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Measuring abscission spatiotemporal dynamics using quantitative high-resolution microscopy.

O Gershony1, S Sherman1, S Adar1

  • 1Ben Gurion University of the Negev, Beer Sheva, Israel.

Methods in Cell Biology
|January 10, 2017
PubMed
Summary
This summary is machine-generated.

Researchers quantitatively studied Endosomal Sorting Complex Required for Transport (ESCRT)-mediated cell division using advanced microscopy. This work details methods to analyze ESCRT protein dynamics and structure during cell abscission.

Keywords:
Cell cycleCell divisionCytokinesisESCRTIntercellular bridgeLive cell imagingMembrane fissionStructured illumination microscopy

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Microscopy

Background:

  • Endosomal Sorting Complex Required for Transport (ESCRT) machinery mediates crucial cellular processes, including cell division.
  • Cytokinetic abscission, the final step in cell division, is a complex process involving ESCRT proteins.
  • Understanding the spatiotemporal dynamics of ESCRT during abscission is vital for cell biology.

Purpose of the Study:

  • To describe the application of spinning disk live cell imaging and structured illumination microscopy (SIM) for analyzing ESCRT-mediated abscission.
  • To quantitatively define the dynamics and structural organization of abscission.
  • To correlate structural data from SIM with dynamic information from live cell imaging.

Main Methods:

  • Quantitative live cell imaging using spinning disk microscopy.
  • High-resolution imaging with structured illumination microscopy (SIM).
  • Development of a protocol to integrate dynamic and structural data.

Main Results:

  • The study provides a detailed methodology for observing ESCRT-mediated abscission.
  • It enables quantitative analysis of protein dynamics and structural organization during the process.
  • A protocol is presented to correlate structural and dynamic data.

Conclusions:

  • Spinning disk microscopy and SIM are powerful tools for studying the spatiotemporal characteristics of ESCRT-mediated abscission.
  • The presented protocol allows for a comprehensive quantitative analysis of this critical cell division event.
  • This approach enhances our understanding of the molecular mechanisms governing cell abscission.