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Updated: Jan 1, 2026
12:20
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
Published on: March 15, 2014
14.8K
Ultrastructural Analysis of Microtubule Ends
J Richard McIntosh1, Eileen O'Toole2, Cynthia Page2
1Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO, USA. richard.mcintosh@colorado.edu.
Methods in Molecular Biology (Clifton, N.J.)
|December 28, 2019
Summary
Electron microscopy reveals microtubule end structures during growth and shrinkage. Rapid freezing and electron tomography provide high-resolution details of microtubule dynamics in vivo and in vitro.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Light microscopy offers limited resolution for microtubule structures.
- Previous studies used indirect methods to infer microtubule end dynamics.
Purpose of the Study:
- To describe electron microscopy methods for visualizing microtubule ends during dynamic processes.
- To detail sample preparation techniques for preserving labile microtubule structures.
Main Methods:
- Rapid freezing (vitrification) for sample preservation.
- Electron tomography for 3D structural analysis.
- Methods for both in vivo (cells) and in vitro studies.
Main Results:
- Detailed ultrastructural visualization of microtubule ends during growth and shortening.
- Successful application of rapid freezing and electron cryo-tomography.
- Quantification of microtubule structures through model analysis.
Conclusions:
- Electron microscopy, particularly cryo-electron tomography, is crucial for understanding microtubule dynamics.
- Optimized sample preparation is key to preserving the labile nature of dynamic microtubules.
- 3D structural information is essential for comprehending microtubule growth and shortening pathways.
Keywords:
DepolymerizationElectron cryo-tomographyElectron microscopyElectron tomographyFreeze-substitutionFrozen-hydratedMicrotubulePolymerizationProtofilamentThree-dimensionalMore Related Videos
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