Mennella Vito, Hanna Rachel, Kim Moshe
Department of Biochemistry, University of Toronto, Toronto, ON, Canada; Cell Biology Program, The Hospital for Sick Children, Toronto, ON, Canada; Peter Gilgan Centre for Research and Learning, Toronto, ON, Canada.
Department of Biochemistry, University of Toronto, Toronto, ON, Canada; Cell Biology Program, The Hospital for Sick Children, Toronto, ON, Canada.
Methods Cell Biol. 2015;129:129-152. doi: 10.1016/bs.mcb.2015.03.009. Epub 2015 May 27.
In this chapter, we describe the current methods of examining the structure of centrosomes by fluorescence subdiffraction microscopy. By using recently developed microscopy techniques, centrosomal proteins can now be examined in cells with a resolution of only a few nanometers, a level of molecular detail beyond the reach of traditional cell biology methods as confocal and widefield microscopy. We emphasize imaging by three-dimensional structured illumination microscopy, stochastic optical reconstruction microscopy, and quantitative approaches to image data analysis.
在本章中，我们描述了通过荧光亚衍射显微镜检查中心体结构的当前方法。利用最近开发的显微镜技术，现在可以在细胞中以仅几纳米的分辨率检查中心体蛋白，这是传统细胞生物学方法（如共聚焦显微镜和宽场显微镜）无法企及的分子细节水平。我们重点介绍三维结构照明显微镜、随机光学重建显微镜成像以及图像数据分析的定量方法。
