Two-photon 3D imaging of optically stimulated neural activity at 100 Hz

Le résumé fourni par la source

Understanding how neurons integrate synaptic inputs requires imaging techniques capable of capturing rapid, three-dimensional dendritic events. These processes occur on millisecond timescales and submicron spatial scales, exceeding the speed of conventional two-photon microscopy (2PM). We developed dual-view Bessel two-photon projection microscopy (dv-B2PM), a high-speed volumetric imaging approach that achieves 100 Hz whole-volume acquisition with synaptic-level resolution. dv-B2PM simultaneously records two orthogonal projections of the same 3D volume, preserving spatial information while minimizing ambiguity from structural overlap. Combining dv-B2PM with two-photon glutamate uncaging, we visualized 3D Ca²⁺ dynamics in neurons following localized stimulation. Multi-timescale analysis revealed dendrite-to-soma Ca²⁺ signal propagation, back propagated Ca²⁺ signal from the soma, and multi-frequency (5-40 Hz) Ca²⁺ transients activated along apical dendrites at speeds from ten of microns per second to millimeters per seconds. These findings demonstrate dv-B2PM as a powerful tool for direct visualization of 3D calcium dynamics associated with dendritic integration across extended neuronal structures, bridging the gap between optical imaging and the dynamic biophysics of neuronal integration.

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