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bioRxiv · 10.1101/2023.08.10.552827

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

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

Super-resolution microscopy (SRM) approaches revolutionize cell biology by providing insights into the nanoscale organization and dynamics of macromolecular assemblies and single molecules in living cells. A major hurdle limiting SRM democratization is post-acquisition data analysis which is often complex and time-consuming. Here, we present OneFlowTraX, a user-friendly and open-source software dedicated to the analysis of single-molecule localization microscopy (SMLM) approaches such as single-particle tracking photoactivated localization microscopy (sptPALM). Through an intuitive graphical user interface, OneFlowTraX provides an automated all-in-one solution for single-molecule localization, tracking, as well as mobility and clustering analyses. OneFlowTraX allows the extraction of diffusion and clustering parameters of millions of molecules in a few minutes. Finally, OneFlowTraX greatly simplifies data management following the FAIR (Findable, Accessible, Interoperable, Reusable) principles. We provide a detailed step-by-step manual and guidelines to assess the quality of single-molecule analyses. Applying different fluorophores including mEos3.2, PA-GFP, and PATagRFP, we exemplarily used OneFlowTraX to analyze the dynamics of plant plasma membrane-localized proteins including an aquaporin, the brassinosteroid receptor Brassinosteroid Insensitive 1 (BRI1) and the Receptor-Like Protein 44 (RLP44).

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Rohr, L., Ehinger, A., Rausch, L., Burmeister, N. G., Meixner, A. J., Harter, K., Kemmerling, B., zur Oven-Krockhaus, S.. 2023-08-11. GreenMemTraX: A User-Friendly Software for Super-Resolution Analysis of Membrane Protein Dynamics and Nanoscale Organization in Plants. https://doi.org/10.1101/2023.08.10.552827

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