Two-photon Bessel beam tomography for fast volume imaging

Abstract

Light microscopy on dynamic samples, for example neural activity in the brain, often requires imaging volumes that extend over several 100 µm in axial direction at a rate of at least several tens of Hertz. Here, we develop a tomography approach for scanning fluorescence microscopy which allows recording a volume image in a single frame scan. Volumes are imaged by simultaneously recording four independent projections at different angles using temporally multiplexed, tilted Bessel beams. From the resulting projections, three-dimensional images are reconstructed using inverse Radon transforms combined with convolutional neural networks (U-net).