Two-photon laser scanning fluorescence microscopy using photonic crystal fiber

Two-photon laser scanning fluorescence microscopy using photonic crystal fiber

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1、JournalofBiomedicalOptics9(5),922±927(September/October2004)Two-photonlaserscanning¯uorescencemicroscopyusingphotoniccrystal®berGailMcConnellAbstract.WereporttheapplicationofasimpleyetpowerfulmodularUniversityofStrathclydepulsecompressionsystembasedonphotoniccrystal®bersthati

2、m-CentreforBiophotonicsprovesonincumbenttwo-photonlaserscanning¯uorescencemicros-SIBS,27TaylorStreetGlasgowG40NR,Scotlandcopytechniques.Thissystemprovidesmorethanasevenfoldincreasein¯uorescenceyieldwhencomparedwithacommercialtwo-photonErlingRiismicroscopysystem.Fromthis,weinf

3、erpulsesofIRradiatonoflessUniversityofStrathclydethan35fsdurationreachingthesample.©2004SocietyofPhoto-OpticalIn-DepartmentofPhysicsstrumentationEngineers.[DOI:10.1117/1.1778734]JohnAndersonBuildingGlasgowG40NG,ScotlandKeywords:multiphoton;microscopy;¯uorescence;ultrafast;non

4、linear.E-mail:g.mcconnell@strath.ac.ukPaper03110receivedSep.3,2003;revisedmanuscriptreceivedJan.8,2004,andFeb.5,2004;acceptedforpublicationFeb.16,2004.1Introductionmadetoprecompensatefordispersionbyincludingprismsinahome-builtlaserresonator,deliveringpulsesof15.5fsMultiphoton

5、laserscanningmicroscopy~MPLSM!isawell-3establishedtoolfordeepandliveimagingofbiologicalduration,butthisapproachisin¯exiblewhenusedwithal-tissue.1Twotypesoflasersourcessuitedtotwo-photonexci-ternativecommercialresonators.tationexist,namely,continuouswave~cw!andultra-short-Toso

6、lvethisproblem,wedevelopedaneffectivepulsepulsed~USP!lasers,thelatterdeliveringpulsestypicallyofcompressiontechniquebasedonphotoniccrystal®ber~PCF!.4subpicoseconddurationeveryfewnanoseconds.TheresultingPreviousstudiesbyOuzounovetal.demonstratedthatul-peakpowerfromUSPlaserscan

7、easilyreachthekilowatttrashortpulsescanpropagatethroughPCF,butintheirinves-level,whileretainingamodestaveragepowerlevel.Theverytigationtheemittedpulsesarestretchedfrom100to140fs.highpeakpowersgeneratedbyUSPlasersourcesareidealforWereportonasystemthatcompressesthepulseduration

8、and,multiphotonapplicationswhenfocusedusinganobjectiveusingaconventi

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