Hard-wired lattice light-sheet microscopy for imaging of expanded samples.

Area of Science:

• Neuroscience
• Biophysics
• Optical Microscopy

Background:

• Light-sheet fluorescence microscopy (LSFM) is crucial for 3D imaging of cells and tissues.
• Lattice light-sheet microscopy enhances axial resolution and imaging speed using Bessel beams.
• Conventional LSFM often uses Gaussian beams, limiting resolution and field size.

Purpose of the Study:

• To develop a facile and stable lattice light-sheet microscope.
• To achieve high-resolution, homogeneous imaging of expanded biological samples.
• To demonstrate the microscope's capability for dual-color imaging.

Main Methods:

• Fabrication of a fixed ring mask for lattice light-sheet generation.
• Utilized micro-optics for a stable illumination path, avoiding spatial light modulators.
• Employed long-working distance objectives for imaging optically cleared mouse hippocampus tissue.

Main Results:

• Achieved a homogeneous axial resolution of 1.2 µm.
• Demonstrated imaging over a field of view of (333 µm)².
• Showcased high-resolution imaging of expanded neurons and neurites.

Conclusions:

• The developed lattice light-sheet microscope offers a simple, stable, and effective platform for high-resolution biological imaging.
• This technology is well-suited for neuroscience applications, particularly imaging large, cleared tissue samples.
• The microscope provides excellent homogeneous resolution, enabling detailed investigation of cellular structures.