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dc.contributor.authorCasamayou-Boucau, Yannick
dc.contributor.authorRyder, Alan G.
dc.date.accessioned2018-11-19T12:54:44Z
dc.date.available2018-11-19T12:54:44Z
dc.date.issued2017-08-17
dc.identifier.citationCasamayou-Boucau, Yannick, & Ryder, Alan G. (2017). Extended wavelength anisotropy resolved multidimensional emission spectroscopy (ARMES) measurements: better filters, validation standards, and Rayleigh scatter removal methods. Methods and Applications in Fluorescence, 5(037001). doi: 10.1088/2050-6120/aa7763en_IE
dc.identifier.issn2050-6120
dc.identifier.urihttp://hdl.handle.net/10379/14646
dc.description.abstractAnisotropy resolved multidimensional emission spectroscopy (ARMES) provides valuable insights into multi-fluorophore proteins (Groza et al 2015 Anal. Chim. Acta 886 133-42). Fluorescence anisotropy adds to the multidimensional fluorescence dataset information about the physical size of the fluorophores and/or the rigidity of the surrounding micro-environment. The first ARMES studies used standard thin film polarizers (TFP) that had negligible transmission between 250 and 290 nm, preventing accurate measurement of intrinsic protein fluorescence from tyrosine and tryptophan. Replacing TFP with pairs of broad band wire grid polarizers enabled standard fluorescence spectrometers to accurately measure anisotropies between 250 and 300 nm, which was validated with solutions of perylene in the UV and Erythrosin B and Phloxine B in the visible. In all cases, anisotropies were accurate to better than +/- 1% when compared to literature measurements made with Glan Thompson or TFP polarizers. Better dual wire grid polarizer UV transmittance and the use of excitation-emission matrix measurements for ARMES required complete Rayleigh scatter elimination. This was achieved by chemometric modelling rather than classical interpolation, which enabled the acquisition of pure anisotropy patterns over wider spectral ranges. In combination, these three improvements permit the accurate implementation of ARMES for studying intrinsic protein fluorescence.en_IE
dc.description.sponsorshipThe funders, Irish Research Council (IRC), had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors also acknowledge financial support from an IRC Postgraduate Scholarship (Grant #GOIPG/2015/3826). Agilent Technologies (Australia) are also thanked for a fluorimeter loan.en_IE
dc.formatapplication/pdfen_IE
dc.language.isoenen_IE
dc.publisherIOP Publishingen_IE
dc.relation.ispartofMethods And Applications In Fluorescenceen
dc.subjectfluorescenceen_IE
dc.subjectanisotropyen_IE
dc.subjectmultidimensionalen_IE
dc.subjectstandardsen_IE
dc.subjectpolarizersen_IE
dc.subjectRayleigh scatteren_IE
dc.subjectchemometricsen_IE
dc.subjectMULTIVARIATE CURVE RESOLUTIONen_IE
dc.subjectCELL-CULTURE MEDIAen_IE
dc.subjectFLUORESCENCE-SPECTRAen_IE
dc.subjectEEM SPECTROSCOPYen_IE
dc.subjectSERUM-ALBUMINen_IE
dc.subjectRAMAN SCATTERen_IE
dc.subjectPARAFACen_IE
dc.subjectMOLECULESen_IE
dc.subjectGLYCEROLen_IE
dc.subjectDECOMPOSITIONen_IE
dc.titleExtended wavelength anisotropy resolved multidimensional emission spectroscopy (ARMES) measurements: better filters, validation standards, and Rayleigh scatter removal methodsen_IE
dc.typeArticleen_IE
dc.date.updated2018-11-14T17:25:29Z
dc.identifier.doi10.1088/2050-6120/aa7763
dc.local.publishedsourcehttps://doi.org/10.1088/2050-6120/aa7763en_IE
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funderIrish Research Councilen_IE
dc.internal.rssid13100136
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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