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Updated: Oct 3, 2025

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Visualizing cellular and tissue ultrastructure using Ten-fold Robust Expansion Microscopy (TREx).

Hugo G J Damstra1, Boaz Mohar2, Mark Eddison2

  • 1Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, Utrecht, Netherlands.

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|February 18, 2022
PubMed
Summary
This summary is machine-generated.

Ten-fold Robust Expansion Microscopy (TREx) expands biological samples tenfold, enabling high-resolution imaging of cells and tissues without specialized equipment. This method enhances subcellular detail for protein localization studies.

Keywords:
cell biologyexpansion microscopyhumanimmunofluorescencelight microscopymouseneurosciencesub-organelle imagingsuper-resolution

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Area of Science:

  • Microscopy and Imaging Technologies
  • Biotechnology and Bioengineering
  • Cell Biology and Histology

Background:

  • Expansion Microscopy (ExM) overcomes light microscopy's diffraction limit by physically expanding samples.
  • Original ExM offers a four-fold expansion, limiting ultimate resolution.
  • Existing advanced ExM methods often sacrifice ease of use or versatility.

Purpose of the Study:

  • To systematically explore the ExM recipe space.
  • To develop a novel, high-expansion ExM method with improved usability.
  • To enable high-resolution subcellular imaging in diverse biological samples.

Main Methods:

  • Developed Ten-fold Robust Expansion Microscopy (TREx), a novel ExM protocol.
  • TREx utilizes a swellable polymer gel for isotropic sample expansion.
  • Applied TREx to thick mouse brain sections and cultured human cells.

Main Results:

  • TREx gels achieve a ten-fold expansion factor.
  • The TREx method requires no specialized equipment or procedures.
  • High-resolution subcellular imaging was achieved in both tissue and cell samples.
  • TREx successfully combined antibody staining with small-molecule stains for ultrastructural context.

Conclusions:

  • TREx offers a versatile and user-friendly method for achieving ten-fold expansion in ExM.
  • This technique significantly enhances subcellular resolution for imaging applications.
  • TREx provides ultrastructural context for protein localization studies.