Summary Plants produce an estimated 350,000 metabolites, small molecules that are involved in everything from basic metabolism to species specific functions in signaling and defense against e.g. herbivores and drought. Their specific distributions across tissues is tightly linked to their biological function, however challenging to study due to their extreme chemical diversity. Mass Spectrometry Imaging (MSI) is developing as a key technology to visualize metabolic responses to environmental challenges and explore Nature's solutions for metabolite production and storage. The aim of the project is to optimize critical steps in sample preparation to achieve high-quality MSI results that will support research into crop plant resilience and biotechnological solutions for the green transition.