Mito Mari, Kawaguchi Tetsuya, Hirose Tetsuro, Nakagawa Shinichi
RNA Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako 351-0198, Japan.
Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
Methods. 2016 Apr 1;98:158-165. doi: 10.1016/j.ymeth.2015.11.007. Epub 2015 Nov 10.
A number of non-membranous cellular bodies have been identified in higher eukaryotes, and these bodies contain a specific set of proteins and RNAs that are used to fulfill their functions. The size of these RNA-containing cellular bodies is usually on a submicron scale, making it difficult to observe fine structures using optical microscopy due to the diffraction limitation of visible light. Recently, microscope companies have released super-resolution microscopes that were developed using different principles, enabling the observation of sub-micron structures not resolvable in conventional fluorescent microscopy. Here, we describe multi-color fluorescent in situ hybridization techniques optimized for the simultaneous detection of RNA and proteins using super-resolution microscopy, namely structured illumination microscopy (SIM).
在高等真核生物中已鉴定出许多非膜性细胞体，这些细胞体包含一组特定的蛋白质和RNA，用于履行其功能。这些含RNA的细胞体大小通常在亚微米尺度，由于可见光的衍射限制，使用光学显微镜难以观察到精细结构。最近，显微镜公司推出了基于不同原理开发的超分辨率显微镜，能够观察到传统荧光显微镜无法分辨的亚微米结构。在此，我们描述了使用超分辨率显微镜（即结构照明显微镜，SIM）同时检测RNA和蛋白质而优化的多色荧光原位杂交技术。
