A Tandem CytometryPlatform for Single-Cell Analysisof Protein and Metabolites

Multiomics analysis at the single-cell level is essential for both fundamental research and clinical applications, with proteomics and metabolomics being particularly crucial for providing insights into cellular states and functions. The state of the art flow cytometry has shown great potential in identifying cellular proteins, while emerging metabolite mass spectrometry cytometry techniques address metabolite detection. Herein, we propose a tandem platform that integrates fluorescence flow cytometry with electrospray ionization mass spectrometry for one-step single-cell analysis of protein and metabolites. An algorithm was established to correlate multidimensional information in individual cells, with additional data processing modules designed to ensure accuracy and facilitate further analysis. The tandem cytometry platform demonstrated efficacy in profiling breast cancer cells, particularly under hypoxic conditions, revealing metabolic shifts with decreased glutathione and increased l-glutamine levels, indicative of hypoxia-inducible factor activity. This platform introduces a powerful analytical capability that promises to elevate the precision of cell-based diagnostics and therapeutic strategies.