Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy.

Area of Science:

• Biophysics
• Developmental Biology
• Microscopy

Background:

• Live imaging of large biological specimens is limited by light microscopes' shallow optical penetration.
• Simultaneous multi-view recording is crucial for comprehensive imaging of extensive biological samples.

Purpose of the Study:

• To develop a novel technology framework, SiMView, for high-speed, in vivo imaging of large biological specimens.
• To overcome the limitations of optical penetration depth in light microscopy for extensive sample coverage.

Main Methods:

• Development of SiMView: a light-sheet microscope with four synchronized optical arms.
• Real-time electronics for high-speed (175 million voxels/sec) sCMOS-based image acquisition.
• Computational modules for image registration, segmentation, tracking, and data management of terabyte-scale datasets.

Main Results:

• Successful implementation of one-photon and multiphoton SiMView systems.
• Acquisition of cellular dynamics in entire Drosophila melanogaster embryos at 30-second temporal resolution throughout development.
• High-resolution, long-term imaging of developing nervous systems and in vivo neuroblast cell lineage tracking.

Conclusions:

• SiMView provides quantitative morphological data for fast biological processes.
• Enables accurate, automated cell tracking across entire early embryos.
• Significantly advances in vivo imaging capabilities for large biological specimens.