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

Area of Science:

• Biotechnology
• Cell Biology
• Analytical Chemistry

Background:

• Quantitative high-throughput metabolism assays are crucial for single-cell phenotyping dynamics.
• Extracellular acidification is a key metabolic phenotype linked to cellular functions like tumorigenicity.

Purpose of the Study:

• To develop a versatile microfluidic system for single-cell optical pH analysis (SCO-pH).
• To enable high-throughput, temporal extracellular pH analysis for dynamic single-cell phenotyping.

Main Methods:

• Compartmentalizing single cells in 140-pL droplets within a microfluidic system.
• Immobilizing approximately 40,000 droplets in a 2D array for temporal analysis.
• Encapsulating a cell-impermeable pH probe for fluorescence-based pH quantification within each droplet.

Main Results:

• SCO-pH successfully distinguishes cells with induced hyperglycolysis from untreated cells by monitoring extracellular acidification.
• The system enables concurrent observation of single-cell heterogeneity in extracellular acidification dynamics.
• Demonstrated differentiation of hyperglycolytic cells using temporal pH analysis.

Conclusions:

• The developed SCO-pH system provides a high-throughput method for dynamic single-cell metabolic phenotyping.
• This technology is valuable for applications requiring the analysis of single-cell heterogeneity.
• SCO-pH advances the study of cellular metabolism and its implications in various biological contexts.