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Updated: Jan 13, 2026

Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi
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Unraveling Subcellular Ultrastructure with Cyclically Multiplexed Expansion Microscopy.

Seweryn Gałecki1,2,3, Bo-Jui Chang1,2, Felix Zhou1,2

  • 1Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, TX 75390, USA.

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|January 9, 2026
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Summary
This summary is machine-generated.

We developed Cy-ExM, a novel high-plex imaging strategy, to overcome limitations in dense cellular ultrastructure mapping. This method enables high-resolution, 3D imaging of 20 targets within entire mammalian cells.

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Area of Science:

  • Cellular Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Fluorescence microscopy faces challenges like spectral overlap and limited resolution, hindering detailed cellular ultrastructure mapping.
  • Dense mapping of cellular components is crucial for understanding cell biology and disease mechanisms.

Purpose of the Study:

  • To develop a high-plex imaging strategy overcoming current fluorescence microscopy limitations.
  • To enable dense, 3D super-resolution mapping of the full cellular volume.

Main Methods:

  • Cy-ExM integrates optimized cryo-fixation for antigen preservation.
  • Expansion microscopy is combined with iterative immunofluorescence labeling.
  • Oblique plane microscopy is utilized for 3D super-resolution imaging.

Main Results:

  • Successfully imaged 20 biological targets within individual mammalian cells.
  • Achieved high-plex, 3D super-resolution imaging of the complete cellular volume.
  • Demonstrated Cy-ExM's capability to overcome spectral overlap and resolution limitations.

Conclusions:

  • Cy-ExM is a powerful high-plex imaging strategy for detailed cellular ultrastructure analysis.
  • This approach advances the dense mapping of biological targets in 3D.
  • Enables deeper insights into cellular organization and function.