Correlative light and electron microscopy is an imaging technique that enables identification and targeting of fluorescently tagged structures with subsequent imaging at near-to-nanometer resolution. We established a novel correlative cryo-fluorescence microscopy and cryo-scanning electron microscopy workflow, which enables imaging of the studied object of interest very close to its natural state, devoid of artifacts caused for instance by slow chemical fixation. This system was tested by investigating the interaction of the zoonotic bacterium Borrelia burgdorferi with two mammalian cell lines of neural origin in order to broaden our knowledge about the cell-association mechanisms that precedes the entry of the bacteria into the cell. This method appears to be an unprecedentedly fast (<3 hours), straightforward, and reliable solution to study the finer details of pathogen-host cell interactions and provides important insights into the complex and dynamic relationship between a pathogen and a host.

Department of Virology Veterinary Research Institute Brno CZ 62100 Czech Republic

Faculty of Science Charles University Prague Viničná 1594 7 Praha CZ 12800 Czech Republic

Faculty of Science University of South Bohemia in České Budějovice Branišovská 1760 České Budějovice CZ 37005 Czech Republic

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Branišovská 31 České Budějovice CZ 37005 Czech Republic

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Pathogenicity and virulence of Borrelia burgdorferi

Cathodoluminescence imaging of cellular structures labeled with luminescent iridium or rhenium complexes at cryogenic temperatures

Novel targets and strategies to combat borreliosis

Pleomorphism and Viability of the Lyme Disease Pathogen Borrelia burgdorferi Exposed to Physiological Stress Conditions: A Correlative Cryo-Fluorescence and Cryo-Scanning Electron Microscopy Study