Toya Mika, Iida Yumi, Sugimoto Asako
Laboratory for Developmental Genomics, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Methods Cell Biol. 2010;97:359-72. doi: 10.1016/S0091-679X(10)97019-2.
Development of the nematode Caenorhabditis elegans is highly reproducible, and the cell division patterns are virtually invariant. Transparency of the eggshell and cells enables the observation of intracellular events with a high temporal and spatial resolution. These unique features, along with the sophisticated genetic techniques, make this organism one of the most attractive model systems for dissecting regulatory mechanisms of dynamic cellular behaviors, such as mitosis, at an organismal level. In this chapter, we describe immunofluorescence and live imaging methods for analyzing mitotic spindle regulation. In particular, we present the use of double- or triple-labeled fluorescent strains for high-resolution two-dimensional and three-dimensional live imaging to analyze dynamic behaviors of mitotic spindles.
线虫秀丽隐杆线虫的发育具有高度的可重复性，其细胞分裂模式几乎是不变的。卵壳和细胞的透明性使得能够以高时空分辨率观察细胞内事件。这些独特的特征，连同复杂的遗传技术，使这种生物体成为在生物体水平上剖析动态细胞行为（如有丝分裂）调控机制最具吸引力的模型系统之一。在本章中，我们描述了用于分析有丝分裂纺锤体调控的免疫荧光和实时成像方法。特别是，我们介绍了使用双标记或三标记荧光菌株进行高分辨率二维和三维实时成像，以分析有丝分裂纺锤体的动态行为。
