Low-cost multimodal light sheet microscopy for optically cleared tissues and living specimens.

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Optical Engineering

Background:

• Light sheet microscopy is increasingly popular for cellular and tissue imaging due to its speed, low phototoxicity, and high signal-to-noise ratio.
• A major limitation is optical aberrations, particularly defocusing from refractive index variations in cleared tissues.
• Existing methods struggle to maintain image quality across diverse sample types and resolutions.

Purpose of the Study:

• To develop an inexpensive and easily constructible light sheet microscopy system.
• To overcome optical aberrations caused by refractive index mismatch in cleared biological samples.
• To enhance modularity for imaging a wide range of sample sizes and resolutions.

Main Methods:

• Designed a light sheet microscope with optomechanically decoupled sample scanning and detection.
• Incorporated a swappable excitation arm for versatile imaging capabilities.
• Tested the system on iDISCO cleared embryos and sciatic nerves.

Main Results:

• Successfully corrected optical aberrations, enabling clear imaging of cleared tissues.
• Achieved high-resolution 3D reconstruction of embryos and nerves within minutes.
• Demonstrated system's adaptability for imaging from single cells (µm resolution) to tissues (mm resolution).

Conclusions:

• The proposed light sheet microscopy system offers a cost-effective solution to imaging challenges in cleared biological samples.
• Its modular design and aberration correction capabilities expand the utility of light sheet microscopy for diverse research applications.
• This instrumentation facilitates rapid, high-fidelity 3D imaging across multiple biological scales.