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Ultrastructural Imaging of Salmonella –Host Interactions Using Super‐resolution Correlative Light‐Electron Microscopy of Bioorthogonal Pathogens
Author(s) -
van Elsland Daphne M.,
Pujals Sílvia,
Bakkum Thomas,
Bos Erik,
OikonomeasKoppasis Nikolaos,
Berlin Ilana,
Neefjes Jacques,
Meijer Annemarie H.,
Koster Abraham J.,
Albertazzi Lorenzo,
van Kasteren Sander I.
Publication year - 2018
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800230
Subject(s) - bioorthogonal chemistry , context (archaeology) , microscopy , ultrastructure , intracellular parasite , biology , intracellular , electron microscope , salmonella , pathogen , fluorescence microscope , correlative , microbiology and biotechnology , biophysics , chemistry , bacteria , fluorescence , optics , physics , anatomy , genetics , click chemistry , paleontology , linguistics , philosophy , combinatorial chemistry
The imaging of intracellular pathogens inside host cells is complicated by the low resolution and sensitivity of fluorescence microscopy and by the lack of ultrastructural information to visualize the pathogens. Herein, we present a new method to visualize these pathogens during infection that circumvents these problems: by using a metabolic hijacking approach to bioorthogonally label the intracellular pathogen Salmonella Typhimurium and by using these bioorthogonal groups to introduce fluorophores compatible with stochastic optical reconstruction microscopy (STORM) and placing this in a correlative light electron microscopy (CLEM) workflow, the pathogen can be imaged within its host cell context Typhimurium with a resolution of 20 nm. This STORM‐CLEM approach thus presents a new approach to understand these pathogens during infection.