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Journal Article
Fast objective coupled planar illumination microscopy
2019
Overview
Among optical imaging techniques light sheet fluorescence microscopy is one of the most attractive for capturing high-speed biological dynamics unfolding in three dimensions. The technique is potentially millions of times faster than point-scanning techniques such as two-photon microscopy. However light sheet microscopes are limited by volume scanning rate and/or camera speed. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique that avoids compromising image quality or photon efficiency. Our fast scan system supports 40 Hz imaging of 700 μm-thick volumes if camera speed is sufficient. We also address the camera speed limitation by introducing Distributed Planar Imaging (DPI), a scaleable technique that parallelizes image acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact, removable when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.
Light sheet microscopy holds potential for imaging dynamics in 3D biological specimens, but is limited by scan speed and camera acquisition rate. Here the authors address both issues by developing speed-optimized Objective Coupled Planar Illumination and parallelizing image acquisition across cameras to achieve 40 Hz imaging over thick samples.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 631/378
/ 64/116
/ 96/35
/ Animals
/ Cameras
/ Diagnostic Imaging - instrumentation
/ Diagnostic Imaging - methods
/ Humanities and Social Sciences
/ Image Processing, Computer-Assisted - instrumentation
/ Image Processing, Computer-Assisted - methods
/ Larva
/ Light
/ Luminescent Measurements - instrumentation
/ Luminescent Measurements - methods
/ Microscopy - instrumentation
/ Microscopy, Confocal - instrumentation
/ Microscopy, Confocal - methods
/ Microscopy, Fluorescence - instrumentation
/ Microscopy, Fluorescence - methods
/ Microscopy, Fluorescence, Multiphoton - instrumentation
/ Microscopy, Fluorescence, Multiphoton - methods
/ Photons
/ Scanning
/ Science
