This notebook performs realtime kinetics data analysis for the paper:
Multi-spot single-molecule FRET: high-throughput analysis of freely diffusing molecules
Ingargiola et al. PLOS ONE (2017)
doi:10.1371/journal.pone.0175766
For the static dsDNA analysis please refer to:
The realtime kinetics dataset can be downloaded from:
Before running the analysis please put these data files in the realtime kinetics/data
subfolder.
The following notebook analyzes the raw multispot realtime kinetics smFRET data for the different measurement replicates. Burst search results and FRET efficiencies fits are computed on a moving time window:
After this preprocessing the data is summarized and plotted in:
Similarly, single-spot μs-ALEX smFRET on the same sample is analyzed in:
And summarized in:
Simulations have been performed to study the fitting accuracy of each parameter when using the simple-exponential and integrated-exponential models for a range of conditions. We test noise levels typical for both multispot and single-spot measurements. For each noise level we performed the model comparison allowing $t_0$ (the origin of the transient) to vary or not.
All these simulations are performed in:
Finally, the notebook:
studies how the variance of a Gaussian random process scales with the size of a moving integration window and the effect of overlapping or non-overlapping windows.