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Antonino Ingargiola edited Abstract.tex
over 9 years ago
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Single-molecule FRET experiments allows to probe the distance between two fluorescent labels (\textit{donor} and \textit{acceptor}) on the 3-10nm range. In the commonly employed confocal geometry, molecules are free to diffuse in a solution. When a molecule crosses the excitation volume it emits a burst of photons that can be detected by single-photon detectors (SPADs). In a nutshell, assessing the relative intensity of the donor and acceptor signal in each bursts it allows to infer their distance. smFRET experiments permit to study binding-unbinding processes or conformational changes of biomolecules and have proven to be an unvaluable tool in studying many cellular processes \cite{Kapanidis_2006}.
The data analysis Analyzing smFRET experiments involves
processing identifing the
photon timestamps single-molecule bursts in
(at least) two spectral channels (donor a continuous stream of photon timestamps, estimating the background and other correction factors, filtering and
acceptor). extracting the corrected FRET efficiencies for each sub-populations in the sample. Although smFRET experiments are routinely performed by many labs around the world, no standard
or reference software exists.
The FRETBursts software
presented described in this paper represents the first
complete opensource
software implementation of all the common state-of-the-art algorithms for smFRET data
analysis implementing most of analysis. We follow the
state-of-the-art algorithms. highest standard in software development to ensure that the source is easy to read, well documented and thoroghly tested. Moreover, in an effort to lower the barriers to computational reproducibility, we embrace a modern workflow based on ipython notebooks that allows to capture the whole process from raw data to figures within a single document.
We aim to build a foundationand heavily documenting the sorce code we strive We strive to provide a software that is easy to use, read and extend. The source code is heavily commented and the algorithms documented and a the reference documentation is automatically build from inline docstrings. Example notebooks and data files are provided to makes easy to get started. The development takes place openly on github, a unit-test suite is automatically run after each commit,