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

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Title of host publicationCorrelative Light and Electron Microscopy IV
EditorsThomas Müller-Reichert, Paul Verkade
PublisherElsevier
Chapter13
Pages303-331
Number of pages29
ISBN (Print)9780128220580
DOIs
Publication statusPublished - Jan 2021

Publication series

NameMethods in Cell Biology
Volume162
ISSN (Print)0091-679X

Funding

This work was financially supported by the Ministerio de Economía y Competitividad/Agencia Estatal de Investigación (AEI) (NANOVAX: EURONANOMED/Acciones de Programación Conjunta Internacional PCIN-2016-025), Spanish Ministry of Science and Innovation (PID2019-109450RB-I00/AEI/ 10.13039/501100011033 ), European Research Council/Horizon 2020 (ERC-StG-757,397), “la Caixa” Foundation (ID 100010434), and by the Generalitat de Catalunya (2017 SGR 01536) to S.P. and L.A, and NWO Echo (711.015.008) to T.B. and S.I.K. T.A. was supported by a Marie Skłodowska-Curie Horizon 2020 (713673), “la Caixa” Foundation (LCF/BQ/DI18/11660039) fellowship. S.P. is grateful to Prof. J. E. Heuser for his patience and guidance in introducing her to the electron microscopy world.

FundersFunder number
European Union's Horizon 2020 - Research and Innovation Framework ProgrammeERC-StG-757,397
Marie Skłodowska‐CurieLCF/BQ/DI18/11660039
“la Caixa” Foundation100010434
European Union's Horizon 2020 - Research and Innovation Framework Programme713673
Generalitat de Catalunya2017 SGR 01536
Nederlandse Organisatie voor Wetenschappelijk Onderzoek711.015.008
Ministerio de Ciencia e InnovaciónPID2019-109450RB-I00/AEI/ 10.13039/501100011033

    Keywords

    • Click-chemistry
    • Correlative light and electron microscopy
    • Single molecule localization microscopy
    • STochastic Optical Reconstruction Microscopy (STORM)
    • Super-resolution microscopy
    • Tokuyasu cryo-sectioning
    • Transmission electron microscopy

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