GreenMemTraX: A User-Friendly Software for Super-Resolution Analysis of Membrane Protein Dynamics and Nanoscale Organization in Plants

Super-resolution microscopy provides deep insights into the dynamics of single membrane proteins and the nanoscale organization of plasma membrane proteins in living cells. However, in addition to plant-specific challenges in imaging applications, the elaborate analysis of single-molecule data complicates its widespread use in plant cell biological research. With GreenMemTraX, we provide a user-friendly, all-in-one, open-source software package that guides users through the steps of localization, tracking, mobility, and cluster analyses of plant plasma membrane proteins based on single particle tracking Photoactivated Localization Microscopy (sptPALM) data. GreenMemTraX dramatically simplifies data management following the FAIR (Findable, Accessible, Interoperable, Reusable) principles and increases the ease of use and reproducibility of the evaluated results. In addition, batch processing with detailed parameter output is available for all analysis paths, maintaining the consistency of raw data with the calculated results. The applicability of GreenMemTraX is demonstrated by evaluating sptPALM data for two representative plant plasma membrane proteins, namely the brassinosteroid receptor Brassinosteroid Insensitive 1 (BRI1) and the Receptor-Like Protein 44 (RLP44). Both were tested with the photoconvertible or photoactivatable fluorophores mEos3.2, PA-GFP, and PATagRFP and expressed in the Nicotiana benthamiana transient expression system and in stably transformed Arabidopsis thaliana plants. To facilitate plant applications, GreenMemTraX provides a number of preset parameter settings to address plant-specific obstacles. ### Competing Interest Statement The authors have declared no competing interest.