这里,我们提出了一个标记和跟踪单个分子的协议,特别是活细菌细胞中的细胞分裂蛋白。协议包括应变构造,单分子成像,轨迹分割,和运动属性分析。该方案能够识别与不同细胞分裂蛋白相关的独特运动状态。随后对这些蛋白质的动态行为的评估提供了对它们在细胞分裂过程中在隔膜处的活性和相互作用的见解。有关此协议的使用和执行的完整详细信息,请参考杨等人。(2021),1Lyu等人。(2022),2和Mahone等人。(2024).3.
Here, we present a protocol for labeling and tracking individual molecules, particularly cell division proteins in live bacterial cells. The protocol encompasses strain construction, single-molecule imaging, trajectory segmentation, and motion property analysis. The protocol enables the identification of distinctive motion states associated with different cell division proteins. Subsequent assessments of the dynamic behaviors of these proteins provide insights into their activities and interactions at the septum during cell division. For complete details on the use and execution of this protocol, please refer to Yang et al. (2021),1 Lyu et al. (2022),2 and Mahone et al. (2024).3.