FRET Imaging of Hemoglobin Concentration in Plasmodium falciparum-Infected Red Cells 英文参考文献.docVIP

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FRET Imaging of Hemoglobin Concentration in Plasmodium falciparum-Infected Red Cells 英文参考文献.doc

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FRET Imaging of Hemoglobin Concentration in Plasmodium falciparum-Infected Red Cells 英文参考文献

FRETImagingofHemoglobinConcentrationin Plasmodiumfalciparum-InfectedRedCells AlessandroEsposito1,2*,TeresaTiffert2,JakobM.A.Mauritz1,2,SimonSchlachter1,LawrenceH. Bannister3,ClemensF.Kaminski1,4,VirgilioL.Lew2 1Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, United Kingdom, 2Department of Physiology, Development and Neuroscience,UniversityofCambridge,Cambridge,UnitedKingdom,3CentreforUltrastructuralImaging,KingsCollegeLondon,London,UnitedKingdom,4Schoolof AdvancedOpticalTechnologies,Max-Planck-ResearchGroup,DivisionIII,UniversityofErlangen-Nuremberg,Erlangen,Germany Abstract Background: During its intraerythrocytic asexual reproduction cycle Plasmodium falciparum consumes up to 80% of the hostcellhemoglobin,inlargeexcessoveritsmetabolicneeds.Amodelofthehomeostasisoffalciparum-infectedredblood cellssuggestedanexplanationbasedontheneedtoreducethecolloid-osmoticpressurewithinthehostcelltopreventits premature lysis. Critical for this hypothesis was that the hemoglobin concentration within the host cell be progressively reducedfromthetrophozoitestageonwards. Methodology/Principal Findings: The experiments reported here were designed to test this hypothesis by direct measurementsofthehemoglobinconcentrationinlive,infectedredcells.Wedevelopedanovel,non-invasivemethodto quantifythehemoglobinconcentrationinsinglecells,basedonFo¨rsterresonanceenergytransferbetweenhemoglobin moleculesandthefluorophorecalcein.Fluorescencelifetimeimagingallowedthequantitativemappingofthehemoglobin concentrationwithinthecells.Theaveragefluorescencelifetimesofuninfectedcohortswas270630ps(mean6SD;N=45). In the cytoplasm of infected cells the fluorescence lifetime of calcein ranged from 290620ps for cells with ring stage parasites to 590613ps and 1050660ps for cells with young trophozoites and late stage trophozoite/ early schizonts, respectively.Thiswasequivalenttoreductionsinhemoglobinconcentrationspanningtherangefrom7.3to2.3mM,inline with the model pred