Dual-channel event microscopy for ultrafast biological imaging.

Area of Science:

• Biomedical Imaging
• Microscopy
• Optical Engineering

Background:

• Fast biological processes require advanced imaging techniques.
• Existing microscopy methods face trade-offs in speed, spectral capacity, and depth of field.
• Visualizing dynamic multicellular interactions in 3D tissue is challenging.

Purpose of the Study:

• To develop a novel microscopy technique for ultrafast, dual-channel imaging.
• To overcome limitations of current imaging modalities in speed, spectral capacity, and depth of field.
• To enable visualization of rapid biological dynamics in living systems.

Main Methods:

• Integration of digital micromirror device-based pulsed illumination, extended-depth-of-field (EDOF) optics, and event-based sensing.
• Development of Dual-Channel Event Microscopy (DEM).
• Utilized dual-color fluorescent phantoms and microsphere flow assays for validation.

Main Results:

• DEM achieves ultrafast, dual-channel EDOF imaging over a large field of view with significant depth of field.
• Accurate spectral separation and reconstruction of rapid motion at kilohertz frame rates were demonstrated.
• In vivo imaging visualized neutrophils and tumors in zebrafish, and revealed fine vascular networks and cardiac circulation.

Conclusions:

• DEM provides a powerful platform for visualizing rapid multicellular interactions and physiological dynamics.
• The technology enables quantitative mapping of blood-flow dynamics with high temporal fidelity.
• DEM overcomes existing trade-offs, offering simultaneous high temporal resolution, multicolor capability, and large axial coverage.