title: A method for analyzing protein-protein interactions in the plasma membrane of live B cells by fluorescence resonance energy transfer imaging as acquired by total internal reflection fluorescence microscopy.
abstract: For more than a decade, fluorescence resonance energy transfer (FRET) imaging methods have been developed to study dynamic interactions between molecules at the nanometer scale in live cells. Here, we describe a protocol to measure FRET by the acceptor-sensitized emission method as detected by total internal reflection fluorescence (TIRF) imaging to study the interaction of appropriately labeled plasma membrane-associated molecules that regulate the earliest stages of antigen-mediated signaling in live B lymphocytes. This protocol can be adapted and applied to many cell types where there is an interest in understanding signal transduction mechanisms in live cells.
authors: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, USA.
authors: Hae Won
authors: HW
authors: Sohn
authors: Pavel
authors: P
authors: Tolar
authors: Joseph
authors: J
authors: Brzostowski
authors: Susan K
authors: SK
authors: Pierce
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism; D000941:Antigens / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D031541:Fluorescence Resonance Energy Transfer; D007091:Image Processing, Computer-Assisted; D008051:Lipid Bilayers / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D000941:Antigens; D008051:Lipid Bilayers; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases