Fig. 1 |. A general immunometabolism workflow.

The discovery phase of an immunometabolism project can originate from a multitude of techniques. Extracellular flux analysis (EFA) assays are useful to build a foundation to determine whether metabolic differences are involved in your phenotype of interest. These are especially effective when paired with normalization imaging technology. Next, metabolic phenotypes can be pursued with additional methods. Flow cytometric assays with metabolic dyes and lactate assays are user friendly and provide easily accessible options for most labs. In some instances, these assays also provide single-cell information. Some users may find flow cytometry-based methods more accessible for preliminary validations of a metabolic phenotype, followed by phenotype assessment with EFA. Next, metabolites may be assayed directly using targeted metabolomics. Flux measurements paired with metabolomics provide a detailed overview for metabolomics research and offer a wealth of mechanistic details underlying a metabolic phenotype. Finally, analysis of the phenotype in vivo using a genetic approach provides further support for the phenotype. CyTOF, cytometry by time of flight; log₂FC, log₂ fold change; RNA-seq, RNA sequencing.