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Author Spotlight: Customized Light-Sheet Imaging for Investigating Myocardial Structures in Rodent Hearts

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A Versatile Tiling Light Sheet Microscope for Imaging of Cleared Tissues.

Yanlu Chen¹, Xiaoliang Li², Dongdong Zhang³

¹Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang 310024, China; Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang 310024, China.

|November 4, 2020

Summary

This summary is machine-generated.

This study introduces a novel tiling light sheet microscope for rapid, high-resolution 3D imaging of cleared tissues. Its phase modulation technology enhances accuracy and ease of use for multicolor imaging applications.

Keywords:

cleared tissue imaging light sheet microscopy tiling light sheet microscopy tiling light sheet selective plane illumination microscopy tissue clearing tissue expansion

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Area of Science:

Microscopy
Biotechnology
Optical Imaging

Background:

Conventional light sheet microscopy faces limitations in spatial resolution and optical sectioning for cleared tissues.
Achieving high-resolution 3D imaging of intact biological samples requires advanced microscopy techniques compatible with tissue clearing.

Purpose of the Study:

To develop and present a tiling light sheet microscope optimized for rapid, multicolor 3D imaging of cleared tissues.
To achieve micron-scale to submicron-scale spatial resolution, with further enhancement via tissue expansion.

Main Methods:

Utilized tiling light sheets to improve spatial resolution and optical sectioning compared to conventional methods.
Employed phase modulation of illumination light to control light sheet properties and ensure microscope alignment.
Demonstrated compatibility with all tissue clearing methods for versatile application.

Main Results:

Achieved micron-scale (4 × 4 × 10 μm³) to submicron-scale (0.3 × 0.3 × 1 μm³) spatial resolution.
Enhanced resolving ability to sub-100 nm (70 × 70 × 200 nm³) through tissue expansion.
Validated the microscope's accuracy, reliability, and ease of operation for multicolor 3D imaging.

Conclusions:

The developed tiling light sheet microscope offers superior performance for imaging cleared tissues.
Phase modulation provides a robust mechanism for alignment and adaptability, enhancing imaging quality.
The microscope is a versatile tool for advanced 3D biological imaging research.