Understanding metabolic fluxes is crucial for elucidating the biochemical mechanisms underlying diseases, drug responses, and genetic mutations. Tracer metabolomics offers a unique window into these dynamic processes by tracking labelled substrates through metabolic pathways. However, interpreting tracer metabolomics data is technically challenging, as it requires the integrated analysis of metabolite levels, isotopic tracer incorporation patterns, and their specific localization within metabolic pathways. This three-day course is designed for researchers who are interested in learning the basics of tracer metabolomics. Participants will gain foundational knowledge in the concepts needed to set up a tracer metabolomics experiment and to understand and interpret the results of a tracer metabolomics analysis.