@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.}
}