Stehbens Samantha J, Wittmann Torsten
Institute of Health Biomedical Innovation (IHBI), Queensland University of Technology Translational Research Institute, Brisbane, Queensland, Australia.
Department of Cell and Tissue Biology, University of California, San Francisco, USA.
Methods Cell Biol. 2014;123:335-46. doi: 10.1016/B978-0-12-420138-5.00018-5.
Recent advances in optical and fluorescent protein technology have rapidly raised expectations in cell biology, allowing quantitative insights into dynamic intracellular processes like never before. However, quantitative live-cell imaging comes with many challenges including how best to translate dynamic microscopy data into numerical outputs that can be used to make meaningful comparisons rather than relying on representative data sets. Here, we use analysis of focal adhesion turnover dynamics as a straightforward specific example on how to image, measure, and analyze intracellular protein dynamics, but we believe this outlines a thought process and can provide guidance on how to understand dynamic microcopy data of other intracellular structures.
光学和荧光蛋白技术的最新进展迅速提升了细胞生物学领域的期望,使人们能够以前所未有的方式对细胞内动态过程进行定量洞察。然而,定量活细胞成像面临诸多挑战,包括如何将动态显微镜数据转化为可用于进行有意义比较的数值输出,而不是依赖代表性数据集。在此,我们以粘着斑周转动力学分析为例,直观地说明如何对细胞内蛋白质动力学进行成像、测量和分析,但我们认为这勾勒出了一种思维过程,并可为理解其他细胞内结构的动态显微镜数据提供指导。