Correlative cryo-fluorescence and cryo-scanning electron microscopy as a straightforward tool to study host-pathogen interactions

0453307 - BC 2016 RIV GB eng J - Journal Article
Strnad, Martin - Elsterová, Jana - Schrenková, Jana - Vancová, Marie - Rego, Ryan O. M. - Grubhoffer, Libor - Nebesářová, Jana
Correlative cryo-fluorescence and cryo-scanning electron microscopy as a straightforward tool to study host-pathogen interactions.
Scientific Reports. Roč. 5, DEC 10 2015 (2015), s. 18029. ISSN 2045-2322. E-ISSN 2045-2322
R&D Projects: GA TA ČR(CZ) TE01020118; GA ČR GAP502/11/2116
EU Projects: European Commission(XE) 278976; European Commission(XE) 602272
Institutional support: RVO:60077344
Keywords : Borrelia burgdorferi * cryoelectron tomography * lyme disease
Subject RIV: EE - Microbiology, Virology
Impact factor: 5.228, year: 2015

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.
Permanent Link: http://hdl.handle.net/11104/0254166