Stochastic approach to the molecular counting problem in superresolution microscopy.

dc.contributor.authorRollins, Geoffrey C.
dc.contributor.authorYen Shin, Jae
dc.contributor.authorBustamante, Carlos
dc.contributor.authorPressé, Steve
dc.contributor.departmentDepartment of Physics, School of Scienceen_US
dc.date.accessioned2016-02-29T13:58:59Z
dc.date.available2016-02-29T13:58:59Z
dc.date.issued2015-01-13
dc.description.abstractSuperresolution imaging methods--now widely used to characterize biological structures below the diffraction limit--are poised to reveal in quantitative detail the stoichiometry of protein complexes in living cells. In practice, the photophysical properties of the fluorophores used as tags in superresolution methods have posed a severe theoretical challenge toward achieving this goal. Here we develop a stochastic approach to enumerate fluorophores in a diffraction-limited area measured by superresolution microscopy. The method is a generalization of aggregated Markov methods developed in the ion channel literature for studying gating dynamics. We show that the method accurately and precisely enumerates fluorophores in simulated data while simultaneously determining the kinetic rates that govern the stochastic photophysics of the fluorophores to improve the prediction's accuracy. This stochastic method overcomes several critical limitations of temporal thresholding methods.en_US
dc.identifier.citationRollins, G. C., Shin, J. Y., Bustamante, C., & Pressé, S. (2015). Stochastic approach to the molecular counting problem in superresolution microscopy. Proceedings of the National Academy of Sciences of the United States of America, 112(2), E110–E118. http://doi.org/10.1073/pnas.1408071112en_US
dc.identifier.urihttps://hdl.handle.net/1805/8552
dc.language.isoen_USen_US
dc.publisherPNASen_US
dc.relation.isversionof10.1073/pnas.1408071112en_US
dc.relation.journalProceedings of the National Academy of Sciences of the United States of Americaen_US
dc.rightsPublisher Policyen_US
dc.sourcePMCen_US
dc.subjectCounting problemen_US
dc.subjectFluorescenceen_US
dc.subjectProtein complexesen_US
dc.subjectSingle moleculeen_US
dc.subjectSuperresolutionen_US
dc.titleStochastic approach to the molecular counting problem in superresolution microscopy.en_US
dc.typeArticleen_US
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