Ultrafast two-photon volumetric scanning microscopy

Here we achieved record-high >500 volumes/second two-photon imaging by improving lateral and axial scanning speed via 32-channel multifocal excitation/detection, and a tunable acoustic gradient-index lens, respectively. We developed a deconvolution process to reduce scattering-induced crosstalk in multifocal detection scheme, thus enabling whole brain imaging of Drosophila with millisecond and micrometer spatiotemporal resolution. Potential applications toward brain science include studying millisecond dynamics in a neuronal network, and resolving 3D microfluidics in blood vessels.