Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale

















Transverse project I: Cell imaging of glycomolecules

Principal investigator : A. Driouich, Professor and S. Bernard, Engineer.
Participants : M-L. Follet-Gueye, Associate professor ; C. Burel, Technician ; G. Lucas, Technician.

Cell imaging is one of our main approaches for understanding the functional properties of plant glycomolecules in relation with development and defense. During the last decade, we have been able to develop and use various imaging technologies to address key questions related to the biosynthesis and function of cell wall polysaccharides and glycoproteins in a number of plant species (Dardelle et al., 2010; Driouich et al., 2012; Follet Gueye et al., 2012; Cannesan et al., 2012; Plancot et al., 2013; Leroux et al., 2015). These technologies include, light/fluorescence microscopy, confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM) coupled to high pressure freezing (HPF) for sample preparation and immunocytochemistry for glyco-epitopes localization. In the current proposal, we will continue to improve and employ our imaging strategies to address various questions related to the spatial and functional organization of plant glycomolecules. We will optimize the TEM/HPF technique that we have previously developed for land plants, to study microalgae glycomolecules (Topic III). We will introduce the use of a new imaging technique called “stimulated emission depletion” (STED) microscopy which provides a much higher resolution than classical confocal microscopy (Topics I - III). We will also develop a CLEM (correlative Light Electron Microscopy) technique that combines confocal laser scanning microscopy, high pressure freezing and high resolution TEM microscopy (Giepmans, 2008; McDonald, 2009) to image subcellular processes related to the biosynthesis and localization of cell wall glycomolecules as presented in different lab projects (see above, Topic I and II).
References : Giepmans (2008) Histochem. Cell. Biol. 130, 211-217; McDonald (2009) J. Microscopy 235, 273-281.