Department of Molecular Cell Biology, Institute of Biology, Leiden University, The Netherlands.
Methods. 2013 Aug 15;62(3):246-54. doi: 10.1016/j.ymeth.2013.06.002. Epub 2013 Jun 11.
The increasing use of zebrafish larvae for biomedical research applications is resulting in versatile models for a variety of human diseases. These models exploit the optical transparency of zebrafish larvae and the availability of a large genetic tool box. Here we present detailed protocols for the robotic injection of zebrafish embryos at very high accuracy with a speed of up to 2000 embryos per hour. These protocols are benchmarked for several applications: (1) the injection of DNA for obtaining transgenic animals, (2) the injection of antisense morpholinos that can be used for gene knock-down, (3) the injection of microbes for studying infectious disease, and (4) the injection of human cancer cells as a model for tumor progression. We show examples of how the injected embryos can be screened at high-throughput level using fluorescence analysis. Our methods open up new avenues for the use of zebrafish larvae for large compound screens in the search for new medicines.
越来越多的研究人员将斑马鱼幼鱼应用于生物医学研究，这为各种人类疾病提供了多样化的模型。这些模型利用了斑马鱼幼鱼的光学透明性和大量的遗传工具包。在这里，我们提供了详细的协议，用于以高达 2000 个胚胎/小时的速度进行高精度的斑马鱼胚胎机器人注射。这些方案在以下几个方面进行了基准测试：（1）注射 DNA 以获得转基因动物，（2）注射反义 morpholino 以进行基因敲低，（3）注射微生物以研究传染病，以及（4）注射人类癌细胞作为肿瘤进展的模型。我们展示了如何使用荧光分析在高通量水平上对注射后的胚胎进行筛选的示例。我们的方法为使用斑马鱼幼鱼进行大规模化合物筛选以寻找新药开辟了新途径。
