Correlative light-electron microscopy as a tool to study in vivo dynamics and ultrastructure of intracellular structures

Elena V. Polishchuk, Roman S. Polishchuk, Alberto Luini

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Correlative light-electron microscopy (CLEM) is a very effective technique that combines live-cell imaging and immuno-electron microscopy for ultrastructural morphological characterization of dynamic intracellular organelles. The use of green fluorescent protein (GFP)-tagged chimeras allows the user to follow the movements and/or behavior of intracellular structures in a live cell and to fix it at the moment of interest. The subsequent immuno-electron microscopy processing can then reveal the three-dimensional architecture of the same structure, together with precise recognition of the GFP-labeled protein. The process resembles the taking of a high-resolution snapshot of an interesting live scene. Considering that CLEM is a very useful but technically demanding and time-consuming technique, accurate protocols will be helpful to simplify the work of scientists who are willing to apply this method for their own purposes. Here, we present a detailed protocol that describes all of the "tricks" and know-hows involved in carrying out the crucial steps of a CLEM experiment

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages413-422
Number of pages10
Volume931
ISBN (Print)9781627030557
DOIs
Publication statusPublished - 2012

Publication series

NameMethods in Molecular Biology
Volume931
ISSN (Print)10643745

Keywords

  • Correlative light-electron microscopy
  • Gold enhancement
  • Green fluorescent protein
  • Gridded coverslips
  • Immuno-electron microscopy
  • Live-cell imaging
  • Paraformaldehyde-glutaraldehyde fixation
  • Resin embedding

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Polishchuk, E. V., Polishchuk, R. S., & Luini, A. (2012). Correlative light-electron microscopy as a tool to study in vivo dynamics and ultrastructure of intracellular structures. In Methods in Molecular Biology (Vol. 931, pp. 413-422). (Methods in Molecular Biology; Vol. 931). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-56-4_20