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1、MicrotubuleSegmentationand3DReconstructioninTotalInternalReflectionFluorescenceMicroscopy(TIRFM)StathisHadjidemetriou,DerekToomre,andJamesS.Duncan,YaleUniversity,DepartmentofBiomedicalEngineeringandDiagnosticRadiology,DepartmentofCellBiology,NewHaven,CT06520stathis@noodle.med.yale.ed
2、u,derek.toomre@yale.edu,duncan@noodle.med.yale.eduAbstractTheinteractionofthemicrotubulesandtheirassemblywiththecellmembraneplaysnumerouscriticalrolesinacell.Forexample,itdirectsneurotransmittervesiclestosynapsesandmodulatesmembraneadhesionspivotalforcellmovementandmitosis.Abnormalf
3、unctionoftheassemblyisinvolvedinneurodegenerativediseasesandcancer.AtechniquethatenablesthedirectobservationofmicrotubulefunctionnearthecellmembraneisTIRFmicroscopy.Todatemostimageanddataanalysisofthemicrotubuleassemblyhasbeendonebyvisualinspectionandmanualtracing.Inthisworkwehavede
4、velopedamethodtoautomaticallyextractmicrotubulesfromTIRFmicroscopyimagessoastoenableamoreextensiveandhigherthroughputquantitativestudyofthemicrotubuleassembly.Themicrotubulesarefirstsegmentedin2Dintermsofconsecutivesegments.ThesegmentsarestreamlinesofHamilton-Jocobicostmaps.Subsequen
5、tly,thefluorescenceisusedtoimplementalimitedreconstructionofthemicrotubulesin3Dspace.Last,weshowexamplesofourmethodappliedtophantomdataaswellasrealTIRFmicroscopyimagesfromfluorescentlytaggedlivingcells.Keywords:Totalinternalreflectionfluorescencemicroscopy,microtubuleassembly,segmentati
6、onoftubularstructures,Hamilton-Jacobiequation,anisotropicfastmarching.1Introduction1.1SignificanceofMicrotubulesMicrotubulesarebiopolymersofthecytoplasm.Theyarecomposedoftubulinandhaveadiameterof≈25nm[1].Inmammalianinterphasecellsoneendisanchoredatanorganellecalledthemicrotubuleorgan
7、izingcenter(MTOC),whichispositionedinthecellcenter.HundredsofmicrotubulesemergeradiallyfromtheMTOClikespokesoutofawheel.Theycancomeincloseproximitytothecellcortexanddynamicallyprobethecellperipherybyrapidlygrowingandshrinking[1].Inadditiontoprovidingstructuralstability,theyserveasde
8、liverytracksforthea
