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Ultrastructural Imaging of Salmonella–Host Interactions Using Super‐resolution Correlative Light‐Electron Microscopy of Bioorthogonal Pathogens |
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| Authors: | Dr. Daphne M. van Elsland Dr. Sílvia Pujals Thomas Bakkum Dr. Erik Bos Nikolaos Oikonomeas‐Koppasis Dr. Ilana Berlin Dr. Jacques Neefjes Dr. Annemarie H. Meijer Dr. Abraham J. Koster Dr. Lorenzo Albertazzi Dr. Sander I. van Kasteren |
| Affiliation: | 1. Leiden Institute of Chemistry and, The Institute for Chemical Immunology, Leiden University, Leiden, The Netherlands;2. Department of Cell and Chemical Biology, Institute for Chemical Immunology, Leiden University Medical Center LUMC, Leiden, The Netherlands;3. Department of Nanoscopy for Nanomedicine, Institute of Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, Barcelona, Spain;4. Department of Electron Microscopy, Leiden University Medical Center LUMC, Leiden, The Netherlands;5. Institute of Biology Leiden, Leiden University, Leiden, The Netherlands;6. Department of Biomedical Engineering and Institute of Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands |
| 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. |
| Keywords: | bioorthogonal chemistry electron microscopy fluorescence host– pathogen interactions infection |