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SOLEIL: single-objective lens inclined light sheet localization microscopy.

Shih-Te Hung1, Jelmer Cnossen1, Daniel Fan1

  • 1Delft Center for Systems and Control, Delft University of Technology, Delft, Netherlands.

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|July 5, 2022
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Summary
This summary is machine-generated.

We developed a new microscope, SOLEIL, for advanced single-molecule localization microscopy (SMLM) in thick samples. This user-friendly system improves imaging resolution by reducing background noise, making complex biological samples easier to study.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • High numerical aperture (NA) light sheet illumination enhances single-molecule localization microscopy (SMLM) resolution by reducing background fluorescence.
  • Current high-NA SMLM methods often require complex, custom setups, limiting their practical application.

Purpose of the Study:

  • To develop a user-friendly, adaptable light sheet microscopy system for 2D and 3D SMLM in thick samples.
  • To integrate oblique illumination with point spread function (PSF) engineering for enhanced SMLM.
  • To benchmark the developed system against existing microscopy techniques.

Main Methods:

  • Development of the Single-Objective Lens-Inclined Light Sheet microscope (SOLEIL).
  • Integration of oblique illumination and PSF engineering for dSTORM imaging.
  • Compatibility testing with standard laboratory equipment and sample holders.
  • Optimization of oblique angle and light sheet thickness for optical sectioning.
  • Benchmarking SOLEIL against widefield and HILO microscopy using biological samples.

Main Results:

  • SOLEIL enables 2D and 3D SMLM in thick samples using standard components.
  • In 15 μm thick Caco2-BBE cells, SOLEIL achieved a 374% higher intensity-to-background ratio (vs. widefield) and a 54% improvement in the estimated Cramér-Rao Lower Bound (CRLB).
  • Compared to HILO microscopy, SOLEIL demonstrated a 184% higher intensity-to-background ratio and a 20% improvement in the estimated CRLB.

Conclusions:

  • SOLEIL offers a simplified and versatile approach to high-resolution SMLM in complex biological samples.
  • The system provides significant improvements in image quality and localization precision over conventional methods.
  • SOLEIL enhances the usability of advanced SMLM techniques without requiring specialized infrastructure.