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

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 publicationMethods in Cell Biology
PublisherAcademic Press Inc.
DOIs
Publication statusAccepted/In press - 16 Oct 2020

Publication series

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

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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