Cytokine Flow Cytometry

Cytokine flow cytometry (CFC) is a general term that applies to flow cytometric analysis of cells using anticytokine antibodies as markers of activation. The most common version of this technique is the intracellular staining of cytokines in cells that have been fixed and permeabilized after short-term in vitro activation. When used with specific antigens, this technique allows for the quantitation of rare populations of antigen-specific T cells. In this chapter, specific methodology for such intracellular staining is elaborated, with emphasis on the effects of variables such as sample type, antigens, activation conditions, sample processing, and data acquisition and analysis.