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1、SCI.MAR.,64(2):183-195SCIENTIAMARINA2000AQUATICFLOWCYTOMETRY:ACHIEVEMENTSANDPROSPECTS.M.RECKERMANNandF.COLIJN(eds.)Probingthephysiologicalstateofphytoplanktonatthesingle-celllevel*FRANKJ.JOCHEMTheUniversityofTexasatAustin,MarineScienceInstitute,750ChannelViewDrive,PortAransas,TX783
2、73,USA.Fax:(361)7496777.E-Mail:frank@emcs.deSUMMARY:Probingthephysiologicalstateofphytoplanktonatthesingle-celllevelprovidesvaluableinsightinecologi-calstudiesaswellasinenvironmentalmonitoringofpollutionorUVimpacts.Thispaperreviewstherecentprogressinassessingthephysiologicalstateof
3、phytoplanktonwithflowcytometrybyinherentcellpropertiessuchascellsizeandchlorophyllautofluorescence,specificfluorescentdyes,andnewlydevelopedmolecularprobesandenzymesubstrates.Itisreportedhownitrogenandironlimitationaswellastheeffectofcopperpollutioncouldbederivedfromchangesincellin
4、herentproperties.EffectsofCuwerealsorecordedbymonitoringcellmembranepotentialsandesteraseactivity.Photo-syntheticcapacityofalgaewasassessedbychangesinchlorophyllfluorescencewiththeelectrontransportinhibitorDCMU,byacytometricadaptationofthepump-and-probeapproach,andmolecularprobesfo
5、rRubisco.Antibodieswerealsoappliedtomarknon-terminalstagesinthecellDNAreplicationcycle,todetectnon-proliferatingcells,toassessDNAdam-agecausedbyUV-Bradiationandtoquantifydiatomstickiness.Fluoresceindiacetateprovedusefultodiscriminatemeta-bolicallyactivefrominactivecellsandtoreveals
6、trategiesofdarksurvivalinalgae.TheactivityofalkalinephosphatasewasrecordedbyanewfluorigenicsubstrateELF,andpolyclonalantibodiesagainstnitratereductase(NR)providedmea-surementsoftheNRabundance.Anoutlookwillshowhowrecentdevelopmentsinmolecularprobesmightaffectthefutureanalysisofmarin
7、eecosystemsandtheircommunities.Keywords:flowcytometry,phytoplankton,physiology,molecularprobes,fluorescentdyes.INTRODUCTIONhighorganizationalcomplexity.Furthermore,theirsimpleorganizationmakesthedetectionofspecificIntimesofincreasingpublicawarenessandsensi-physiologicalreactionsand
8、conditionsmorefeasible.tiv
