Danuser Gaudenz, Waterman-Storer Clare M
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Annu Rev Biophys Biomol Struct. 2006;35:361-87. doi: 10.1146/annurev.biophys.35.040405.102114.
Fluorescent speckle microscopy (FSM) is a technology used to analyze the dynamics of macromolecular assemblies in vivo and in vitro. Speckle formation by random association of fluorophores with a macromolecular structure was originally discovered for microtubules. Since then FSM has been expanded to study other cytoskeleton and cytoskeleton-binding proteins. Specialized software has been developed to convert the stochastic speckle image signal into spatiotemporal maps of polymer transport and turnover in living cells. These maps serve as a unique quantitative readout of the dynamic steady state of the cytoskeleton and its responses to molecular and genetic interventions, allowing a systematic study of the mechanisms of cytoskeleton regulation and its effect on cell function. Here, we explain the principles of FSM imaging and signal analysis, outline the biological questions and corresponding methodological advances that have led to the current state of FSM, and give a glimpse of new FSM modalities under development.
荧光斑点显微镜术（FSM）是一种用于分析体内和体外大分子组装体动力学的技术。荧光团与大分子结构随机结合形成斑点最初是在微管中发现的。从那时起，FSM已扩展到研究其他细胞骨架和细胞骨架结合蛋白。已经开发出专门的软件，用于将随机斑点图像信号转换为活细胞中聚合物运输和周转的时空图。这些图是细胞骨架动态稳态及其对分子和基因干预反应的独特定量读数，能够系统地研究细胞骨架调节机制及其对细胞功能的影响。在此，我们解释FSM成像和信号分析的原理，概述导致FSM当前状态的生物学问题和相应的方法学进展，并简要介绍正在开发的新FSM模式。
