Versatile high-speed volumetric imaging from microscopic to macroscopic scale by self-adaptive oblique plane...

Area of Science:

• Advanced microscopy techniques
• Biomedical imaging
• Neuroscience research

Background:

• Increasing demand for high-speed, large-scale volumetric recording in complex multi-cellular systems.
• Oblique plane microscopy (OPM) offers real-time 4D microscopy but faces limitations in scaling beyond 1mm.
• Current OPM methods struggle with large specimens like the living mouse brain.

Purpose of the Study:

• To develop a self-adaptive OPM system overcoming field-of-view and resolution limitations.
• To enable flexible, high-speed volumetric imaging across diverse biological scales.
• To facilitate advanced imaging of dynamic processes in large multi-cellular models.

Main Methods:

• Leveraging Abbe's sine condition for a self-adaptive OPM.
• Achieving switchable imaging resolution, volume, and speed.
• Implementing rapid optical scanning and remote focusing for real-time 4D data acquisition.

Main Results:

• Demonstrated flexibility across a range of fields-of-view (up to 8 mm²) and resolutions (down to 2.2 µm³).
• Enabled brain-wide single neuron volumetric calcium imaging in zebrafish (1×0.4 mm² FOV at 5 Hz).
• Achieved capillary blood cell tracking in mouse brain (>3×3 mm² FOV) at 100 Hz.
• Showcased screen-and-zoom capability for subcellular structure visualization in cleared mouse brains.

Conclusions:

• The self-adaptive OPM provides a versatile platform for studying dynamic processes in multi-cellular systems.
• This technology supports both in vivo and fixed, optically cleared samples.
• Enables unprecedented large-scale, high-speed volumetric imaging for biological discovery.