Reck-Peterson Samara L, Derr Nathan D, Stuurman Nico
Cold Spring Harb Protoc. 2010 Mar;2010(3):pdb.prot5399. doi: 10.1101/pdb.prot5399.
Total internal reflection fluorescence microscopy (TIRFM) allows fluorescent molecules on or near the plasma membrane to be visualized with a very high signal-to-noise ratio. This strategy has been very successful for imaging molecular machines as they move and do work. We provide here a general protocol for imaging single molecular motors as they move along microtubule tracks. Our protocol is designed for the study of cytoplasmic dynein purified from Saccharomyces cerevisiae, but it represents a general framework for any in vitro single-molecule assay.
全内反射荧光显微镜（TIRFM）能够以非常高的信噪比观察质膜上或其附近的荧光分子。这种策略在对分子机器的运动及工作过程进行成像方面非常成功。我们在此提供一种通用方案，用于对沿着微管轨道移动的单个分子马达进行成像。我们的方案是为研究从酿酒酵母中纯化的细胞质动力蛋白而设计的，但它代表了任何体外单分子检测的通用框架。
