Danuser G, Waterman-Storer C M
BioMicrometrics Group, Laboratory for Biomechanics, ETH Zürich, 8952 Schlieren, Switzerland.
J Microsc. 2003 Sep;211(Pt 3):191-207. doi: 10.1046/j.1365-2818.2003.01222.x.
Fluorescent speckle microscopy (FSM) is a technology for analysing the dynamics of macromolecular assemblies. Originally, the effect of random speckle formation was discovered with microtubules. Since then, the method has been expanded to other proteins of the cytoskeleton such as f-actin and microtubule binding proteins. Newly developed, specialized software for analysing speckle movement and photometric fluctuation in the context of polymer transport and turnover has turned FSM into a powerful method for the study of cytoskeletal dynamics in cell migration, division, morphogenesis and neuronal path finding. In all these settings, FSM serves as the quantitative readout to link molecular and genetic interventions to complete maps of the cytoskeleton dynamics and thus can be used for the systematic deciphering of molecular regulation of the cytoskeleton. Fully automated FSM assays can also be applied to live-cell screens for toxins, chemicals, drugs and genes that affect cytoskeletal dynamics. We envision that FSM has the potential to become a core tool in automated, cell-based molecular diagnostics in cases where variations in cytoskeletal dynamics are a sensitive signal for the state of a disease, or the activity of a molecular perturbant. In this paper, we review the origins of FSM, discuss these most recent technical developments and give a glimpse to future directions and potentials of FSM. It is written as a complement to the recent review (Waterman-Storer & Danuser, 2002, Curr. Biol., 12, R633-R640), in which we emphasized the use of FSM in cell biological applications. Here, we focus on the technical aspects of making FSM a quantitative method.
荧光斑点显微镜技术（FSM）是一种用于分析大分子组装体动力学的技术。最初，随机斑点形成的效应是在微管中发现的。从那时起，该方法已扩展到细胞骨架的其他蛋白质，如丝状肌动蛋白和微管结合蛋白。新开发的专门用于分析聚合物运输和周转过程中斑点运动和光度波动的软件，使FSM成为研究细胞迁移、分裂、形态发生和神经元路径寻找中细胞骨架动力学的强大方法。在所有这些情况下，FSM作为定量读数，将分子和基因干预与细胞骨架动力学的完整图谱联系起来，因此可用于系统地解读细胞骨架的分子调控。全自动FSM检测也可应用于针对影响细胞骨架动力学的毒素、化学物质、药物和基因的活细胞筛选。我们设想，在细胞骨架动力学变化是疾病状态或分子扰动剂活性的敏感信号的情况下，FSM有潜力成为基于细胞的自动化分子诊断的核心工具。在本文中，我们回顾了FSM的起源，讨论了这些最新的技术发展，并展望了FSM的未来方向和潜力。本文是对最近一篇综述（Waterman-Storer & Danuser，2002，《当代生物学》，12，R633-R640）的补充，在那篇综述中我们强调了FSM在细胞生物学应用中的使用。在这里，我们专注于使FSM成为定量方法的技术方面。
