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Light-sheet Fluorescence Microscopy for the Study of the Murine Heart

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Published on: September 15, 2018

Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution.

Tonmoy Chakraborty^1,2, Meghan K Driscoll², Elise Jeffery³

¹Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.

|November 2, 2019

Summary

Cleared-tissue axially swept light-sheet microscopy (ctASLM) provides high-resolution 3D imaging of cleared tissues. This advanced light-sheet microscopy technique enables subcellular resolution for detailed biological analysis.

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Area of Science:

Biomedical Imaging
Microscopy Techniques
Cell Biology

Background:

High-resolution imaging of cleared tissues is crucial for understanding complex biological structures.
Existing light-sheet microscopy methods face limitations in resolution, field of view, and compatibility with various clearing media.

Purpose of the Study:

To introduce and validate cleared-tissue axially swept light-sheet microscopy (ctASLM) for advanced tissue imaging.
To demonstrate ctASLM's capability for isotropic, subcellular resolution imaging in diverse cleared tissue preparations.

Main Methods:

Development of axially swept light-sheet microscopy optimized for cleared tissues.
Imaging of millimeter-scale cleared tissues using aqueous and non-aqueous immersion media.
Utilizing high optical sectioning and a large field of view for comprehensive tissue visualization.

Main Results:

Achieved up to 260 nm axial resolution, a significant improvement over conventional methods.
Enabled 3D imaging of cleared tissues with subcellular resolution.
Facilitated automated detection of multicellular architectures, individual cells, and synaptic structures.

Conclusions:

ctASLM offers a powerful new tool for high-resolution imaging of cleared biological tissues.
The technique supports imaging of live, expanded, and chemically cleared samples.
ctASLM advances the study of cellular and subcellular details within large tissue volumes.