Abstract：
Single-molecule tracking (SMT) is a powerful approach to quantify the biophysical parameters of protein dynamics in live cells. Here, we describe a protocol for SMT in live cells of the budding yeast Saccharomyces cerevisiae. We detail how to genetically engineer yeast strains for SMT, how to set up image acquisition parameters, and how different software programs can be used to quantify a variety of biophysical parameters such as diffusion coefficient, residence time, bound fraction, jump angles, and target-search parameters. For complete details on the use and execution of this protocol, please refer to Mehta et al. 1 and Ball et al..2.
摘要：
单分子跟踪（SMT）是量化活细胞中蛋白质动力学的生物物理参数的强大方法。这里,我们描述了在出芽酵母酿酒酵母的活细胞中SMT的方案。我们详细介绍了如何为SMT基因工程酵母菌株，如何设置图像采集参数，以及如何使用不同的软件程序来量化各种生物物理参数，例如扩散系数，停留时间,约束分数，跳跃角度,和目标搜索参数。有关此协议的使用和执行的完整详细信息，请参考Mehta等人。1和Ball等人。.
