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dc.contributor.authorLaine, Romain F
dc.contributor.authorPoudel, Chetan
dc.contributor.authorKaminski, Clemens F
dc.date.accessioned2022-06-29T19:44:49Z
dc.date.available2022-06-29T19:44:49Z
dc.date.issued2022-09
dc.date.submitted2021-09-02
dc.identifier.issn0022-2720
dc.identifier.otherjmi13128
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/338489
dc.description.abstractFluorescence lifetime imaging (FLIM) allows the quantification of sub-cellular processes in situ, in living cells. A number of approaches have been developed to extract the lifetime from time-domain FLIM data, but they are often limited in terms of speed, photon efficiency, precision or the dynamic range of lifetimes they can measure. Here, we focus on one of the best performing methods in the field, the centre-of-mass method (CMM), that conveys advantages in terms of speed and photon efficiency over others. In this paper, however, we identify a loss of photon efficiency of CMM for short lifetimes when background noise is present. We subsequently present a new development and generalization of CMM that provides for the rapid and accurate extraction of fluorescence lifetime over a large lifetime dynamic range. We provide software tools to simulate, validate and analyse FLIM data sets and compare the performance of our approach against the standard CMM and the commonly employed least-square minimization (LSM) methods. Our method features a better photon efficiency than standard CMM and LSM and is robust in the presence of background noise. The algorithm is applicable to any time-domain FLIM data set.
dc.languageen
dc.publisherWiley
dc.subjectORIGINAL ARTICLE
dc.subjectORIGINAL ARTICLES
dc.subjectcenter of mass
dc.subjectFiji plugin
dc.subjectFLIM
dc.subjectquantitative analysis
dc.titleA method for the fast and photon-efficient analysis of time-domain fluorescence lifetime image data over large dynamic ranges.
dc.typeArticle
dc.date.updated2022-06-29T19:44:48Z
prism.publicationNameJ Microsc
dc.identifier.doi10.17863/CAM.85902
dcterms.dateAccepted2022-05-31
rioxxterms.versionofrecord10.1111/jmi.13128
rioxxterms.versionAO
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidLaine, Romain F [0000-0002-2151-4487]
dc.contributor.orcidPoudel, Chetan [0000-0002-8512-9238]
dc.contributor.orcidKaminski, Clemens F [0000-0002-5194-0962]
dc.identifier.eissn1365-2818
pubs.funder-project-idEngineering and Physical Sciences Research Council (EP/H018301/1)
pubs.funder-project-idWellcome Trust (089703/Z/09/Z)
pubs.funder-project-idMedical Research Council (MR/K015850/1)
pubs.funder-project-idMedical Research Council (MR/K02292X/1)
pubs.funder-project-idEngineering and Physical Sciences Research Council (EP/L015889/1)
pubs.funder-project-idEuropean Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (722380)
cam.issuedOnline2022-06-23


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