title: Secondary metabolite localization by autofluorescence in living plant cells.
abstract: Autofluorescent molecules are abundant in plant cells and spectral images offer means for analyzing their spectra, yielding information on their accumulation and function. Based on their fluorescence characteristics, an imaging approach using multiphoton microscopy was designed to assess localization of the endogenous fluorophores in living plant cells. This method, which requires no previous treatment, provides an effective experimental tool for discriminating between multiple naturally-occurring fluorophores in living-tissues. Combined with advanced Linear Unmixing, the spectral analysis extends the possibilities and enables the simultaneous detection of fluorescent molecules reliably separating overlapping emission spectra. However, as with any technology, the possibility for artifactual results does exist. This methodological article presents an overview of the applications of tissular and intra-cellular localization of these intrinsic fluorophores in leaves and fruits (here for coffee and vanilla). This method will provide new opportunities for studying cellular environments and the behavior of endogenous fluorophores in the intracellular environment.
authors: Institut des Sciences de l'Evolution Montpellier ISE-M, Université Montpellier, CNRS, IRD, EPHE, CC 065, Place Eugène Bataillon, 34095 Montpellier, France. pascale.talamond@ird.fr.
authors: Pascale
authors: P
authors: Talamond
authors: Histocytology and Plant Cell Imaging platform PHIV, UMR AGAP (CIRAD, INRA, SupAgro)-UMR B&PMP (INRA, CNRS, SupAgro, Montpellier University), 34095 Montpellier, France. verdeil@cirad.fr.
authors: Jean-Luc
authors: JL
authors: Verdeil
authors: Histocytology and Plant Cell Imaging platform PHIV, UMR AGAP (CIRAD, INRA, SupAgro)-UMR B&PMP (INRA, CNRS, SupAgro, Montpellier University), 34095 Montpellier, France. genevieve.conejero@supagro.inra.fr.
authors: Geneviève
authors: G
authors: Conéjéro
mesh_terms: D040503:Coffea / Q000737:chemistry*; D005638:Fruit / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D010636:Phenols / Q000032:analysis; D059828:Plant Cells / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D064210:Secondary Metabolism; D031669:Vanilla / Q000737:chemistry*
chemical_list: D010636:Phenols