Dual-Channel Event Microscopy for Ultrafast Biological Imaging

Area of Science:

• Biophotonics and Imaging
• Cellular and Molecular Imaging
• Physiological Dynamics

Background:

• Fundamental biological processes involve fast, multiscale dynamics across diverse cell types in 3D tissues.
• Existing imaging modalities face limitations in balancing speed, spectral capacity, depth of field, and resolution.
• Simultaneously capturing high temporal resolution, multicolor capability, and volumetric coverage is crucial for studying these dynamics.

Purpose of the Study:

• To introduce Dual-Channel Event Microscopy (DEM), an imaging system designed to overcome tradeoffs in current technologies.
• To demonstrate DEM's capability for ultrafast, dual-channel volumetric imaging across large fields of view.
• To showcase DEM's application in visualizing rapid multicellular interactions and physiological dynamics in living systems.

Main Methods:

• Integration of digital micromirror device (DMD)-based pulsed illumination.
• Incorporation of extended depth-of-field (DOF) optics.
• Utilization of event-based sensing for ultrafast, dual-channel volumetric imaging over a 2.3 × 1.3 mm² FOV with a 200 μm DOF.

Main Results:

• DEM achieved accurate spectral separation and reconstruction of rapid motion at kilohertz frame rates using fluorescent phantoms and flow assays.
• In vivo imaging successfully visualized neutrophils and premalignant tumors in freely swimming zebrafish.
• In immobilized specimens, DEM provided optical sectioning to reveal fine vascular networks and cardiac blood flow; it also mapped blood-flow dynamics in zebrafish tails with high temporal fidelity.

Conclusions:

• DEM unites ultrafast acquisition, dual-channel capability, volumetric coverage, and intrinsic optical sectioning in an event-driven architecture.
• This technology offers a powerful platform for visualizing rapid multicellular interactions and physiological dynamics in living systems.
• DEM overcomes previous imaging limitations, enabling unprecedented insights into complex biological processes.