Eitan Lerner edited Burst search.tex  over 9 years ago

Commit id: 6ae1dec4f1eb4d90ce884845ed1c8a7469d134ec

deletions | additions      

       

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')|).