Jiang Yuxiang, Huang Shanjin
Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing, China.
Institute of Botany, Chinese Academy of Sciences, Beijing, China.
Bio Protoc. 2017 Mar 5;7(5):e2146. doi: 10.21769/BioProtoc.2146.
Total internal reflection fluorescence (TIRF) microscopy is a powerful tool for visualizing the dynamics of actin filaments at single-filament resolution . Thanks to the development of various fluorescent probes, we can easily monitor all kinds of events associated with actin dynamics, including nucleation, elongation, bundling, fragmentation and monomer dissociation. Here we present a detailed protocol regarding the visualization and quantification of actin nucleation and filament elongation events by TIRF microscopy , which is based on the methods previously reported ( Liu , 2015 ; Yang , 2011 ).
全内反射荧光（TIRF）显微镜是一种强大的工具，可在单丝分辨率下可视化肌动蛋白丝的动力学。由于各种荧光探针的发展，我们可以轻松监测与肌动蛋白动力学相关的各种事件，包括成核、伸长、成束、断裂和单体解离。在这里，我们基于先前报道的方法（Liu，2015；Yang，2011），给出了一份关于通过TIRF显微镜可视化和定量肌动蛋白成核和丝伸长事件的详细方案。
