Simultaneous and Dynamic Super-Resolution Imaging of Two Proteins in Arabidopsis thaliana using dual-color sptPALM

Abstract

Super-resolution microscopy techniques have revolutionized cell biology by providing insights into single-molecule dynamics and nanoscale organization within living cells. However, the application of dynamic live-cell methods in plants remains limited by the availability of suitable fluorophores for simultaneous visualization of multiple proteins. To address this challenge, we implemented a dual-color single-particle tracking photoactivated localization microscopy (sptPALM) approach based on codon-optimized photoactivatable fluorescent proteins PA-GFP and PATagRFP. Recently, we demonstrated their individual performance in single-color experiments in Nicotiana benthamiana and Arabidopsis thaliana cells. Here, we establish their combined use for dual-color sptPALM, enabling the simultaneous tracking of two distinct protein species within the same plant cell. This approach provides a framework to investigate the coordinated dynamics, interactions, and spatial organization of multiple proteins in living plant cells.

Plant phenotyping relevance

植物細胞内の2種類のタンパク質を同時追跡するデュアルカラーsptPALM法を開発・確立した研究であり、生細胞の動態・空間配置という植物状態の取得法が中心です。

Code and data availability

植物フェノタイピング解析を再現する公開資産であることを、入力本文と直接リンクから確認できなかったため保留しました。

No evidence-backed public reproduction asset is currently recorded.