Isotropic, aberration-corrected light sheet microscopy for rapid high-resolution imaging of cleared tissue

Area of Science:

• Biomedical Imaging
• Optical Microscopy
• Biotechnology

Background:

• Light-sheet microscopy enables large-scale tissue imaging.
• Current limitations include slow imaging speeds and aberrations, hindering high-resolution analysis of centimeter-sized samples.

Purpose of the Study:

• To develop a compact, high-speed light-sheet fluorescence microscope.
• To achieve high isotropic resolution (850 nm) across large cleared tissues (1 cm³).
• To overcome limitations of speed and optical aberrations in existing systems.

Main Methods:

• Utilized off-the-shelf optics including an air objective and meniscus lens.
• Implemented an axially swept light sheet for diffraction-limited resolution and aberration correction.
• Employed a concave mirror for field curvature correction and a closed-loop feedback system to enhance imaging speed.

Main Results:

• Achieved 850 nm isotropic resolution in cleared samples up to 1 cm³ across various refractive indices (1.33-1.56).
• Enhanced imaging speed tenfold to 100 frames per second.
• Demonstrated system performance from subcellular structures to centimeter scales.

Conclusions:

• The developed microscope offers a significant advancement for high-resolution, large-scale tissue imaging.
• The system effectively corrects aberrations and increases imaging speed, enabling faster and more detailed analysis of cleared biological samples.
• This technology is suitable for diverse applications in biological research requiring deep tissue visualization.