A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues

Area of Science:

• Biomedical Engineering
• Microscopy
• Neuroscience

Background:

• Light-sheet microscopy is crucial for high-throughput volumetric imaging of cleared tissues.
• Existing systems lack flexibility for varied imaging requirements (resolution, sample size, clearing methods, sample holders).

Purpose of the Study:

• To present a hybrid open-top light-sheet (OTLS) microscopy system.
• To demonstrate its versatility for multi-scale imaging of cleared tissues with diverse requirements.

Main Methods:

• Development of a hybrid OTLS system combining non-orthogonal dual-objective and conventional orthogonal architectures.
• Application of the system for imaging cleared mouse brains and multiple specimens.
• Demonstration of sub-micrometer resolution imaging of sparse axons and high-throughput automated imaging.

Main Results:

• Efficient screening and targeted sub-micrometer imaging of sparse axons in a cleared mouse brain.
• High-throughput automated imaging of multiple specimens, including quantitative multi-scale analysis of brain metastases.
• The hybrid OTLS system shows superior versatility and performance compared to existing systems.

Conclusions:

• The hybrid OTLS system provides a versatile and high-performance solution for cleared-tissue imaging.
• It addresses the diverse and growing needs of researchers in various biological applications.