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

Super-resolution microscopy techniques have revolutionized cell biology by providing insights into the dynamics of single molecules and nanoscale organization within living cells. However, the application of dynamic live-cell methods in plants has been limited by the lack of suitable fluorophores for simultaneous visualization of multiple proteins. To address this challenge, we implemented a two-color sptPALM approach using codon-optimized photoactivatable fluorescent proteins PA-GFP and PATagRFP. Recently, we showed their individual usability in single-color experiments in Nicotiana benthamiana and Arabidopsis thaliana cells. Here, we now demonstrate the suitability of these fluorophores and their combined use for dual-color sptPALM for the simultaneous observation of two different protein fusions in the same plant cell.