title: Analyzing Actin Dynamics at the Immunological Synapse.
abstract: T cell signaling is inextricably linked to actin cytoskeletal dynamics at the immunological synapse (IS). This process can be imaged in living T cells expressing GFP actin or fluorescent F-actin binding proteins. Because of its planar nature, the IS provides a unique opportunity to image events as they happen, monitoring changes in actin retrograde flow in T cells interacting with different stimulatory surfaces or after pharmacological treatments. Here, we described the imaging methods and analytical procedures used to measure actin velocity across the IS in T cells spreading on planar stimulatory surfaces.
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, 3615 Civic Center Blvd, ARC 816D, Philadelphia, PA, 19104, USA.|Perelman School of Medicine of the University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Katarzyna I
authors.initials: KI
authors.lastname: Jankowska
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, 3615 Civic Center Blvd, ARC 816D, Philadelphia, PA, 19104, USA. jburkhar@mail.med.upenn.edu.|Perelman School of Medicine of the University of Pennsylvania, Philadelphia, PA, USA. jburkhar@mail.med.upenn.edu.
authors.forename: Janis K
authors.initials: JK
authors.lastname: Burkhardt
mesh_terms: D000199:Actins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D054992:Immunological Synapses / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019169:Jurkat Cells; D051379:Mice; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism
chemical_list: D000199:Actins; D049452:Green Fluorescent Proteins
keywords: Actin; Cytoskeleton; Immunological synapse; Integrin; Kymograph; Live cell imaging; Mobile ligands; Planar lipid bilayer; Spinning disk; T-cells