A versatile, automated and high-throughput drug screening platform for zebrafish embryos
Alexandra Lubin, Jason Otterstrom, Yvette Hoade, Ivana Bjedov, Eleanor Stead, Matthew Whelan, Gaia Gestri, Yael Paran, Elspeth Payne, Alexandra Lubin, Jason Otterstrom, Yvette Hoade, Ivana Bjedov, Eleanor Stead, Matthew Whelan, Gaia Gestri, Yael Paran, Elspeth Payne
Abstract
Zebrafish provide a unique opportunity for drug screening in living animals, with the fast-developing, transparent embryos allowing for relatively high-throughput, microscopy-based screens. However, the limited availability of rapid, flexible imaging and analysis platforms has limited the use of zebrafish in drug screens. We have developed an easy-to-use, customisable automated screening procedure suitable for high-throughput phenotype-based screens of live zebrafish. We utilised the WiScan® Hermes High Content Imaging System to rapidly acquire brightfield and fluorescent images of embryos, and the WiSoft® Athena Zebrafish Application for analysis, which harnesses an Artificial Intelligence-driven algorithm to automatically detect fish in brightfield images, identify anatomical structures, partition the animal into regions and exclusively select the desired side-oriented fish. Our initial validation combined structural analysis with fluorescence images to enumerate GFP-tagged haematopoietic stem and progenitor cells in the tails of embryos, which correlated with manual counts. We further validated this system to assess the effects of genetic mutations and X-ray irradiation in high content using a wide range of assays. Further, we performed simultaneous analysis of multiple cell types using dual fluorophores in high throughput. In summary, we demonstrate a broadly applicable and rapidly customisable platform for high-content screening in zebrafish. This article has an associated First Person interview with the first author of the paper.
Keywords: Drug screening; High-throughput; Zebrafish.
Conflict of interest statement
Competing interests J.O. and Y.P. are employed by IDEA Bio-Medical, Rehovot, Israel, which manufactures and sells the WiScan® Hermes High Content Imaging System and WiSoft® Athena software Zebrafish Application used throughout this study. They have provided technical support throughout development of this screening platform and have contributed to the preparation of the manuscript. M.W. is employed by University College London with a fellowship partially funded by IDEA Bio-Medical. The remaining authors have no competing interests.
© 2021. Published by The Company of Biologists Ltd.
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References
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