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Fluorospot assay: methodological analysis.

Agnes Gazagne·Wolf Malkusch·Benoit Vingert·Wolf H Fridman·Eric Tartour
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Abstract

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.

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CytokinesEnzyme-Linked Immunosorbent AssayFluorescent DyesHumansIn Vitro TechniquesInterferon-gammaInterleukin-2T-Lymphocyte SubsetsJournal ArticleResearch Support, Non-U.S. Gov't

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Agnes Gazagne, Wolf Malkusch, Benoit Vingert, Wolf H Fridman, Eric Tartour. 2005. Fluorospot assay: methodological analysis.. https://doi.org/10.1385/1-59259-903-6%3A289

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