Super-resolution correlative light-electron microscopy using a click-chemistry approach for studying intracellular trafficking

Teodora Andrian, Thomas Bakkum, Daphne M. van Elsland, Erik Bos, Abraham J. Koster, Lorenzo Albertazzi, Sander I. van Kasteren, Sílvia Pujals

Onderzoeksoutput: Hoofdstuk in Boek/Rapport/CongresprocedureHoofdstukAcademicpeer review

2 Citaten (Scopus)

Samenvatting

Correlative light and electron microscopy (CLEM) entails a group of multimodal imaging techniques that are combined to pinpoint to the location of fluorescently labeled molecules in the context of their ultrastructural cellular environment. Here we describe a detailed workflow for STORM-CLEM, in which STochastic Optical Reconstruction Microscopy (STORM), an optical super-resolution technique, is correlated with transmission electron microscopy (TEM). This protocol has the advantage that both imaging modalities have resolution at the nanoscale, bringing higher synergies on the information obtained. The sample is prepared according to the Tokuyasu method followed by click-chemistry labeling and STORM imaging. Then, after heavy metal staining, electron microscopy imaging is performed followed by correlation of the two images. The case study presented here is on intracellular pathogens, but the protocol is versatile and could potentially be applied to many types of samples.

Originele taal-2Engels
TitelMethods in Cell Biology
RedacteurenThomas Müller-Reichert, Paul Verkade
UitgeverijAcademic Press Inc.
Pagina's303-331
Aantal pagina's29
ISBN van geprinte versie9780128220580
DOI's
StatusGepubliceerd - jan 2021

Publicatie series

NaamMethods in Cell Biology
Volume162
ISSN van geprinte versie0091-679X

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