Axially swept dithered light-sheet microscope to reveal cardiac morphology

Area of Science:

• Biomedical Engineering
• Optical Imaging
• Cardiovascular Research

Background:

• Assessing cardiovascular development and disease requires understanding cardiac microstructure and vascular networks.
• Light-sheet microscopy with tissue clearing offers high-resolution organ imaging but struggles with complex myocardial structures.
• Existing methods face limitations in resolution, imaging depth, and speed for trabeculated tissues.

Purpose of the Study:

• To overcome limitations in light-sheet microscopy for imaging trabeculated myocardium.
• To develop a method for uniform, high-resolution volumetric imaging of intact cardiovascular systems.
• To enhance imaging throughput and preserve resolution across large biological specimens.

Main Methods:

• Implemented high-speed dithered light-sheet (DiLS) illumination to extend the confocal region and improve the space-bandwidth product.
• Integrated DiLS with an axial sweeping approach to create axially swept dithered light-sheet (AS-DiLS) microscopy.
• Utilized temporal dynamics for illumination-detection interplay to maintain uniform resolution.

Main Results:

• AS-DiLS extended the effective confocal region by over 40% while preserving optical sectioning.
• Achieved uniform illumination over a 12.5-millimeter range with enhanced imaging throughput.
• Delivered near-isotropic resolution (~2.5 μm) suitable for intricate cardiac structures.

Conclusions:

• AS-DiLS microscopy provides a scalable platform for comprehensive cardiovascular morphology and topology assessment.
• The technique enables detailed investigation of ventricular trabeculae, vasculature, and extracellular matrix.
• AS-DiLS is applicable for imaging cardiovascular systems across developmental stages, from embryos to adults.