Our knowledge of the multiplicity and functions of cytokines has expanded greatly during the last 10 years. However, the large number and low concentrations of cytokines that are expressed in a pathogenic condition often preclude determining the presence and function of each cytokine during disease pathogenesis (1)(2) . The FlowMetrixTM system has been developed as a highly sensitive, multiplexed assay platform that provides a means to determine qualitative and quantitative measurements of multiple analytes simultaneously. The system utilizes polystyrene microspheres as a vehicle for different molecular reactions to occur. Differential staining of these microspheres with orange-emitting and red-emitting fluorochromes produces an array of 64 individually addressable populations of microspheres (3) . A green-emitting fluorochrome is coupled to detection molecules to measure each molecular reaction. A graphical representation of the classification of each microsphere set and the subsequent measurement of green fluorescence associated with each set is illustrated in Fig. 1⇓ A. The use of a standard flow cytometer and its power to analyze the concentrations of three different fluorochromes on individual particles allow the simultaneous quantification of up to 64 different cytokines or chemokines in a single tissue or fluid sample in real time. We have assembled a number of different multiplexed cytokine panels to analyze proinflammatory cytokines and …
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