In fact, the Single-probe SCMS techniques are primarily used for live mammalian cell studies. Sun et al. compared the metabolic profiles of cancer stem cells (CSCs) with non-stem cancer cells (NSCCs). This study revealed distinct metabolic features in CSCs, particularly elevated tricarboxylic acid (TCA) cycle metabolites and unsaturated lipids. These metabolites, regulated by key enzymes such as stearoyl-CoA desaturase-1 (SCD1), were linked to the stemness and drug resistance of CSCs. This work demonstrated the ability of SCMS to reveal unique metabolic pathways in CSCs, which could serve as potential therapeutic targets. This application of SCMS illustrated its effectiveness in real-time, live-cell metabolomics, advancing our understanding of cancer biology at the single-cell level.