title: Visualizing and Tracking T Cell Motility In Vivo.
abstract: Advanced cellular tracking and imaging techniques allow the dynamic nature of immune responses to be studied in detail and in a physiological context. Here we describe two methods applying multiphoton laser scanning microscopy to the visualization and tracking of fluorescently labeled CD4+T cells and dendritic cells (DCs) within the complex lymph node (LN) environment. Ex vivo imaging of LNs allows the study of cell populations without the need for skilled surgical techniques while providing comparable data. While more technically demanding, intravital imaging of the popliteal LN allows aspects of T cell/DC responses to be studied in the context of an intact lymph and blood supply. We also describe methods to aid the acquisition of time series data suitable for cellular tracking, providing a quantitative approach to real-time analysis of DC and T cell LN responses.
authors.affiliation: Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, The University of Glasgow, Glasgow, G12 8TA, UK. robert.benson@glasgow.ac.uk.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Benson
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, Glasgow Biomedical Research Centre, University of Glasgow, B424, Sir Graeme Davies Building, 120 University Place, Glasgow, G12 8TA, UK.
authors.forename: James M
authors.initials: JM
authors.lastname: Brewer
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, Glasgow Biomedical Research Centre, Wellcome Trust Centre for Molecular Parasitology, University of Glasgow, B526, Sir Graeme Davies Building, 120 University Place, Glasgow, G12 8TA, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Garside
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002465:Cell Movement / Q000276:immunology*; D058948:Cell Tracking / Q000379:methods; D003713:Dendritic Cells / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods
keywords: Adoptive transfer; Dendritic cell; In vivo; Intravital imaging; Migration; Multiphoton; T cell