title: A simple procedure to analyze positions of interest in infectious cell cultures by correlative light and electron microscopy.
abstract: Plastic cell culture dishes that contain a thin bottom of highest optical quality including an imprinted finder grid (μ-Dish Grid-500) are optimally suited for routine correlative light and electron microscopy using chemical fixation. Such dishes allow high-resolution fluorescence and bright-field imaging using fixed and living cells and are compatible with standard protocols for scanning and transmission electron microscopy. Ease of use during cell culture and imaging, as well as a tight cover render the dishes particularly suitable for working with infectious organisms up to the highest biosafety level. Detailed protocols are provided and demonstrated by showing two examples: monitoring the production of virus-like particles of the Human Endogenous Retrovirus HERV-K(HML-2) by HeLa cells and investigation of Rab11-positive membrane-compartments of HeLa cells after infection with Chlamydia trachomatis.
authors.affiliation: Advanced Light and Electron Microscopy (ZBS 4), Robert Koch Institute, Berlin, Germany.
authors.forename: Kazimierz
authors.initials: K
authors.lastname: Madela
authors.affiliation: Junior Research Group Sexually Transmitted Bacterial Pathogens (NG 5), Robert Koch Institute, Berlin, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Banhart
authors.affiliation: Centre for HIV and other Retroviruses (FG 18), Robert Koch Institute, Berlin, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Zimmermann
authors.affiliation: Advanced Light and Electron Microscopy (ZBS 4), Robert Koch Institute, Berlin, Germany.
authors.forename: Janett
authors.initials: J
authors.lastname: Piesker
authors.affiliation: Centre for HIV and other Retroviruses (FG 18), Robert Koch Institute, Berlin, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Bannert
authors.affiliation: Advanced Light and Electron Microscopy (ZBS 4), Robert Koch Institute, Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Laue
mesh_terms: D002692:Chlamydia trachomatis / Q000502:physiology; D022162:Cytoplasmic Vesicles / Q000378:metabolism / Q000648:ultrastructure; D020077:Endogenous Retroviruses / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D008855:Microscopy, Electron, Scanning / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D008867:Microtomy; D059010:Single-Cell Analysis / Q000379:methods*; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
chemical_list: D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins
keywords: CLEM; Cell culture; Chlamydia; Fluorescence microscopy; Infection; Live-cell imaging; Retrovirus; Scanning electron microscopy; Thin section electron microscopy; μ-Dish