Cytokines are major mediators for many cell signaling pathways, they play an important role in a various biological process including infection, inflammation, cancer, immune response, trauma and many diseases like Alzheimer. Considering the heterogeneity of Immune cells, cytokines detection is facing various challenging. Enzyme-linked immune spot (ELISpot) is the main tool to analyze cytokines, but it is able to analyze only one or two types of cytokines while many immune cells secrete more than 100 type of cytokines. In this work, a new microarray-based analytical tool is developed to assay the different immune cells secreted cytokines. After activating THP cells with LPS and incubating them for about 6 hrs, cytokines of IL1B, ILB, IL6, MIF, MMP3, and TNF-α have been analyzed using microarray-beads functionalized with antibodies to detect them. A decoding process has been performed using complementary DNA conjugated with Cy3 and Cy5 dyes to investigate what kind of cytokines has been actually secreted. This convenient method of making microbeads-based array is a reliable multiplex single-cell tool to evaluate different cytokines that secreted by immune cells.

Multiplex single-cell detection of cytokines by barcoded microarray method

256th National Meeting (2018)

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