Studying the Spatial Organization of ESCRTs in Cytokinetic Abscission Using the High-Resolution Imaging Techniques SIM and Cryo-SXT

Shai Adar-Levor, Inna Goliand, Michael Elbaum, Natalie Elia

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

Abstract

The ESCRT machinery mediates scission of the intercellular bridge that connects two daughter cells at the end of cytokinesis. Structured illumination microscopy (SIM) and cryo-soft-X-ray tomography (cryo-SXT) have been used in recent years to study the topology of ESCRT-driven cytokinetic abscission. These studies revealed that the intercellular bridge is occupied by cortical rings and spiral-like filaments and that ESCRTs form ring-like structures in this region during abscission. In this chapter, we provide two protocols: a protocol for determining the spatial organization of specific ESCRT components at the intercellular bridge using SIM and a protocol for resolving the ultrastructural organization of cortical filaments at the intercellular bridge using cryo-SXT.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages129-148
Number of pages20
DOIs
StatePublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
Volume1998
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell division
  • Correlative imaging
  • Cryo
  • Cytokinesis
  • ESCRT
  • ESCRT-III
  • Fluorescence imaging
  • Immunostaining
  • Membrane fission
  • Soft-X-ray
  • Super-resolution microscopy
  • Synchronization
  • Tomography

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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