High numerical aperture confocal volumetric mesoscope reveals mesoscale subcellular dynamics in vivo.
basic_science · Level V
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- Record sourced from PubMed, PMID 42458054.
- Also identified by DOI 10.1038/s41587-026-03231-z.
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Abstract
Simultaneously capturing subcellular dynamics across thousands of cells in mammals remains challenging because of trade-offs among spatial resolution, field of view, imaging speed and sample viability, particularly under high-NA (numerical aperture) conditions. Here we design a high-NA mesoscale objective lens and present a confocal volumetric mesoscope with an NA of 1.05, RUSH3D-HR, achieving ~390-nm resolution across a volume of 2.7 × 2.0 × 0.04 mm<sup>3</sup> at five volumes per second with low phototoxicity. By integrating our previously developed confocal scanning light-field framework with a parallelized deep-learning pipeline, RUSH3D-HR increases experimental efficiency by orders of magnitude. Long-term imaging at 70 billion voxels per second visualizes immune responses such as long-distance collective cell migrations during wound healing and large-scale migrasome generation and neutrophil-macrophage interactions during acute liver failure. Hour-long monitoring over 6,000 neutrophils reveals transient emergent swarming behaviors in mouse spleen during lipopolysaccharide-induced inflammation, which is suppressed with electroacupuncture at ST36, providing direct evidence for the immunomodulatory mechanism of peripheral electrical stimulation.