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bioRxiv · 10.1101/2024.03.13.584811

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

Rohr, L.·Ehinger, A.·Burmeister, N. G.·Meixner, A. J.·Kemmerling, B.·Harter, K.·zur Oven-Krockhaus, S.
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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.

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Rohr, L., Ehinger, A., Burmeister, N. G., Meixner, A. J., Kemmerling, B., Harter, K., zur Oven-Krockhaus, S.. 2024-03-15. Simultaneous and Dynamic Super-Resolution Imaging of Two Proteins in Arabidopsis thaliana using dual-color sptPALM. https://doi.org/10.1101/2024.03.13.584811

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