Metabolic biorthogonal labeling and dSTORM imaging of peptidoglycan synthesis in Streptococcus pneumoniae

Jennyfer Trouve; Oleksandr Glushonkov; Cecile Morlot

doi:10.1016/j.xpro.2021.101006

Metabolic biorthogonal labeling and dSTORM imaging of peptidoglycan synthesis in Streptococcus pneumoniae

STAR Protoc. 2021 Dec 13;2(4):101006. doi: 10.1016/j.xpro.2021.101006. eCollection 2021 Dec 17.

Authors

Jennyfer Trouve¹, Oleksandr Glushonkov¹, Cecile Morlot¹

Affiliation

¹ Univ. Grenoble Alpes, CNRS, CEA, IBS, 38000 Grenoble, France.

Abstract

Fluorescence microscopy is a method of choice for studying peptidoglycan assembly, but it presents two major challenges: the peptidoglycan must be labeled with a probe that will not perturb the physiological process, and the spatial resolution must reach the nanometer scale to reveal fine details of the synthesis process. This protocol meets both challenges by combining biorthogonal metabolic labeling of peptidoglycan in Streptococcus pneumoniae with super-resolution fluorescence microscopy (dSTORM), also providing cues to adapt it to other bacteria. For complete details on the use and execution of this protocol, please refer to Trouve et al. (2021).

Keywords: Cell Biology; Chemistry; Microbiology; Microscopy; Molecular/Chemical Probes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Female
Male
Mice
Mice, Inbred C57BL
Microscopy, Fluorescence / methods*
Molecular Imaging / methods*
Peptidoglycan* / analysis
Peptidoglycan* / chemistry
Peptidoglycan* / metabolism
Streptococcus pneumoniae* / chemistry
Streptococcus pneumoniae* / metabolism

Substances

Peptidoglycan