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1、SCI.MAR.,64(2):197-224SCIENTIAMARINA2000AQUATICFLOWCYTOMETRY:ACHIEVEMENTSANDPROSPECTS.M.RECKERMANNandF.COLIJN(eds.)Usingflowcytometryforcountingnaturalplanktonicbacteriaandunderstandingthestructureofplanktonicbacterialcommunities*JOSEPM.GASOL1andPAULA.DELGIORGIO21DepartamentdeBiologiaMar
2、inaiOceanografia,InstitutdeCiènciesdelMar,CSIC.PasseigJoandeBorbós/n.E08039Barcelona,Catalunya.Spain.Phone:(3493)2216416,Fax:(3493)2217340.E-mail:pepgasol@icm.csic.es2HornPointLaboratory.UniversityofMarylandCenterforEnvironmentalSciencePO.Box775Cambridge,MD21613USASUMMARY:Flowcytometryis
3、rapidlybecomingaroutinemethodologyinaquaticmicrobialecology.Thecombinationofsimpletousebench-topflowcytometersandhighlyfluorescentnucleicacidstainsallowsfastandeasydeterminationofmicrobeabundanceintheplanktonoflakesandoceans.Thedifferentdyesandprotocolsusedtostainandcountplankton-icbacte
4、riaaswellastheequipmentinusearereviewed,withspecialattentiontosomeoftheproblemsencounteredindailyroutinepracticesuchasfixation,stainingandabsolutecounting.Oneofthemainadvantagesofflowcytometryoverepi-fluorescencemicroscopyistheabilitytoobtaincell-specificmeasurementsinlargenumbersofcells
5、withlimitedeffort.Wediscusshowthischaracteristichasbeenusedfordifferentiatingphotosyntheticfromnon-photosyntheticprokaryotes,formeasuringbacterialcellsizeandnucleicacidcontent,andforestimatingtherelativeactivityandphysiologicalstateofeachcell.Wealsodescribehowsomeoftheflowcytometricallyo
6、btaineddatacanbeusedtocharacterizetheroleofmicrobesoncarboncyclingintheaquaticenvironmentandweprospectthelikelyavenuesofprogressinthestudyofplanktonicprokaryotesthroughtheuseofflowcytometry.Keywords:heterotrophicandphototrophicbacteria,flowcytometry,countingbacteria,activebacteria.INTROD
7、UCTIONetal.,1997)thatattimescanbehigherthan,oratleastsimilarto,thatofprimaryproducers(Lietal.,Bacteriaareimportantcomponentsofplanktonic1992,Fuhrmanetal.,1989;Simonetal.,1992).foodwebs:autotrophicbacteriacandominatetheBacterialcontributiontoplanktonicheterotrophicprimaryp
