The classic view of metabolism is that enzymes are homogenously distributed across the cell and that they are regulated by mechanisms including competition, allosteric activation/inhibition, and feedback loops. However, over the past 40 years it has become clear that additional regulatory mechanisms bring enzymes together in time and space. Although enzyme assemblies have been observed across all domains of life, relatively little is known about the structure of such dynamic assemblies inside cells and the molecular mechanisms they use to regulate metabolism. In addition, there are few experiments that substantiate the activity of enzyme assemblies by directly monitoring their chemical reactions. We are leveraging state-of-the-art super-resolution optical photothermal infrared imaging to connect structure and function in enzyme assemblies associated with carbon metabolism in living cells. Overall, this work will uncover the importance of localized metabolism and additional complexity in carbon metabolism, which can later be expanded to other metabolic pathways.