@InProceedings{Supelec939,
author = {Mathieu Lauffer and Frédéric Genty and Samuel Margueron and Jean-Luc Collette and Annie Levey and Yves Houzelle and Jean-Claude Pihan},
title = {Fluorescence imaging of plant pigments for automatical recognition of aquatic organisms},
year = {2014},
booktitle = {Proceedings of the XII Conference on Optical Chemical Sensors and Biosensors},
pages = {FS-29},
address = {Athens (Grèce)},
abstract = {Currentlyspecialistsusuallyusemorphologicalparameterstoidentif yaquaticplants.Manystudieswererealizedonspectroscopicanalysiso fphytoplanktonpigments[1,2].Ourapproachisdifferent,wehavestudi edopticalpropertiesofphytoplanktoninordertodevelopafluorescenc eimaging.Thissystemallowstoidentifyseveralpigmentsinorganisms. Thegoalistodeterminethefamilyofmicroormacroalgaeaccordingtopig mentscomposition.Introduction:Basedonfluorescencemicroscopy,we developedanimagingsystemwhichhighlightschlorophyllswithdoublef ilter(excitationfiltersandemissionfilters).Afterspectroscopics tudyofchlorophylls(absorptionandphotoluminescenceinducedbylase r),wechoseexcitationfilterwithabsorptionbands(SoretbandandQyba nd)andemissionfilterwithemissionbandsofeachchlorophyll[3].Reco gnition of blue algae (cyanobacteria): Recognition of green algae (chlorophycea): TheCyanobacteriaweselectedwasAnabaenaflosaquae,afilamentousorg anismwithsphericalcells.Werealizedavisiblepicture.Thenfluoresc encepicturesweremadeforchlorophyll-a,chlorophyll- bandchlorophyll- d.Segmentation«watershed»wasappliedtovisiblepictureinordertoid entifydifferentareas.Wecalculatedmeanvectorsineachareaforfluor escencepictures(3picturesfor3wavelengthsor3chlorophylls).Thens egmentation«k- means»wereappliedtofluorescencepictureswithK=10(numberofcluste rs)andh=0,01(thresholddistanceforregionmerging).TheChlorophyce aweselectedwasCladophorasp,afilamentousorganismramifiedwithcyl indricalcells.Werealizedavisiblepicture.Thenfluorescencepictur esweremadeforchlorophyll-a,chlorophyll-bandchlorophyll- d.Segmentation«watershed»wasappliedtovisiblepictureinordertoid entifydifferentareas.Wecalculatedmeanvectorsineachareaforfluor escencepictures(3picturesfor3wavelengthsor3chlorophylls).Thens egmentation«k- means»wereappliedtofluorescencepictureswithK=10(numberofcluste rs)andh=0,003(thresholddistanceforregionmerging).Conclusion:Re ferences:Visible picture of Anabaena flos aquaeFluorescence of chlorophyll-aFluorescence of chlorophyll-bFluorescence of chlorophyll-dFluorescence of chlorophyll-dFluorescence of chlorophyll-bFluorescence of chlorophyll-aVisible picture of Cladophora spSegmentation «watershed» on visible pictureSegmentation «k-means» on fluorescence picturesSegmentation «watershed» on visible pictureSegmentation «k-means» on fluorescence pictures3D-map of the repartition of fluorescence intensity at 3 wavelengths corresponding to the 3 chlorophylls (chlorophyll-a, chlorophyll-b and chlorophyll-d) in Anabaena flos aquae3D-map of the repartition of fluorescence intensity at 3 wavelengths corresponding to the 3 chlorophylls (chlorophyll-a, chlorophyll-b and chlorophyll-d) in Cladophora sp[1]L.Poryvkina,S.Babichenko,A.Leeben,Analysisofphytoplankton pigmentsbyexcitationspectraoffluorescence,EARSeL-SIG- WorkshopLIDAR,InstituteofEcology/LDI,Tallinn,Estonia,224- 232(2000) [2]N.T.An,andH.T.Du."Studiesonphytoplanktonpigments:chlorophyl l,totalcarotenoidsanddegradationproductsinVietnamesewaters."Pr oceedingoftheSEAFDECSeminaronFisheryResourcesintheSouthChinaSe aAreaIV:VietnameseWater,SoutheastAsianFisheriesDevelopmentCent er,Thailand.2000.[3]Grimm,B.(Ed.). (2007).Chlorophyllsandbacteriochlorophylls:biochemistry,biophy sics,functionsandapplications(Vol.25).Springer.Withfluorescenc eimaging,wedevelopedanoriginalsystemwhichisabletorecognizethef amilyofmicroormacroalgaeandshouldbeveryhelpfulforspecificrecog nization.Opticalsetupcombiningexcitationfiltersandemissionfilt ersmakesitpossibletobeveryspecific.Imagingandstatisticalproces singbysegmentation(«watershed»and«k- means»)havebeendeveloppedinordertocreateadatabase.}
}