The screening workflow and automated image analysis to detect zebrafish embryos, and count haematopoetic stem cells in the tails. (A) Schematic of screening platform workflow including plate preparation, image acquisition and image analysis of zebrafish embryos with approximate timings. (B,C) Brightfield image (B) and fluorescent image (C) of a Tg(itga2b:GFP) zebrafish at 3 dpf acquired from the Hermes showing some of the regions identified by Athena: tail (red), trunk (dark blue), head (white), eye (pink), yolk (yellow), tail fin (light blue) and fluorescent granules (green). (D) Cartoon showing the location of haematopoietic stem and progenitor cells (HSPCs) in the caudal haematopoietic tissue (CHT) in the tail at 3 dpf. (E) Correlation of HSPC counts (between manual and granule) in 93 individual images of Tg(itga2b:GFP) embryos at 3 dpf using the Athena Zebrafish Application, analysed using simple linear regression with r=0.844. Scale bars: 500 µm. n refers to the number of embryos analysed.
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