A Label-Free Dual-Mode Flow Cytometry for Single-Cell Physicochemical Property Analysis

Label-free flow cytometry is a powerful technique for cellular property characterization. So far, there has been no cytometry reported to obtain both physical and chemical properties at the single-cell level. Herein, we propose a label-free dual-mode flow cytometry that integrates impedance cytometry and mass cytometry. Single cells flowing through the impedance cytometry are individually characterized with electrical (cytoplasm conductivity, and specific membrane capacitance) and mechanical (Young's modulus, and fluidity) parameters. They are subsequently ionized by downstream on-chip electrospray and characterized with chemical components (42 types of metabolites and lipids) via mass spectrometry analysis. We conducted a pilot test of three different human leukemia cell lines (THP-1, Jurkat, and HL-60 cells) to obtain their physicochemical properties. We found that using multimodal properties for cell typing achieved higher accuracy (∼11% increase) than single-mode properties. More interestingly, property correlation analysis indicates that lipid PC(P-36:0) may be a common component contributing to regulating the electrical properties (i.e., specific membrane capacitance) for all three cell lines, but the major component for each cell line may be different (e.g., PE(P-36:4) for HL-60 cytoplasm conductivity). For some exceptional cells (5% outliers), cell conductivity exhibits a very high correlation coefficient (-0.83) with PC(38:2). These observations, though preliminary, only became available by our proposed dual-mode flow cytometry, and open a new perspective for investigating the molecular mechanisms underpinning cellular biophysics at the single-cell level. We envision that this work would foster a lot of research opportunities in multimodal cell characterization and provide new clues for molecular cell biology.