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Related Concept Videos

Centrioles and Centrosomes01:13

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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...
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Related Experiment Video

Updated: May 5, 2026

Isolation and Fluorescence Imaging for Single-particle Reconstruction of Chlamydomonas Centrioles
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High-fold Homogeneous Expansion Microscopy Reveals Ultrastructural Centrioles.

Wen-Qing Yang1,2, Ting-Jui Ben Chang1,2, Liang-Chen Pan1,2

  • 1Department of Electrical Engineering, National Taiwan University, Taipei 10617, Taiwan.

ACS Nano
|May 4, 2026
PubMed
Summary

High-fold homogeneous expansion microscopy (hiHomoExM) achieves uniform ∼8-9x expansion, preserving cellular ultrastructure. This technique, combined with super-resolution microscopy, offers robust nanoscale imaging for advanced cellular structural analysis.

Keywords:
ExMcentriolesdSTORMexpansionhigh-foldsuper-resolutionultrastructure

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Area of Science:

  • Cell Biology
  • Microscopy
  • Biophysics

Background:

  • Super-resolution microscopy enables nanoscopic visualization of biomolecules.
  • Expansion microscopy (ExM) offers an optics-independent route to super-resolution by physically enlarging specimens.
  • Existing high-fold ExM methods often lack validation for preserving ultrastructural uniformity.

Purpose of the Study:

  • To introduce high-fold homogeneous expansion microscopy (hiHomoExM) for uniform, high-fold specimen expansion.
  • To validate hiHomoExM's ability to preserve cellular ultrastructure across diverse biological targets.
  • To enhance spatial resolution by integrating hiHomoExM with iterative expansion and super-resolution microscopy.

Main Methods:

  • Development of hiHomoExM, a single-round ExM technique for uniform ∼8-9x expansion.
  • Application of hiHomoExM to various cellular structures, including centrioles and nuclear pore complexes.
  • Integration of hiHomoExM with iterative expansion and direct STORM (dSTORM) for enhanced resolution (hiHomoEx-dSTORM).

Main Results:

  • hiHomoExM achieved uniform ∼8-9x expansion while preserving ultrastructure in centrioles, NPCs, and other organelles.
  • Streamlined sample preparation and postexpansion labeling supported high labeling density and structural integrity.
  • hiHomoEx-dSTORM resolved previously elusive centriole ultrastructures, revealing detailed organization of CEP44, CCDC77, and SAS6 with ∼2 nm resolution.

Conclusions:

  • hiHomoExM provides a robust and accessible method for uniform, high-fold expansion microscopy.
  • The combined hiHomoExM and hiHomoEx-dSTORM platform bridges physical and optical super-resolution techniques.
  • This approach significantly advances cellular structural analysis by enabling nanoscale characterization of complex biomolecular assemblies.