Yu Tianyu, Jiang Yue, Lin Sijie
College of Environmental Science and Engineering, State Key Laboratory of Pollution Control and Resource Reuse, Tongji University; UNEP-Tongji Institute of Environment for Sustainable Development, Tongji University.
College of Environmental Science and Engineering, State Key Laboratory of Pollution Control and Resource Reuse, Tongji University.
J Vis Exp. 2018 Jun 1(136):57892. doi: 10.3791/57892.
The zebrafish is a globally recognized fresh water organism frequently used in developmental biology, environmental toxicology, and human disease related research fields. Thanks to its unique features, including large fecundity, embryo translucency, rapid and simultaneous development, etc., zebrafish embryos are often used for large scale toxicity assessment of chemicals and drug/compound screening. A typical screening procedure involves adult zebrafish spawning, embryos selection, and arraying the embryos into multi-well plates. From there, embryos are subjected to exposure and the toxicity of chemical, or the effectiveness of the drugs/compounds can be evaluated relatively quickly based on phenotypic observations. Among these processes, embryos arraying is one of the most time-consuming and labor-intensive steps that limits the throughput level. In this protocol, we present an innovative approach that makes use of a 3D-printed arraying template coupled with vacuum manipulation to speed up this laborious step. The protocol herein describes the overall design of the arraying template, a detailed experimental setup and step-by-step procedure, followed by representative results. When implemented, this approach should prove beneficial in a variety of research applications using zebrafish embryos as testing subjects.
斑马鱼是一种全球公认的淡水生物，常用于发育生物学、环境毒理学以及与人类疾病相关的研究领域。由于其具有独特的特征，包括繁殖力强、胚胎半透明、发育迅速且同步等，斑马鱼胚胎常被用于化学品的大规模毒性评估以及药物/化合物筛选。典型的筛选程序包括成年斑马鱼产卵、胚胎选择以及将胚胎排列到多孔板中。从那时起，胚胎接受暴露处理，基于表型观察可以相对快速地评估化学品的毒性或药物/化合物的有效性。在这些过程中，胚胎排列是最耗时且劳动强度最大的步骤之一，限制了通量水平。在本方案中，我们提出了一种创新方法，利用3D打印的排列模板结合真空操作来加速这一繁琐步骤。本文的方案描述了排列模板的总体设计、详细的实验设置和逐步操作步骤，随后展示了代表性结果。当实施时，这种方法在以斑马鱼胚胎作为测试对象的各种研究应用中应会被证明是有益的。
