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A simple procedure to analyze positions of interest in infectious cell cultures by correlative light and electron microscopy.

Authors :
Madela K
Banhart S
Zimmermann A
Piesker J
Bannert N
Laue M
Source :
Methods in cell biology [Methods Cell Biol] 2014; Vol. 124, pp. 93-110.
Publication Year :
2014

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.<br /> (© 2014 Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
0091-679X
Volume :
124
Database :
MEDLINE
Journal :
Methods in cell biology
Publication Type :
Academic Journal
Accession number :
25287838
Full Text :
https://doi.org/10.1016/B978-0-12-801075-4.00005-7