Ellett Felix, Irimia Daniel
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital-Harvard Medical School-Shriners Burns Hospital;
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital-Harvard Medical School-Shriners Burns Hospital.
J Vis Exp. 2017 Dec 8(130):56498. doi: 10.3791/56498.
Zebrafish have emerged as a powerful model of various human diseases and a useful tool for an increasing range of experimental studies, spanning fundamental developmental biology through to large-scale genetic and chemical screens. However, many experiments, especially those related to infection and xenograft models, rely on microinjection and imaging of embryos and larvae, which are laborious techniques that require skill and expertise. To improve the precision and throughput of current microinjection techniques, we developed a series of microstructured devices to orient and stabilize zebrafish embryos at 2 days post fertilization (dpf) in ventral, dorsal, or lateral orientation prior to the procedure. To aid in the imaging of embryos, we also designed a simple device with channels that orient 4 zebrafish laterally in parallel against a glass cover slip. Together, the tools that we present here demonstrate the effectiveness of photolithographic approaches to generate useful devices for the optimization of zebrafish techniques.
斑马鱼已成为多种人类疾病的强大模型，也是越来越多实验研究的有用工具，涵盖从基础发育生物学到大规模遗传和化学筛选等广泛领域。然而，许多实验，尤其是那些与感染和异种移植模型相关的实验，依赖于胚胎和幼体的显微注射和成像，这些都是费力的技术，需要技巧和专业知识。为了提高当前显微注射技术的精度和通量，我们开发了一系列微结构装置，用于在受精后2天（dpf）将斑马鱼胚胎以腹侧、背侧或侧位定向并稳定，以便在操作前进行处理。为了辅助胚胎成像，我们还设计了一种简单的装置，其通道可将4条斑马鱼横向平行地靠在玻璃盖玻片上。总之，我们在此展示的工具证明了光刻方法在生成用于优化斑马鱼技术的有用装置方面的有效性。
