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

doi:10.1016/bs.mcb.2020.09.001

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

Methods Cell Biol. 2021:162:303-331. doi: 10.1016/bs.mcb.2020.09.001. Epub 2020 Oct 16.

Authors

Teodora Andrian¹, Thomas Bakkum², Daphne M van Elsland³, Erik Bos⁴, Abraham J Koster⁴, Lorenzo Albertazzi⁵, Sander I van Kasteren⁶, Sílvia Pujals⁷

Affiliations

¹ Institute of Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, Barcelona, Spain.
² Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Leiden, The Netherlands.
³ Department of Cell and Chemical Biology, The Institute for Chemical Immunology, Leiden University Medical Center LUMC, Leiden, The Netherlands.
⁴ Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center LUMC, Leiden, The Netherlands.
⁵ Institute of Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, Barcelona, Spain; Department of Biomedical Engineering, Institute of Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: [email protected].
⁶ Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Leiden, The Netherlands. Electronic address: [email protected].
⁷ Institute of Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, Barcelona, Spain; Department of Electronics and Biomedical Engineering, Faculty of Physics, Universitat de Barcelona, Barcelona, Spain. Electronic address: [email protected].

PMID: 33707017
DOI: 10.1016/bs.mcb.2020.09.001

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.

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Microscopy, Electron
Microscopy, Electron, Transmission
Microscopy, Fluorescence
Single Molecule Imaging*
Staining and Labeling