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\section{Architecture and Concepts}
\label{sec:concepts}
In this section, we introduce some general
burst analysis concepts
and notations used in FRETBursts.
\subsection{Photon Streams}
\label{sec:ph_streams}
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In single-spot measurements, all timestamps are stored in a single array. In multispot
measurements~\cite{Ingargiola_2013}, there are as many timestamps arrays
as excitation spots.
Each array contains timestamps from both donor (D) and acceptor (A) channels.
When alternating excitation lasers are used (ALEX measurements)~\cite{Lee_2005},
a further distinction between photons emitted during the D or A excitation periods can be made.
In FRETBursts, the corresponding sets of photons are called ``photon streams'' and are
specified with a \verb|Ph_sel| object
(\href{http://fretbursts.readthedocs.org/en/latest/ph_sel.html}{link}).
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\label{sec:bg_intro}
An estimation of the background rates is needed to both select a proper threshold for
burst search, and
to correct the raw burst counts by subtraction of background counts.
The recorded stream of timestamps is the result of two processes: one characterized
by a high count rate, due to fluorescence photons of single molecules crossing the
excitation volume, and another characterized by a lower count rate, due to
"background
counts" ``background
counts'' originating from detector dark counts, afterpulsing, out-of-focus molecules
and sample scattering and/or impurities~\cite{Edman_1996,Gopich_2008}.
The signature of these two types of processes can be
observed in the inter-photon delays distribution (i.e. the waiting times
between two subsequent timestamps) as illustrated in figure~\ref{fig:bg_dist_all}(a).
The
“tail” ``tail'' of the distribution (a straight line in semi-log scale) corresponds
to exponentially-distributed time-delays, indicating that those counts are generated by a
Poisson process. At short
timescales, the distribution departs from the exponential due to the contribution
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FRETBursts allows performing the burst search on arbitrary selections of photons.
(see section~\ref{sec:ph_streams} for more information on photon stream definitions).
Additionally, Nir~\textit{et al.}~\cite{Nir_2006} proposed DCBS
('dual-channel (``dual-channel burst
search'), search''), which can help mitigating artifacts due to photophysics effects such as blinking.
During DCBS, a search is performed in parallel on two photon streams
and bursts are defined as periods during which both photon streams exhibit a rate higher than
the threshold, implementing the equivalent of an AND logic operation.
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are (1) all photons during donor excitation (\verb|Ph_sel(Dex='DAem')|) and
(2) acceptor channel photons during acceptor excitation (\verb|Ph_sel(Aex='Aem')|).
In FRETBursts, the user can choose arbitrary photon streams as input, an in general
this kind of search is called a
'AND-gate ``AND-gate burst
search'. search''.
After burst search, it is necessary to select
bursts, for instance by specifying a minimum number of photons (or burst size). In the most
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the burst size selection threshold is weighting bursts proportionally to their size
so that the bursts with largest sizes will have the largest weights.
Using size as weights (instead of any other monotonically increasing function
of size) can be justified noticing that the variance of bursts
PR ($E_i$) proximity ratio (PR) is
inversely proportional to the burst size (see SI~\ref{sec:burstweights_theory} for details).
In general, a weighting scheme is used for building efficient estimators for a population
parameter (e.g.
the population FRET efficiency $E_p$).
But, it can also be used to build weighted histograms or Kernel Density
Estimation (KDE) plots which emphasize FRET subpopulations peaks
without excluding small size bursts.
Traditionally, for optimal results when not using weights, the
diff --git a/figures/ph_delays_distrib_all/caption.tex b/figures/ph_delays_distrib_all/caption.tex
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(\textit{Panel a}) An example of inter-photon delays distribution (\textit{red dots}) and an exponential fit
of the tail of the distribution (\textit{black line}).
(\textit{Panel b}) Inter-photon delays distribution and exponential fit for different photon streams as obtained with \texttt{dplot(d, hist\_bg)}. The \textit{dots} represent the experimental histogram for the different photon streams. The \textit{solid lines} represent the corresponding exponential fit of the tail of the distributions. The legend shows abbreviations of the photon streams
and the fitted
rate background
rate. rates.