Revealing Plasma Membrane Protein Dynamics in Living Plant Cells with Single-Molecule Tracking

The behavior of proteins and other biomolecules in cellular environments is governed by complex molecular processes. Understanding their temporal dynamics and interactions with other biomolecules or cellular structures is essential for elucidating physiological functions. Single-particle tracking photoactivated localization microscopy (sptPALM) has emerged as a powerful single-molecule technique for investigating these processes with exceptional spatial and temporal resolution. In this Expert View, we introduce sptPALM with a special focus on its application in the plasma membrane of plant cells. Key aspects and advances of the technique, including instrumentation and data analysis, are discussed to equip researchers with the foundational knowledge required to establish and execute sptPALM experiments. Recent studies are highlighted to demonstrate the potential of sptPALM to advance our understanding of molecular dynamics in plant cells.