Qualitative and quantitative analysis of the immune synapse in the human system using imaging flow cytometry

• Verfasst von: Wabnitz, Guido H. [VerfasserIn]
• Verfasst von: Kirchgessner, Henning [VerfasserIn]
• Verfasst von: Samstag, Yvonne [VerfasserIn]
• Titel: Qualitative and quantitative analysis of the immune synapse in the human system using imaging flow cytometry
• Verf.angabe: Guido Wabnitz, Henning Kirchgessner, Yvonne Samstag
• E-Jahr: 2019
• Jahr: January 7, 2019
• Umfang: 1 Online-Ressource (1 Videodatei, 8:35 min)
• Illustrationen: farbig
• Fussnoten: Enthält auch Textversion ; Gesehen am 25.06.2021 ; Wissenschaftlicher Film. Deutschland. 2019
• Titel Quelle: Enthalten in: JoVE. Video journal
• Ort Quelle: [S.l.] : [s.n.], 2006
• Jahr Quelle: 2019
• Band/Heft Quelle: (2019), 143, Seite ?
• ISSN Quelle: 1940-087X
• DOI: doi:10.3791/55345
• Datenträger: Online-Ressource
• Dokumenttyp: Film
• Sprache: eng
• Sach-SW: Flow Cytometry
• Sach-SW: Humans
• Sach-SW: Immunological Synapses
• K10plus-PPN: 176128570X

Abstract

The immune synapse is the area of communication between T cells and antigen-presenting cells (APCs). T cells polarize surface receptors and proteins towards the immune synapse to assure a stable binding and signal exchange. Classical confocal, TIRF, or super-resolution microscopy have been used to study the immune synapse. Since these methods require manual image acquisition and time-consuming quantification, the imaging of rare events is challenging. Here, we describe a workflow that enables the morphological analysis of tens of thousands of cells. Immune synapses are induced between primary human T cells in pan-leukocyte preparations and Staphylococcus aureus enterotoxin B (SEB)-loaded Raji cells as APCs. Image acquisition is performed with imaging flow cytometry, also called In-Flow microscopy, which combines features of a flow cytometer and a fluorescence microscope. A complete gating strategy for identifying T cell/APC couples and analyzing the immune synapses is provided. As this workflow allows the analysis of immune synapses in unpurified pan-leukocyte preparations and hence requires only a small volume of blood (i.e., 1 mL), it can be applied to samples from patients. Importantly, several samples can be prepared, measured, and analyzed in parallel.