Single-molecule tracking in living microbial cells  

作　　者：Xiaomin Chen Qianhong Guo Jiexin Guan Lu Zhang Ting Jiang Liping Xie Jun Fan 

机构地区：[1]Institute of Fundamental and Frontier Sciences,University of Electronic Science and Technology of China,Chengdu 611731,China [2]School of Life Sciences and Engineering,Southwest University of Science and Technology,Mianyang 621010,Sichuan,China [3]School of Life Sciences and Technology,University of Electronic Science and Technology of China,Chengdu 611731,China [4]Yangtze Delta Region Institute(Huzhou),University of Electronic Science and Technology of China,Huzhou 313000,Zhejiang,China

出　　处：《Biophysics Reports》2025年第1期1-11,共11页生物物理学报(英文版)

基　　金：supported by the National Key Research and Development Program of China(2023YFC3402400);National Natural Science Foundation of China(32101049,32271271 and 62205048);the Sichuan Science&Technology Program(2023NSFSC0638);NHC Key Laboratory of Nuclear Technology Medical Transformation(Mianyang Central Hospital)(2021HYX007)。

摘　　要：Some microbes are referred to as model organisms because they are easy to study in the laboratory and hold the ability to retain their characteristics during DNA replication,DNA transcription,and other fundamental processes.Studying these microbes in living cells via single-molecule imaging allows us to better understand these processes at highly improved spatiotemporal resolution.Single particle tracking photoactivated localization microscopy(sptPALM)is a robust tool for detecting the positions and motions of individual molecules with tens of nanometers of spatial and millisecond temporal resolution in vivo,providing insights into intricate intracellular environments that traditional ensemble methods cannot.With this approach,the fluorophores are photoactivated stochastically,a series of images are recorded,and the positions of fluorophores are identified in these images,and ultimately the locations are linked together to yield trajectories of individual molecules.Quantitative kinetic and spatial information,such as reaction rates,diffusion coefficients,and localization maps,can be obtained by further analysis.Here,we present a single-molecule tracking protocol that includes sample preparation,data acquisition and brief data processing.This protocol will enable researchers to directly unveil molecular and cellular mechanisms underlying the essential biological processes.

关 键 词：Single-molecule tracking PALM Diffusion coefficients TRAJECTORY Living microbial cells 

分 类 号：Q932[生物学—微生物学]