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Related Experiment Video

Updated: Jun 6, 2025

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion STED Microscopy

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion STED Microscopy

Published on: April 9, 2018

Super-Resolution Imaging of Mitotic Spindle Microtubules Using STED Microscopy.

Isabella Koprivec¹, Valentina Štimac², Iva M Tolić³

¹Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia. Isabella.Koprivec@irb.hr.

Methods in Molecular Biology (Clifton, N.J.)

|December 1, 2024

Summary

Stimulated emission depletion (STED) microscopy offers super-resolution insights into cell division. This study details STED methods for analyzing microtubules and their attachments during mitosis in human cells.

Keywords:

Attachments Cell division Chromosomes Kinetochores Microtubules Mitosis Mitotic spindle Nucleation STED microscopy Segregation errors

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Last Updated: Jun 6, 2025

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Area of Science:

Cell Biology
Microscopy
Molecular Biology

Background:

Stimulated emission depletion (STED) microscopy is a super-resolution technique.
Mitosis research has recently benefited from STED microscopy.
STED microscopy is valuable for studying microtubules, kinetochore-microtubule attachments, and chromosome segregation errors.

Purpose of the Study:

To describe immunofluorescence combined with STED microscopy for analyzing microtubules and kinetochore-microtubule attachments in human mitotic spindles.
To describe live-cell STED microscopy for imaging transient processes in mitotic spindles.
To outline image analysis for quantifying microtubule bundles within the spindle.

Main Methods:

Immunofluorescence combined with STED microscopy.
Live-cell STED microscopy for short-term imaging.
Image analysis for quantitative assessment of microtubule bundles.

Main Results:

STED microscopy enables detailed analysis of microtubules and kinetochore-microtubule attachments in human mitotic spindles.
Live-cell STED microscopy allows imaging of dynamic events in crowded spindle regions.
Quantitative image analysis methods are established for microtubule bundles.

Conclusions:

STED microscopy is a powerful tool for investigating mitosis.
The described methods facilitate the study of microtubule dynamics and chromosome segregation.
This work provides a framework for advanced analysis of the mitotic spindle.