 |
Center for Nonlinear Studies |
In order to understand how brains compute at a mechanistic level it will be necessary to record the activity of large populations of neurons simultaneously. Given the immense scale and fast dynamics of the nervous system this requires advances in both hardware and software. We advance the state-of-theart with a novel light-sheet microscope designed to maximize the volume imaging rate of the Objective-Coupled Planar Illumination (OCPI) family of microscopes. OCPI microscopes, like most light-sheet microscopes, are rate-limited by two aspects of their design: the mechanics of scanning and the maximum framerate of modern cameras. We address these bottlenecks by optimizing scan mechanics and introducing Distributed Planar Imaging (DPI), a technique that allows framerate to scale with the number of cameras used. Our design is unique among fast imaging methods in that it requires no sacrifice of image quality or photon efficiency when compared to conventional light-sheet microscopies. We demonstrate fast (10-20Hz) 3D imaging of calcium dynamics in thousands of neurons throughout the larval zebrafish brain at 0.65μm x 0.65μm x 5μm resolution. We also provide m x 0.65μm x 0.65μm x 5μm resolution. We also provide m x 5μm x 0.65μm x 5μm resolution. We also provide m resolution. We also provide an overview of our preprocessing and analysis pipeline developed for these multi-terabyte datasets.
Host: Anatoly Zlotnik