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Updated: May 20, 2025

Measurement of Microtubule Dynamics by Spinning Disk Microscopy in Monopolar Mitotic Spindles
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Microtubules as a versatile reference standard for expansion microscopy

Rajdeep Chowdhury1,2, Tiago Mimoso3, Abed Alrahman Chouaib4

  • 1Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.

Communications Biology
|March 27, 2025
PubMed

Insights

Microtubules are proposed as ideal structures for validating expansion microscopy (ExM) techniques. This study details methods for using microtubules at various scales to assess ExM resolution and precision.

Area of Science:

  • Biotechnology
  • Microscopy
  • Cell Biology

Background:

  • Expansion microscopy (ExM) is advancing, requiring standardized validation methods.
  • Current ExM validation structures lack consensus, hindering technique development.

Purpose of the Study:

  • To propose and validate microtubules as a universal standard for ExM validation.
  • To establish protocols for using microtubules across different ExM resolution scales.

Main Methods:

  • Immunostaining of cellular microtubules for validation of techniques with <50 nm resolution.
  • In vitro assembly and imaging of microtubules for validation of techniques with ~10 nm resolution.
  • Labeling tubulin peptide chains and analyzing single tubulin molecules for nanometer-scale techniques.

Main Results:

  • Microtubule diameter is suitable for validating ExM resolutions.
  • Protocols for in vitro microtubule assembly and cellular extraction are presented.
  • Multiple methods demonstrate microtubule utility for ExM validation.

Conclusions:

  • Microtubules offer a versatile and reliable structure for ExM validation.
  • The proposed methods support the standardization of ExM technology assessment.
  • Microtubules are valuable for validating ExM and related super-resolution imaging technologies.

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