Supercritical-lens light-sheet fluorescence microscopy with sub-diffraction-limit axial resolution, enhanced FOV, and...

Area of Science:

• Optics and Photonics
• Biomedical Imaging
• Microscopy Technology

Background:

• Light-sheet fluorescence microscopy (LSFM) is crucial for biological volumetric imaging.
• Current LSFM designs face limitations in axial resolution and field of view (FOV) due to bulky illumination paths and spatial constraints.
• Achieving higher numerical aperture (NA) often increases system complexity.

Purpose of the Study:

• To address the design challenges in LSFM for improved resolution and FOV.
• To introduce a novel supercritical lens (SCL) for generating a high-quality light sheet.
• To demonstrate a compact and high-performance LSFM system using SCL technology.

Main Methods:

• Designed a supercritical lens (SCL) to create a quasi-non-diffracting light sheet with suppressed sidelobes.
• Developed a light-sheet fluorescence microscope (LSFM) incorporating the SCL.
• Performed in-vivo volumetric imaging of zebrafish using the SCL-based LSFM.

Main Results:

• The SCL effectively suppressed sidelobes to 7.6% without a window.
• The SCL-based LSFM achieved sub-diffraction-limit axial resolution, exceeding theoretical NA limits.
• Enhanced FOV with minimal divergence was observed.
• The system demonstrated a simple, compact design using chip-scale components.

Conclusions:

• Supercritical lens technology offers a promising approach for advancing LSFM.
• The developed SCL-based LSFM enables high-performance 3D biological imaging.
• This work paves the way for future highly integrated, chip-based LSFM systems.