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1、SCI.MAR.,64(2):165-181SCIENTIAMARINA2000AQUATICFLOWCYTOMETRY:ACHIEVEMENTSANDPROSPECTS.M.RECKERMANNandF.COLIJN(eds.)Immunoflowcytometryinmarinephytoplanktonresearch*L.PEPERZAK1,E.G.VRIELING2,B.SANDEE1andT.RUTTEN11NationalInstituteforCoastalandMarineMan
2、agement/RIKZ,P.O.Box8039,NL-4330EAMiddelburg,TheNetherlands.Fax(31)118651046;E-mail:L.Peperzak@rikz.rws.minvenw.nl2UniversityofGroningen,Dept.ofMarineBiology,P.O.Box14,NL-9750AAHaren,TheNetherlandsSUMMARY:Thedevelopmentsinthecombinationofflowcytometry
3、andimmunologyasatooltoidentify,countandexaminemarinephytoplanktoncellsarereviewed.Theconceptsofimmunologyandflowcytometryaredescribed.Adis-tinctionismadebetweenquantitativeandqualitativeimmunofluorescence.Quantitativeimmunofluorescence,theidentifi-cat
4、ionandenumerationofphytoplanktoncells,istheresearchareathathasadvancedrapidlyinthepastdecade,andisreviewedextensively.Keystepsofquantitativeimmunofluorescence,fixationandimmunolabelintensity,arediscussedinmoredetail.Qualitativeimmunofluorescenceisanew
5、,hardlyexploredbuthighlyinterestingdevelopmentinwhichquali-tative-physiological-variablesrelatedtoe.g.nutrientlimitationorprimaryproductionaremeasuredinindividualcellsinsteadofphytoplanktonpopulationsasawhole.Severalcombinationsofimmunologicalprobes,b
6、othforspeciesidentifi-cationandforphysiologicalmeasurements,areproposed.Aspecialcaseofqualitativeimmunofluorescenceisthemea-surementofphytoplanktontoxinsinsinglecellsfromnaturalpopulations.Itisanticipatedthatthefutureuseofsemicon-ductornanocrystalsor“
7、quantumdots”asfluorophoreswillgreatlyenhancesignaldetectioninflowcytometry,andhenceinbothquantitativeandqualitativeimmunofluorescenceapplications.Keywords:flowcytometry,immunology,marinephytoplankton,fluorescence,EuropeanOpticalPlanktonAnalyser(EurOPA
8、),physiology,antibodiesINTRODUCTIONprokaryotes(DahlandLaake,1982;Campbelletal.,1983),butsomeyearslatereukaryoticorganismsImmunologyandflowcytometryarebecomingfollowed(Hiroishetal.,1988;Shapiroetal.,1989).powerfultoolsinthestudyofphytoplanktone
