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Antonino Ingargiola edited Burst_Weights_Theory.tex
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the accompanying Jupyter notebook (\href{}{link}).
\subsubsection{Comparison FRET histograms}
The
second simulation, is much more realistic and captures effect of weighting the
complexity FRET histogram is here illustrated with a simulation of
a mixture of two static FRET populations and then with experimental
smFRET data.
Starting We performed a realistic simulation of a static mixture of two FRET populations
starting from 3-D Brownian motion diffusion of $N$ particles excited by a
numerically computed PSF, we simulated timestamps
of a smFRET experiments with two static FRET populations. numerically-computed (non-Gaussian PSF). Input parameters includes
the diffusion coefficient, the particle brightness, the two FRET efficiencies,
as well as detectors DCR. The simulation is performed with the open source software
PyBroMo~\cite{Ingargiola_2016} which creates smFRET data files
(i.e. timestamps and detectors
arrays) in Photon-HDF5 format~\cite{Ingargiola2016}.
After a The simulated data file is processed with FRETBursts performing burst search,
and only a minimal burst size selection of with threshold of 10
photons,
we obtained the photons.
The resulting weighted and unweighted FRET histograms
are reported in figure~\ref{fig:YYY}.
We notice as the use of the weights results in better definition of FRET peaks.
As a final comparison, we report the weighted and unweighted FRET histogram of
an experimental FRET population from measurement of a di-labeled dsDNA sample.
Figure X show a comparison of a FRET histogram obtained from the same burst
with and without weights. The burst selection is obtained applying a burst size
of threshold of 10 counts (after background correction), in order to filter
the extreme low-end of the burst size distribution.
The use of size-weighted FRET histograms is a simple way to obtain a representation of FRET
distribution that maintains high power of resolving
FRET peaks while including the full burst
population and thus reducing statistical noise.
As a final remark, note that when increasing the size-threshold for burst selection