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pan-ASLM: Axially Swept Light Sheet Microscopy for Fast and High-Resolution Imaging of Expanded Samples.

Hannahmariam T Mekbib1,2, Lasse Pærgård Andersen2,3, Shuwen Zhang2

  • 1Department of Biomedical Engineering, Yale University, New Haven, CT, USA.

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Summary
This summary is machine-generated.

We developed pan-ASLM, a novel microscope for expansion microscopy. This advanced light-sheet microscope enables high-resolution imaging of large biological sample volumes, revealing nanoscale cellular details.

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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Expansion microscopy (ExM) significantly enhances resolution but requires advanced imaging systems.
  • Standard confocal microscopes struggle to image large, expanded samples efficiently.
  • High-resolution, large field-of-view microscopy is crucial for ExM applications.

Purpose of the Study:

  • To develop and validate a novel microscope optimized for imaging expansion microscopy samples.
  • To overcome the limitations of conventional microscopes in imaging large, high-resolution ExM datasets.
  • To introduce the pan-ASLM, a light-sheet microscope designed for expanded biological specimens.

Main Methods:

  • Development of pan-ASLM, a light-sheet microscope based on Axially Swept Light Sheet Microscopy (ASLM).
  • Utilizing a large field of view (640 x 640 µm²) and high resolution (586 nm lateral, 428 nm axial).
  • Imaging expanded HeLa cells and mouse kidney and brain tissues.

Main Results:

  • pan-ASLM achieves high resolution and a large field of view suitable for expanded samples.
  • Demonstrated imaging speed of up to 183 Mvoxels/sec (20 fps).
  • Achieved ~1200x higher imaging speed, ~7x larger field of view, and ~2x better axial resolution compared to standard confocal microscopy.

Conclusions:

  • pan-ASLM significantly advances imaging capabilities for expansion microscopy.
  • The microscope enables detailed nanoscale imaging of large biological volumes.
  • Validated for imaging both cell cultures and tissue samples, showcasing broad applicability.