Quantitation of Cellular Dynamics in Growing Arabidopsis Roots with Light Sheet Microscopy

Abstract

To understand dynamic developmental processes, living tissues must be imaged frequently and for extended periods of time. Root development is extensively studied at cellular resolution to understand basic mechanisms underlying pattern formation and maintenance in plants. Unfortunately, ensuring continuous specimen access, while preserving physiological conditions and preventing photo-damage, poses major barriers to measurements of cellular dynamics in indeterminately growing organs such as plant roots. We present a system that integrates optical sectioning through light sheet fluorescence microscopy with hydroponic culture that enables us to image at cellular resolution a vertically growing Arabidopsis root every few minutes and for several consecutive days. We describe novel automated routines to track the root tip as it grows, track cellular nuclei and identify cell divisions. We demonstrate the system's capabilities by collecting data on divisions and nuclear dynamics.