds; Sigma-Aldrich) was completed, followed by four washes with 100% ethanol (1 minute each), and four washes with xylene (1 minute each). Lastly, H&E stained slides were cover slipped with Cytoseal XYL (Thermo Fisher Scientific) and allowed to dry flat prior to imaging.


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