Fluorospot assay: methodological analysis.
The conventional enzyme-linked immunospot (ELISPOT) technique detects only one secreted cytokine, which constitutes a major pitfall for the accurate characterization of the various T-cell subpopulations. We have therefore developed a fluorospot assay, which is a modification of the ELISPOT and is based on the use of multiple fluorescent-labeled anticytokines detection antibodies. A special automated ELISPOT reader consisting of a light microscope with incident fluorescence illumination and an integrating digital color camera has been adapted for this technique. This technique has been applied for the analysis of subpopulations of T-cells and polarized antigen-specific T-cells.
Gazagne
Agnes
A
Unité d'Immunologie Biologique, Hôpital Européen Georges Pompidou, Université René Descartes, Paris, France.
Malkusch
Wolf
W
Vingert
Benoit
B
Fridman
Wolf H
WH
Tartour
Eric
E
D016207:Cytokines / Q000032:analysis* / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis; D007376:Interleukin-2 / Q000032:analysis / Q000096:biosynthesis; D016176:T-Lymphocyte Subsets / Q000276:immunology
D016207:Cytokines; D005456:Fluorescent Dyes; D007376:Interleukin-2; D007371:Interferon-gamma