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Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy f3D-SIM
Published on: September 29, 2014
Studying the Spatial Organization of ESCRTs in Cytokinetic Abscission Using the High-Resolution Imaging Techniques
Shai Adar-Levor1, Inna Goliand1, Michael Elbaum2
1Department of Life Sciences and NIBN, Ben Gurion University of the Negev, Beer Sheva, Israel.
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.
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