A flexible approach to assess fluorescence decay functions in complex energy transfer systems

摘要: Time-correlated Forster resonance energy transfer (FRET) probes molecular distances with greater accuracy than intensity-based calculation of FRET efficiency and provides a powerful tool to study biomolecular structure dynamics. Moreover, time-correlated photon count measurements bear additional information on the variety donor surroundings allowing more detailed differentiation between distinct structural geometries which are typically inaccessible general fitting solutions. Here we develop new approach based Monte Carlo simulations events estimate single counts (TCSPC) histograms in complex systems. This simulation solution assesses full statistics distance distributions fluorescently labeled biomolecules. The consistent theoretical predictions dye behavior systems defined randomly distributed We validate results using highly heterogeneous aggregation system explore conditions use this is distinguishing wide experimental setups, thus providing versatile accurately distinguish different assemblies