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Light-sheet Microscopy for Three-dimensional Visualization of Human Immune Cells
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Expansion and Light-Sheet Microscopy for Nanoscale 3D Imaging.

Luca Pesce1, Pietro Ricci2, Giancarlo Sportelli1

  • 1Department of Physics - Enrico Fermi, University of Pisa, Largo Pontecorvo, 3, Pisa, 56127, Italy.

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Summary

Expansion Microscopy (ExM) and Light-Sheet Fluorescence Microscopy (LSFM) combine for ExLSFM, offering enhanced nanoscale imaging of large biological samples. This integrated approach improves resolution and speed for advanced visualization.

Keywords:
3D reconstructionexpansion microscopylight‐sheet fluorescence microscopymultiplexed imagingnanoscale imaging

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Expansion Microscopy (ExM) enables nanoscale imaging by physically expanding samples.
  • Light-Sheet Fluorescence Microscopy (LSFM) provides rapid, high-resolution, volumetric imaging.
  • Integrating ExM and LSFM (ExLSFM) offers synergistic benefits for biological specimen visualization.

Purpose of the Study:

  • To review the combined advancements of ExM and LSFM, termed ExLSFM.
  • To highlight the performance and applications of the integrated ExLSFM technique.
  • To encourage wider adoption and further development of ExLSFM.

Main Methods:

  • Detailed review of chemical processes critical for ExM.
  • Analysis of tailored LSFM optical setups for expanded samples.
  • Emphasis on sample preparation adjustments for ExLSFM data acquisition.

Main Results:

  • ExLSFM demonstrates excellent performance by combining ExM and LSFM.
  • The review covers diverse specimen types successfully studied with ExLSFM.
  • Key chemical, optical, and sample preparation factors for ExLSFM are identified.

Conclusions:

  • ExLSFM represents a powerful integrated imaging modality.
  • Further development and application of ExLSFM are anticipated.
  • This review aims to foster greater understanding and innovation in ExLSFM.