Universiteit Leiden

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Dissertation

Bioorthogonal Labeling Tools to Study Pathogenic Intracellular Bacteria

In this thesis, bioorthogonal chemistry is combined with correlative light-electron microscopy to selectively label and study pathogenic intracellular bacteria within the host immune cell.

Author
Bakkum, T.
Date
17 November 2021
Links
Thesis in Leiden Repository

In this thesis, bioorthogonal chemistry is combined with correlative light-electron microscopy to selectively label and study pathogenic intracellular bacteria within the host immune cell. This technique combines the ultrastructural information of transmission electron microscopy with the functional information of fluorescence light microscopy in order to investigate the host-pathogen interactions that contribute to the diseases caused by pathogenic intracellular bacteria such as Salmonella Typhimurium and Mycobacterium tuberculosis. The technique is further expanded with super-resolution microscopy by combining stochastic optical reconstruction microscopy with transmission electron microscopy. Additionally, the bioorthogonal labeling method for the study of intracellular bacteria is validated through a bead-based stability assay, demonstrating the compatibility of alkyne and azide groups to label bacterial proteins within the degradative lysosomal environment. The technique developed in this thesis may contribute to a better understanding of the mechanisms behind bacterial diseases, as well as the development of novel antibiotics and other therapies to fight these important infectious diseases.

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