title: Automated nanoscale flow cytometry for assessing protein-protein interactions.
abstract: Despite their importance for signalling events, protein-protein interactions cannot easily be analyzed on a single cell level. We developed a robust automated FRET measurement system implemented on a commercial flow cytometer allowing for rapid profiling of molecular associations in living cells. We used this method to measure the most proximal signaling events on human T lymphocyte activation, which preceded calcium influx, and could automatically detect T cell receptor/CD3 complex clustering defects in immunocompromised patients. © 2016 International Society for Advancement of Cytometry.
authors: Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, Nordrhein Westfalen, 51429, Germany.
authors: Kerstin
authors: K
authors: von Kolontaj
authors: Institute of Innate Immunity, University Hospitals, University of Bonn, Sigmund-Freud-Str. 25, Bonn, 53127, Germany.
authors: Gabor L
authors: GL
authors: Horvath
authors: Institute of Innate Immunity, University Hospitals, University of Bonn, Sigmund-Freud-Str. 25, Bonn, 53127, Germany. eicke.latz@uni-bonn.de.|Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, 01605. eicke.latz@uni-bonn.de.|German Center for Neurodegenerative Diseases, Bonn, 53175, Germany. eicke.latz@uni-bonn.de.|Centre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, Norway. eicke.latz@uni-bonn.de.
authors: Eicke
authors: E
authors: Latz
authors: Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, Nordrhein Westfalen, 51429, Germany. Martin@miltenyibiotec.de.
authors: Martin
authors: M
authors: Büscher
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008213:Lymphocyte Activation / Q000235:genetics / Q000276:immunology; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps / Q000235:genetics*; D059010:Single-Cell Analysis; D013601:T-Lymphocytes / Q000276:immunology
keywords: FRET; T cells; automatic data analysis; detection of immunodeficiencies; flow cytometry