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AbstractIntracellular organelles have a particular morphological signature that can only be appreciated by ultrastructural analysis at the electron microscopy level. Optical imaging and associated methodologies allow to explore organelle localization and their dynamics at the cellular level. Deciphering the biogenesis and functions of lysosomes and lysosome-related organelles (LROs) and their dysfunctions requires their visualization and detailed characterization at high resolution by electron microscopy. Here, we provide detailed protocols for studying LROs by transmission electron microscopy. While conventional electron microscopy and its recent improvements is the method of choice to investigate organelle morphology, immunoelectron microscopy allows to localize organelle components and description of their molecular make up qualitatively and quantitatively.
CitationHurbain I, Romao M, Bergam P, Heiligenstein X, Raposo G (2017) Analyzing Lysosome-Related Organelles by Electron Microscopy. Lysosomes: 43–71. Available: http://dx.doi.org/10.1007/978-1-4939-6934-0_4.
SponsorsWe are grateful to Institut Curie and the BioImaging platform (PICT IBiSA), member of the France-BioImaging national research infrastructure (ANR-10-INSB-04) and CNRS. Research in our group is supported by the Fondation pour la Recherche Médical (Equipes FRM), Association de Recherche pour le Cancer (ARC), Indian French cooperation (CEFIPRA), Clarins and L’Oréal.
JournalMethods in Molecular Biology