Hyperpolarized Micro-NMR Platform for Sensitive Analysis of In Vitro Metabolic Flux in Living Cells

Metabolism represents an ensemble of cellular biochemical reactions, and thus metabolic analyses can shed light on the state of cells. Metabolic changes in response to external cues, such as drug treatment, for example, can be rapid and potentially an early indicator of therapeutic response. Unfortunately, conventional techniques to study metabolism, such as optical microscopy or mass spectrometry, have functional limitations in specificity and sensitivity. To address this technical need, we developed a sensitive analytical tool based on nuclear magnetic resonance (NMR) technology, termed hyperpolarized micro-NMR, that enables rapid quantification of multiple metabolic fluxes in a small number of cells, down to 10,000 cells, nondestructively. This analytical capability was achieved by miniaturization of an NMR detection coil along with hyperpolarization of endogenous metabolites. Using this tool, we were able to quantify pyruvate-to-lactate flux in cancer stem cells nondestructively within 2 min, which has not been possible with other techniques. With further optimization, we envision that this novel device could be a powerful analytical platform for sensitive analysis of metabolism in mass-limited samples.