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Updated: Jun 15, 2025

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One step 4x and 12x 3D-ExM: robust super-resolution microscopy in cell biology.

Roshan X Norman1,2,3, Yu-Chia Chen2,4, Emma E Recchia2

  • 1Biophysics Graduate Program, University of Wisconsin-Madison, Madison, Wisconsin.

Biorxiv : the Preprint Server for Biology
|August 26, 2024
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Expansion Microscopy (ExM) offers advanced super-resolution imaging for cell biology. Optimized 4x and 12x 3D-ExM methods provide cost-effective, user-friendly nanoscale visualization of cellular structures.

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Area of Science:

  • Cell Biology
  • Microscopy
  • Biophysics

Background:

  • Super-resolution microscopy provides nanometer-scale resolution for biological imaging.
  • Expansion Microscopy (ExM) offers advantages over traditional methods like electron microscopy.
  • ExM enables simultaneous labeling and simpler sample preparation.

Purpose of the Study:

  • Introduce optimized 4-fold and 12-fold 3D-isotropic and preserved Expansion Microscopy (4x and 12x 3D-ExM) methods.
  • Demonstrate the utility of 3D-ExM for high-resolution imaging in cell biology research.

Main Methods:

  • Developed and optimized 4x and 12x 3D-ExM protocols.
  • Utilized standard confocal microscopy for imaging expanded samples.
  • Applied methods to 2D and 3D cell culture models.

Main Results:

  • Achieved robust and reproducible 3D isotropic expansion.
  • 12x 3D-ExM yielded a lateral resolution under 30 nm with standard confocal microscopy.
  • Enabled visualization of nanoscale structures like chromosomes, kinetochores, and viral particles.

Conclusions:

  • 3D-ExM is a cost-effective and user-friendly super-resolution technique.
  • Optimized 3D-ExM methods are suitable for diverse cell biology applications.
  • Facilitates detailed studies of cellular and chromatin architectures at the nanoscale.