Capturing Endocytic Segregation Events with HPF-CLEM

Edward Brown, Jan Van Weering, Thom Sharp, Judith Mantell, Paul Verkade

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

18 Citations (Scopus)

Abstract

We have advocated the use of high-pressure freezing (HPF) in specific types of Correlative Light Electron Microscopy (CLEM) experiments because the intracellular components such as the cytoskeleton and membrane tubules can only be adequately preserved via cryofixation. To allow fast transfer from the light microscope into a cryofixation device, we have developed the Rapid Transfer System (RTS) for the EMPACT2 high-pressure freezer. In this chapter, we will describe how to prepare and perform a CLEM experiment using this device and will highlight the latest changes made to the original system to optimize the workflow.

Original languageEnglish
Title of host publicationMethods in Cell Biology
PublisherAcademic Press Inc.
Pages175-201
Number of pages27
DOIs
Publication statusPublished - 2012

Publication series

NameMethods in Cell Biology
Volume111

Keywords

  • Correlative microscopy
  • Electron microscopy
  • Electron tomography.
  • Freeze substitution
  • High-pressure freezing
  • Live-cell imaging
  • Probes

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