Protocol for single-molecule labeling and tracking of bacterial cell division proteins

Di Yan; Jinchan Xue; Jie Xiao; Zhixin Lyu; Xinxing Yang

doi:10.1016/j.xpro.2023.102766

Protocol for single-molecule labeling and tracking of bacterial cell division proteins

STAR Protoc. 2024 Mar 15;5(1):102766. doi: 10.1016/j.xpro.2023.102766. Epub 2023 Dec 12.

Authors

Di Yan¹, Jinchan Xue¹, Jie Xiao², Zhixin Lyu³, Xinxing Yang⁴

Affiliations

¹ School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China.
² Department of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA. Electronic address: [email protected].
³ Department of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA. Electronic address: [email protected].
⁴ School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China. Electronic address: [email protected].

Abstract

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.

Keywords: Microbiology; Microscopy; Single-molecule Assays.

MeSH terms

Bacterial Proteins*
Cell Division
Single Molecule Imaging*

Substances

Bacterial Proteins

Grants and funding

R35 GM136436/GM/NIGMS NIH HHS/United States