Mesoscale volumetric light field (MesoLF) imaging of neuroactivity across cortical areas at 18 Hz.

Various implementations of mesoscopes provide optical access for calcium imaging across multi-millimeter fields-of-view (FOV) in the mammalian brain. However, capturing the activity of the neuronal population within such FOVs near-simultaneously and in a volumetric fashion has remained challenging since approaches for imaging scattering brain tissues typically are based on sequential acquisition. Here, we present a modular, mesoscale light field (MesoLF) imaging hardware and software solution that allows recording from thousands of neurons within volumes of ⍰ 4000 × 200 µm, located at up to 400 µm depth in the mouse cortex, at 18 volumes per second. Our optical design and computational approach enable up to hour-long recording of ∼10,000 neurons across multiple cortical areas in mice using workstation-grade computing resources.