L. Schrangl, J. Göhring, G. Schütz:

"Kinetic analysis of single molecule FRET transitions without trajectories";

Journal of Chemical Physics,148(2018), 1233281 - 1233289.

Single molecule F¨orster resonance energy transfer (smFRET) is a popular tool to study biological

systems that undergo topological transitions on the nanometer scale. smFRET experiments typically

require recording of long smFRET trajectories and subsequent statistical analysis to extract parameters

such as the states´ lifetimes. Alternatively, analysis of probability distributions exploits the shapes

of smFRET distributions at well chosen exposure times and hence works without the acquisition

of time traces. Here, we describe a variant that utilizes statistical tests to compare experimental

datasets with Monte Carlo simulations. For a given model, parameters are varied to cover the full

realistic parameter space. As output, the method yields p-values which quantify the likelihood for each

parameter setting to be consistent with the experimental data. The method provides suitable results

even if the actual lifetimes differ by an order of magnitude.We also demonstrated the robustness of the

method to inaccurately determine input parameters. As proof of concept, the new method was applied

to the determination of transition rate constants for Holliday junctions.

Created from the Publication Database of the Vienna University of Technology.