TY - JOUR T1 - FluoSim: simulator of single molecule dynamics for fluorescence live-cell and super-resolution imaging of membrane proteins JF - bioRxiv DO - 10.1101/2020.02.06.937045 SP - 2020.02.06.937045 AU - Matthieu Lagardère AU - Ingrid Chamma AU - Emmanuel Bouilhol AU - Macha Nikolski AU - Olivier Thoumine Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/02/06/2020.02.06.937045.abstract N2 - Fluorescence live-cell and super-resolution microscopy methods have considerably advanced our understanding of the dynamics and mesoscale organization of macro-molecular complexes that drive cellular functions. However, different imaging techniques can provide quite disparate information about protein motion and organization, owing to their respective experimental ranges and limitations. To address these limitations, we present here a unified computer program that allows one to model and predict membrane protein dynamics at the ensemble and single molecule level, so as to reconcile imaging paradigms and quantitatively characterize protein behavior in complex cellular environments. FluoSim is an interactive real-time simulator of protein dynamics for live-cell imaging methods including SPT, FRAP, PAF, and FCS, and super-resolution imaging techniques such as PALM, dSTORM, and uPAINT. The software, thoroughly validated against experimental data on the canonical neurexin-neuroligin adhesion complex, integrates diffusion coefficients, binding rates, and fluorophore photo-physics to calculate in real time the distribution of thousands of independent molecules in 2D cellular geometries, providing simulated data of protein dynamics and localization directly comparable to actual experiments. ER -