Cytokine Analysis in Leukemia and Minimal Residual Disease (MRD) Detection Through Flow Cytometry

In clinical practice, the analysis of intracellular cytokines holds paramount importance for the effective treatment of various diseases. Flow cytometry (FCM) stands out as a highly efficient technique, leveraging specific fluorescence-labeled antibodies to detect intracellular cytokines. The fundamental steps of this assay encompass cell collection, fixation, permeabilization, blocking, intracellular staining, and subsequent analysis by FCM. Notably, FCM not only identifies cytokine-producing cells but also provides a platform for the assessment of the biological functions of these critical signaling molecules. This chapter presents a modified methodology aimed at enhancing the specificity of intracellular cytokine staining through FCM. An emphasis is placed on exploring the effects of key variables, including sample characteristics, temperature conditions, buffer formulations, and nuances associated with data acquisition and analysis. Beyond the methodological details, this chapter underscores the broader applications of intracellular cytokine analysis using FCM across diverse fields, including immunology, infectious diseases, and cancer research. Future perspectives shed light on emerging technologies, contributing to the ongoing evolution of intracellular cytokine analysis methodologies. In summary, this chapter provides a concise yet comprehensive guide to a modified approach for detecting specific intracellular cytokine staining using FCM. By emphasizing the effects of critical variables, this chapter equips researchers and clinicians with valuable insights to enhance the precision and reliability of intracellular cytokine analysis in diverse clinical and research settings.