title: Using Confocal Microscopy to Investigate Intracellular Trafficking of Toll-Like Receptors.
abstract: Toll-like receptors (TLR) survey the extracellular space, cytoplasm, and endosomal compartments for signs of infection or tissue injury. Over the past decade, it has become evident that TLR activation and signal transduction can be regulated by subcellular compartmentalization of both the receptors and their downstream signaling components. Immunofluorescence and/or overexpression of fluorescently "tagged"' proteins teamed with confocal microscopy presents a powerful technique for studying the spatial organization of TLRs, their signaling mediators, and the dynamic processes they activate. This chapter details the common methods for determining the subcellular location of TLRs in both live and fixed cells.
authors.affiliation: Centre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, Norway.|Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
authors.forename: Harald
authors.initials: H
authors.lastname: Husebye
authors.affiliation: Department of Clinical Medicine, School of Medicine, Trinity College Dublin, Dublin 2, Ireland. sarah.doyle@tcd.ie.|The National Children's Research Centre, Our Lady's Children's Hospital Crumlin, Dublin 12, Ireland. sarah.doyle@tcd.ie.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Doyle
mesh_terms: D000818:Animals; D002460:Cell Line; D015870:Gene Expression; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D007963:Leukocytes, Mononuclear; D008024:Ligands; D018613:Microscopy, Confocal / Q000379:methods; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism*
chemical_list: D008024:Ligands; D011993:Recombinant Fusion Proteins; D051193:Toll-Like Receptors
keywords: Confocal microscopy; Fluorophore; GFP; Immunocytochemistry; Immunofluorescence; Subcellular localization; TLR trafficking