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Related Concept Videos

Studying the Cytoskeleton01:17

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
Attachment of Sister Chromatids02:57

Attachment of Sister Chromatids

As cells progress into mitosis, the nuclear envelope breaks down, and the condensed chromosomes are exposed to the array of bipolar microtubules of the mitotic spindle. The kinetochore, a large, disc-shaped protein complex, is present at the centromere region of the sister chromatids and acts as a binding site for the microtubules.  Usually, the plus-end of a single microtubule is embedded within the kinetochore. However, some kinetochores first establish lateral contact with the side-wall of a...

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Related Experiment Video

Updated: May 8, 2026

Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy
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Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy

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Studying kinetochore-fiber ultrastructure using correlative light-electron microscopy.

Daniel G Booth1,2, Liam P Cheeseman1, Ian A Prior1

  • 1Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool L69 3BX, U.K.

Methods in Cell Biology
|August 27, 2013
PubMed
Summary

Correlative light-electron microscopy (CLEM) enables researchers to study cellular structures like the mitotic spindle by combining light microscopy and electron microscopy. This guide simplifies CLEM for detailed ultrastructural analysis of cell division.

Keywords:
Correlative electron microscopyKinetochore-fiberMicrotubuleMitosisMitotic spindle

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Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Electron microscopy (EM) provides high-resolution imaging of intracellular structures.
  • Correlative light-electron microscopy (CLEM) bridges light microscopy (LM) and EM for targeted ultrastructural analysis.
  • Studying the mitotic spindle requires high resolution to visualize microtubules and associated structures.

Purpose of the Study:

  • To present CLEM as an effective method for studying the mitotic spindle apparatus.
  • To provide a detailed, step-by-step guide for achieving successful CLEM.
  • To demonstrate how varying sectioning planes enhances spindle and microtubule ultrastructural analysis.

Main Methods:

  • Application of CLEM to visualize cells during mitosis.
  • Step-by-step pictorial guidance for CLEM sample preparation and imaging.
  • Analysis of mitotic spindles and microtubules using varied sectioning planes in CLEM.

Main Results:

  • Successful application of CLEM to track specific mitotic cells from LM to EM.
  • Demonstration of how CLEM facilitates detailed ultrastructural examination of the spindle apparatus.
  • Insights into kinetochore fiber ultrastructure through multi-angle analysis enabled by CLEM.

Conclusions:

  • CLEM is a valuable and achievable technique for detailed study of the mitotic spindle.
  • The provided guide simplifies CLEM, making it accessible for researchers.
  • CLEM, with adjustable sectioning, offers novel perspectives on microtubule and kinetochore fiber ultrastructure.