title: Enzyme-Linked ImmunoSpot (ELISpot) for Single-Cell Analysis.
abstract: The ELISpot, a heterogeneous immunoassay, is widely used for detection of low abundant analytes. It is a reliable and robust assay to monitor responses of the immune system at the single-cell level by capturing secreted molecules of interest with specific, membrane-bound antibodies. Those molecules are then made visible by a cascade of ELISA-related development steps. The final results are distinct spots on the membrane as an imprint of the cell secreting the captured molecules, not only allowing their quantification but also providing insight on the kinetics and strength of secretion. This chapter describes the optimized protocol steps of the ELISpot technique, important improvements and tools available for the community, and the current expansion of the technique into polyfunctional cell analysis.
authors.affiliation: ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ, 07024, USA. sylvia@zellnet.com.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Janetzki
authors.affiliation: ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ, 07024, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Rabin
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology; D007376:Interleukin-2 / Q000032:analysis / Q000276:immunology; D059010:Single-Cell Analysis / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D007376:Interleukin-2; D007371:Interferon-gamma
keywords: B cell assays; ELISpot; FluoroSpot; Immune monitoring; T cell assays