Dietary sphingolipids are significant components of infant diets that have great potential to interact with beneficial gut microbes with active sphingolipid metabolism. Despite the prevalence of sphingolipid-producing bacteria in infant microbiomes, the ability of the gut microbiome to take up and transform dietary sphingolipids has not been fully explored. During this webinar I will discuss methodology my lab has developed to trace dietary sphingolipids into gut microbial metabolism using a combination of click chemistry, fluorescence activated cell sorting, 16S rRNA gene sequencing, and comparative metabolomics. With this approach we determined that sphingolipid-producing bacteria readily take up dietary sphingolipids. We also characterized the use of exogenous sphingolipids by other beneficial bacteria not previously known to interact with sphingolipids. Together this work opens up the need for further exploration of sphingolipids as important molecules in diet-host-microbe interactions.