Single Cell Analysis of the Proinflammatory Responses of Mast Cell by a Real Time Secretion Assay

A real-time secretion monitoring assay from living single-cells was realized by total internal reflection fluorescence imaging with an optically optimized microwell array chip fabricated of an amorphous fluoropolymer, CYTOP on the basis of a fluorescence sandwich immunoassay. After individual mast cells were stimulated with lipopolysaccharide in the microwell array, their proinflammatory responses were monitored as cytokine secretion along time. mRNA levels of cytokines by single-cell quantitative PCR was also measured after the secretion assay. The results showed that each of stimulated mast cells is quantitatively and qualitatively distinguishable from each other in terms of cytokine secretion and their mRNA profile.