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dc.contributor.authorMelnikau, Dzmitry
dc.contributor.authorElcoroaristizabal, Saioa
dc.contributor.authorRyder, Alan G.
dc.date.accessioned2018-11-19T14:43:14Z
dc.date.issued2018-09-03
dc.identifier.citationMelnikau, Dzmitry, Elcoroaristizabal, Saioa, & Ryder, Alan G. (2018). An excitation emission fluorescence lifetime spectrometer using a frequency doubled supercontinuum laser source. Methods and Applications in Fluorescence, 6 (045007). doi: 10.1088/2050-6120/aad9aeen_IE
dc.identifier.issn2050-6120
dc.identifier.urihttp://hdl.handle.net/10379/14649
dc.description.abstractThe accurate fluorescence analysis of complex, multi-fluorophore containing proteins requires the use of multi-dimensional measurement techniques. For the measurement of intrinsic fluorescence from tyrosine (Tyr) and tryptophan (Trp) one needs tuneable UV excitation and for steady-state measurements like Excitation Emission Matrix (EEM) simple pulsed Xe lamps are commonly used. Unfortunately, simultaneous multi-dimensional wavelength and time resolved measurement of intrinsic protein fluorescence in the 260 to 400 nm spectral range are challenging and typically required the use of very complex tuneable laser systems or multiple single excitation wavelength sources. Here we have assembled and validated a novel Excitation Emission Fluorescence Lifetime Spectrometer (EEFLS) using a pulsed, frequency doubled, Super-Continuum Laser (SCL) source coupled with a 16 channel multi-anode Time Correlated Single Photon Counting (TCSPC) measurement system. This EEFLS enabled the collection of near complete lifetime and intensity maps over the most important intrinsic protein fluorescence spectral range (lambda(ex) = 260-350/lambda(em) = 300-500 nm). The 4-dimensional (lambda(ex)/lambda(em)/I-(t)/tau) Excitation Emission Fluorescence Lifetime Matrix (EEFLM) data produced can be used to better characterize the complex intrinsic emission from proteins. The system was capable of measuring fluorescence emission data with high spectral (1-2 nm) resolution and had an Instrument Response Function (IRF) of similar to 650 ps for accurate measurement of nanosecond lifetimes. UV power output was stable after a warm up period, with variations ofen_IE
dc.description.sponsorshipThis publication has emanated from research supported in part by a research grant from Science Foundation Ireland (SFI) and is co-funded under the European Regional Development Fund under Grand number (14/IA/2282, Advanced Analytics for Biological Therapeutic Manufacture, to AGR). The authors declare that there are no conflicting interests.en_IE
dc.formatapplication/pdfen_IE
dc.language.isoenen_IE
dc.publisherIOP Publishingen_IE
dc.relation.ispartofMethods And Applications In Fluorescenceen
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Ireland
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/ie/
dc.subjectfluorescenceen_IE
dc.subjectsupercontinuumen_IE
dc.subjectlifetimeen_IE
dc.subjectexcitation emission matrixen_IE
dc.subjectfrequency doubleden_IE
dc.subjectCELL-CULTURE MEDIAen_IE
dc.subjectTIME-RESOLVED FLUORESCENCEen_IE
dc.subjectHETEROGENEOUS FLUORESCENCEen_IE
dc.subjectQUANTITATIVE-ANALYSISen_IE
dc.subjectTRYPTOPHANen_IE
dc.subjectPROTEINSen_IE
dc.subjectRESOLUTIONen_IE
dc.subjectSPECTROSCOPYen_IE
dc.subjectPHASEen_IE
dc.subjectDECAYen_IE
dc.titleAn excitation emission fluorescence lifetime spectrometer using a frequency doubled supercontinuum laser sourceen_IE
dc.typeArticleen_IE
dc.date.updated2018-11-14T17:57:20Z
dc.identifier.doi10.1088/2050-6120/aad9ae
dc.local.publishedsourcehttps://doi.org/10.1088/2050-6120/aad9aeen_IE
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funderScience Foundation Irelanden_IE
dc.description.embargo2019-09-03
dc.internal.rssid14946835
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
dcterms.projectinfo:eu-repo/grantAgreement/SFI/SFI Investigator Programme/14/IA/2282/IE/Advanced Analytics for Biological Therapeutic Manufacture (AA-BTM)./en_IE
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Attribution-NonCommercial-NoDerivs 3.0 Ireland
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 3.0 Ireland