Recent advances in immunometabolism have demonstrated the importance of metabolic reprogramming in supporting the activation and effector functions of immune cells. Specifically, metabolic enzymes and metabolites have been shown to perform moonlighting functions, such as regulation of inflammation, in addition to their traditional metabolic roles. However, to fully understand their roles in orchestrating an inflammatory response in immune cells, specific techniques are required to assess the metabolic parameters of immune cells, such as the abundance and fate of metabolites, the activity, and localization of metabolic enzymes, as well as the direction of metabolic fluxes and the activity level of different metabolic pathways. In this Methods Collection, we are looking to feature innovative techniques that profile the metabolism of immune cells, including, but not limited to, seahorse analyzer assays of specific immune cell types, targeted and untargeted metabolomics, enzymatic cycling assays, or genetically encoded metabolic sensors. Novel assays that characterize the metabolic profiles of immune cell populations in vivo also fall within the scope of the topic.