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Eitan Lerner edited Burst search.tex
over 9 years ago
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After background estimation, the burst search is the next fundamental step of the analysis. The core "sliding window" algorithm, proposed by Eggeling~\textit{et al.} in 1998~\cite{Eggeling_1998}, involves searching for bursts of photons
in which $m$ consecutive photons are contained within a minimal time period $\Delta t$. In other words, bursts are portions of the photon stream where the local rate (computed using $m$ photons) is above a minimal rate chosen as a threshold. Eggeling did not provide any criteria on how to choose the rate threshold and the number of photons $m$ and as therefore it has become a common practice to manually tweak those parameters for each specific measurement.
A more general approach consist in taking into account the background rate of the specific measurements and in choosing a rate threshold that is $F$ times larger than the background rate. This approach assures that the resulting bursts all have a single-to-background ratio (SBR) larger than $(F-1)$~\cite{Michalet_2012}. A consistent
criterium criterion to choose the threshold is very important when comparing different measurements with different background rates, when the background significantly changes during the measurements or in multi-spot measurements where each spot has a different background
rates. rate.
A second important aspect of burst search is which photon stream is processed.
Usually, Sometimes, for instance when identifying FRET populations,
we want one would like to apply the burst search to all the photons. Other times, when focusing on donor-only or acceptor only
population, populations, it is better to use only the donor or acceptor signal. In general
we want one would like to be able to apply the burst search to an arbitrary selection of photons. In FRETBursts this can be achieved passing the appropriate \verb|Ph_sel| object to the burst search method (see section~\ref{sec:ph_streams} for more info on photon stream definitions).
Finally, Nir~\textit{et al.}~\cite{Nir_2006} proposed a dual-channel burst search (DCBS) that allows
one to mitigate
(to some extent) artifacts due to photo-physical effects such as blinking. In this case a search is
performer performed independently on two photon streams and bursts are marked only when both photon streams exhibit a rate higher than the threshold,
implementing a kind of
an AND-gate logic.
Usually, the term DCBS is refers to a burst search where the two photon streams are all the photons
during donor excitation (\verb|Ph_sel(Dex='DAem')|) and acceptor channel photons during acceptor
excitation (\verb|Ph_sel(Aex='Aem')|).