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dc.contributor.authorCasamayou-Boucau, Yannick
dc.contributor.authorRyder, Alan G.
dc.date.accessioned2020-03-12T11:00:36Z
dc.date.available2020-03-12T11:00:36Z
dc.date.issued2017-11-22
dc.identifier.citationCasamayou-Boucau, Yannick, & Ryder, Alan G. (2018). Accurate anisotropy recovery from fluorophore mixtures using Multivariate Curve Resolution (MCR). Analytica Chimica Acta, 1000, 132-143. doi: https://doi.org/10.1016/j.aca.2017.11.031en_IE
dc.identifier.issn1873-4324
dc.identifier.urihttp://hdl.handle.net/10379/15843
dc.description.abstractAnisotropy resolved multidimensional emission spectroscopy (ARMES) provides valuable insights into multi-fluorophore systems like proteins that have complex overlapping emission bands. The method combines multidimensional fluorescence, anisotropy, and chemometrics to facilitate the differentiation of fluorophores with very similar emission properties. Here, we address the critical issue of standardizing the chemometric methods required to accurately extract spectral and anisotropy information from fluorophore mixtures using two standard sample sets: perylene in glycerol, and a mixture of Erythrosin B and Phloxine B with overlapping emission but different anisotropies. We show for the first time how to accurately model component anisotropy using Multivariate Curve Resolution (MCR) from data collected using total synchronous fluorescence scan (TSFS) and Excitation Emission Matrix (EEM) measurement methods.These datasets were selected to avoid the presence of inner filter effects (IFE) or Forster resonance energy transfer (FRET) that would depolarize fluorescence emission or reduce data tri-linearity. This allowed the non-trilinear TSFS data to yield accurate component anisotropy data once modelled using the correct data augmentation strategy, however, the EEM data proved to be more accurate once optimal constraints (non-negativity and correspondence among species) were employed. For perylene (S-2) and Phloxine B which both have very weak anisotropy (0.2) anisotropies, bilinear and trilinear EEM models built using a total tri-linearity constraint, yielded solutions without any rotational ambiguities and very accurate (+/- 4% of real value) anisotropy values. These sample systems thus provide simple and robust test systems for validating the spectral measurement and chemometric data analysis elements of ARMES. (c) 2017 Elsevier B.V. All rights reserved.en_IE
dc.description.sponsorshipYCB was supported by an ‘EMBARK Initiative’ Postgraduate Scholarship (Grant number GOIPG/2015/3826) from the Irish Research Council. We also thank Agilent Technologies (Mulgrave Victoria, Australia) for the loan of a fluorescence spectrometer.en_IE
dc.formatapplication/pdfen_IE
dc.language.isoenen_IE
dc.publisherElsevieren_IE
dc.relation.ispartofAnalytica chimica actaen
dc.subjectFluorescenceen_IE
dc.subjectAnisotropyen_IE
dc.subjectMultidimensionalen_IE
dc.subjectStandardsen_IE
dc.subjectChemometricsen_IE
dc.subject1ST-ORDER RAYLEIGH SCATTERen_IE
dc.subjectEXCITATION-EMISSION DATAen_IE
dc.subjectCELL-CULTURE MEDIAen_IE
dc.subjectFLUORESCENCE SPECTROSCOPYen_IE
dc.subjectEEM SPECTROSCOPYen_IE
dc.subjectSERUM-ALBUMINen_IE
dc.subjectDATA SETSen_IE
dc.subjectPARAFACen_IE
dc.subjectALSen_IE
dc.subjectBILINEARIZATIONen_IE
dc.titleAccurate anisotropy recovery from fluorophore mixtures using Multivariate Curve Resolution (MCR)en_IE
dc.typeArticleen_IE
dc.date.updated2020-03-11T14:56:55Z
dc.identifier.doi10.1016/j.aca.2017.11.031
dc.local.publishedsourcehttps://doi.org/10.1016/j.aca.2017.11.031en_IE
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funderIrish Research Councilen_IE
dc.internal.rssid13714146
dc.local.contactAlan Ryder, School Of Chemistry, Room 213, Arts/Science Building, South Cam, Nui Galway. 2943 Email: alan.ryder@nuigalway.ie
dc.local.copyrightcheckedYes
dc.local.versionACCEPTED
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