1. Ultrastructural Imaging of Salmonella -Host Interactions Using Super-resolution Correlative Light-Electron Microscopy of Bioorthogonal Pathogens
- Author
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Lorenzo Albertazzi, Thomas Bakkum, Nikolaos Oikonomeas-Koppasis, Daphne M. van Elsland, Abraham J. Koster, Ilana Berlin, Jacques Neefjes, Silvia Pujals, Erik Bos, Annemarie H. Meijer, Sander I. van Kasteren, and Molecular Biosensing for Med. Diagnostics
- Subjects
0301 basic medicine ,Context (language use) ,010402 general chemistry ,01 natural sciences ,Biochemistry ,law.invention ,03 medical and health sciences ,law ,Microscopy ,Fluorescence microscope ,host-pathogen interactions ,Molecular Biology ,Full Paper ,electron microscopy ,Chemistry ,Intracellular parasite ,Organic Chemistry ,Resolution (electron density) ,Full Papers ,bioorthogonal chemistry ,infection ,0104 chemical sciences ,030104 developmental biology ,Biophysics ,Ultrastructure ,Molecular Medicine ,fluorescence ,Electron microscope ,Bioorthogonal chemistry ,host–pathogen interactions - Abstract
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.
- Published
- 2018
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