2025-W6: Demystifying Stable Isotope Labelling Metabolomics

Presenters:  Dr Bart Ghesquiere (KU Leuven)  Dr James MacRae (The Francis Crick Institute) Description: Metabolite profiling and targeted metabolomics approaches are now routinely applied by researchers wanting to understand the physiology of their cell, tissue, organism, or biofluid of choice. One of the largest challenges faced by metabolomics researchers in any study is relating changes in identified metabolites to their metabolic roles. Understanding this relationship is critical for understanding metabolic mechanisms in that biosystem. Stable isotope labelling has rapidly emerged as a powerful tool in unravelling the precise structure and activity of metabolic pathways and how these influence/are caused by the physiological state of the biosystem. These techniques can reveal subtle alterations in metabolic activity that would not be possible with unlabeled approaches. This depth of information can provide valuable insight into the mechanisms that underlie various physiological conditions and aberrant processes, including infection and disease. However, this technique is perhaps not used as widely as it might. This workshop will show how metabolic labelling studies are readily approachable and easily interpretable when performed correctly. We will show you how to design, run, and analyse stable isotope labelling experiments, and explain how these simple workflows are often sufficient for significant biological insight. New for 2025! Simple in vivo labelling and imaging approaches Workshop Objectives: Attendees will leave the workshop:  Knowing how to design a labelling project, including essentials to consider and potential pitfalls  Knowing the basics of analysis of labelling data, including awareness of some of the available LC-MS and GC-MS software  Understanding how to interpret data, including the importance of mass isotope distributions Learning Outcomes: Attendees will learn:  That stable isotope labelling projects are easy to design, perform, and interpret  That labelling experiments can bring intricate detail to many metabolomics projects, whether untargeted or targeted, or polar or lipid-focused  Key principles for in vitro and in vivo labelling studies, combining labelling with mass spectrometry imaging, and multi-isotope approaches