A long term time lapse microscopy technique for Arabidopsis roots.

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.

Plant phenotyping relevance

Arabidopsis根の長期タイムラプス撮像系を植物試料向けに適応し、試料マウント法と自動根端追跡を開発しており、表現型取得・抽出法が中心である。

Code and data availability

The paper describes a light-sheet time-lapse method for Arabidopsis roots with custom FIJI/VisiView tracking macros and raw imaging data, but no public deposit or authors' URL for code or data is given. Raw data are available only on request, and the supplementary material contains only screenshots of macro execution,

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