A versatile, automated and high-throughput drug screening platform for zebrafish embryos.
A versatile, automated and high-throughput drug screening platform for zebrafish embryos.
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DOI：
10.1242/bio.058513
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发表时间：
2021-09-15
期刊：
影响因子：
2.4
通讯作者：
Payne E
中科院分区：
文献类型：
--
作者：
Lubin A;Otterstrom J;Hoade Y;Bjedov I;Stead E;Whelan M;Gestri G;Paran Y;Payne E
斑马鱼为活体动物的药物筛选提供了一个独特的机会，快速发育的透明胚胎允许相对高通量的显微镜筛选。然而，快速、灵活的成像和分析平台的有限可用性限制了斑马鱼在药物筛选中的使用。我们开发了一种易于使用、可定制的自动化筛选程序，适用于高通量基于表型的活斑马鱼筛选。我们利用WiScan®爱马仕高内容成像系统快速获取胚胎的明场和荧光图像，并利用WiSoft® Athena Zebrafish应用程序进行分析，该应用程序利用人工智能驱动的算法自动检测明场图像中的鱼类，识别解剖结构，将动物划分为区域并专门选择所需的侧向鱼类。我们最初的验证结合了荧光图像的结构分析，以计数胚胎尾部的GFP标记的造血干细胞和祖细胞，这与手动计数相关。我们进一步验证了该系统，以评估基因突变和X射线照射的影响，在高含量使用广泛的测定。此外，我们使用双荧光团以高通量对多种细胞类型进行了同时分析。总之，我们展示了一个广泛适用且可快速定制的平台，用于斑马鱼的高内容筛选。 这篇文章有一个相关的第一人称采访的第一作者。总结：我们开发并验证了一个快速，灵活和可快速定制的平台，自动化图像采集和定量分析，用于活斑马鱼胚胎中的高含量药物筛选。
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 . Summary: We developed and validated a fast, flexible and rapidly customisable platform, automating both image acquisition and quantitative analysis, for high-content drug screening in live zebrafish embryos.
DOI：
10.1016/j.ddmod.2012.02.001
发表时间：
2013
期刊：
Drug discovery today. Disease models
影响因子：
--
作者：
Esterberg, Robert;Coffin, Allison B;Ou, Henry;Simon, Julian A;Raible, David W;Rubel, Edwin W
通讯作者：
Rubel, Edwin W