DOI: 10.1101/501890Dec 19, 2018Paper

Fast Objective Coupled Planar Illumination Microscopy

BioRxiv : the Preprint Server for Biology
Cody J Greer, Timothy E Holy

Abstract

Among optical imaging techniques light sheet fluorescence microscopy stands out as 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 current-generation light sheet microscopes are limited by volume scanning rate and/or camera frame rate. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique to avoid compromising image quality or photon efficiency. We demonstrate imaging rates up to 40 volumes per second, and tissue volumes up to 700 μm thick, and introduce Multi-Camera Image Sharing (MCIS), a technique to scale imaging rate by parallelizing acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact that can be removed by filtering when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.

Related Concepts

Brain
Diagnostic Imaging
Microscopy
Microscopy, Fluorescence
Zebrafish
Surgical Drapes
Heart Beat
Scanning
Illumination Technique
Monitoring - Action

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