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首页> 美国卫生研究院文献>PLoS Clinical Trials >Miniaturized Embryo Array for Automated Trapping, Immobilization and Microperfusion of Zebrafish Embryos
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Miniaturized Embryo Array for Automated Trapping, Immobilization and Microperfusion of Zebrafish Embryos

机译：小型化的胚胎阵列，用于斑马鱼胚胎的自动诱捕，固定和微灌注

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Zebrafish (Danio rerio) has recently emerged as a powerful experimental model in drug discovery and environmental toxicology. Drug discovery screens performed on zebrafish embryos mirror with a high level of accuracy the tests usually performed on mammalian animal models, and fish embryo toxicity assay (FET) is one of the most promising alternative approaches to acute ecotoxicity testing with adult fish. Notwithstanding this, automated in-situ analysis of zebrafish embryos is still deeply in its infancy. This is mostly due to the inherent limitations of conventional techniques and the fact that metazoan organisms are not easily susceptible to laboratory automation. In this work, we describe the development of an innovative miniaturized chip-based device for the in-situ analysis of zebrafish embryos. We present evidence that automatic, hydrodynamic positioning, trapping and long-term immobilization of single embryos inside the microfluidic chips can be combined with time-lapse imaging to provide real-time developmental analysis. Our platform, fabricated using biocompatible polymer molding technology, enables rapid trapping of embryos in low shear stress zones, uniform drug microperfusion and high-resolution imaging without the need of manual embryo handling at various developmental stages. The device provides a highly controllable fluidic microenvironment and post-analysis eleuthero-embryo stage recovery. Throughout the incubation, the position of individual embryos is registered. Importantly, we also for first time show that microfluidic embryo array technology can be effectively used for the analysis of anti-angiogenic compounds using transgenic zebrafish line (fli1a:EGFP). The work provides a new rationale for rapid and automated manipulation and analysis of developing zebrafish embryos at a large scale.
机译：斑马鱼（Danio rerio）最近在药物发现和环境毒理学中成为强大的实验模型。在斑马鱼胚胎镜上进行的药物发现筛选具有很高的准确性，通常可以在哺乳动物动物模型上进行测试，而鱼胚胎毒性试验（FET）是成年鱼急性生态毒性测试中最有希望的替代方法之一。尽管如此，斑马鱼胚胎的自动原位分析仍处于起步阶段。这主要是由于常规技术的固有局限性以及后生动物不容易受到实验室自动化的影响。在这项工作中，我们描述了一种新颖的基于芯片的微型化设备的开发，该设备用于斑马鱼胚胎的原位分析。我们目前的证据表明，微流体芯片内部单个胚胎的自动，流体动力学定位，诱集和长期固定可以与延时成像相结合，以提供实时的发育分析。我们的平台是使用生物相容性聚合物成型技术制造的，能够在低剪切应力区域中快速捕获胚胎，实现均匀的药物微灌流和高分辨率成像，而无需在各个发育阶段进行手动胚胎处理。该设备提供了高度可控的流体微环境和分析后的胚胎发育阶段恢复。在整个孵育过程中，将记录单个胚胎的位置。重要的是，我们也首次证明微流体胚胎阵列技术可以有效地用于使用转基因斑马鱼品系（fli1a：EGFP）分析抗血管生成化合物。这项工作为大规模开发斑马鱼胚胎的快速，自动操作和分析提供了新的原理。

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