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[Preprint]. 2024 May 12:2024.05.08.593179. [Version 2] doi: 10.1101/2024.05.08.593179

SCO-pH: Microfluidic dynamic phenotyping platform for high-throughput screening of single cell acidification

Hyejoong Jeong, Emilia A Leyes Porello, Jean Gabriel Rosario, Da Kuang, Syung Hun Han, Jai-Yoon Sul, Bomyi Lim, Daeyeon Lee, Junhyong Kim
PMCID: PMC11100697  PMID: 38766224

Abstract

Studies on the dynamics of single cell phenotyping have been hampered by the lack of quantitative high-throughput metabolism assays. Extracellular acidification, a prominent phenotype, yields significant insights into cellular metabolism, including tumorigenicity. Here, we develop a versatile microfluidic system for single cell optical pH analysis (SCO-pH), which compartmentalizes single cells in 140-pL droplets and immobilizes approximately 40,000 droplets in a two-dimensional array for temporal extracellular pH analysis. SCO-pH distinguishes cells undergoing hyperglycolysis induced by oligomycin A from untreated cells by monitoring their extracellular acidification. To facilitate pH sensing in each droplet, we encapsulate a cell-impermeable pH probe whose fluorescence intensities are quantified. Using this approach, we can differentiate hyperglycolytic cells and concurrently observe single cell heterogeneity in extracellular acidification dynamics. This high-throughput system will be useful in applications that require dynamic phenotyping of single cells with significant heterogeneity.

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