A long term time lapse microscopy technique for Arabidopsis roots

TL;DR

This paper introduces a new light sheet microscopy method for long-term imaging of Arabidopsis roots, improving accessibility and reducing the need for special software.

Contribution

A novel protocol for sample mounting and automated root tip tracking in light sheet microscopy for plant studies.

Findings

A new sample mounting protocol was developed for Arabidopsis roots in light sheet microscopy.

An automated root tip tracking system was created without requiring proprietary software.

The method enables long-term time-lapse imaging of regenerating Arabidopsis roots after laser ablation.

Abstract

Time lapse microscopy is a transformative technique for plant cell and developmental biology. Light sheet microscopy, which manipulates the amount of light a sample is exposed to in order to minimize phototoxicity and maximize signal intensity, is an increasingly popular tool for time lapse imaging. However, many light sheet imaging systems are not designed with the unique properties of plant samples in mind. Recent advances have decreased the cost and increased the technical accessibility of light sheet microscopy, but plant samples still require special preparation to be compatible with these new systems. Here, we apply a novel light sheet microscopy system to regenerating Arabidopsis roots damaged via laser ablation. To adapt this system for Arabidopsis roots we establish a new protocol for sample mounting, as well as an automated root tip tracking system that requires no additional proprietary software. The methods presented here can be used to increase researcher access to long-term time-lapse imaging in Arabidopsis biology.

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