A high-throughput modular microfluidic platform for versatile functional assays at single-cell level

Area of Science:

• Single-cell biology
• Microfluidics
• Functional genomics

Background:

• Current single-cell technologies offer limited functional insights, providing only static molecular data.
• Understanding dynamic cellular behavior is crucial for deciphering biological complexity and disease mechanisms.

Purpose of the Study:

• To introduce the High-throughput Single-Cell Omni-functional Profiling Engine (HiSCOPE) for dynamic single-cell functional profiling.
• To demonstrate HiSCOPE's capability in analyzing diverse cellular functions and interactions.

Main Methods:

• Development of a microfluidic platform (HiSCOPE) with hydrodynamic trapping for single cells, pairs, and trios.
• Integration of modular assay chambers to support various functional assays (viability, proliferation, cytotoxicity, cytokine secretion, cell-cell interactions).
• Incorporation of live-cell retrieval for downstream molecular analysis.

Main Results:

• HiSCOPE successfully profiled functional dynamics of over 10,000 single cells.
• Demonstrated utility in evaluating drug susceptibility in leukemia cells and NK cell cytotoxicity.
• Dissected T cell-macrophage crosstalk, revealing functional diversity masked in bulk assays.

Conclusions:

• HiSCOPE provides a scalable solution for high-throughput functional profiling of single cells.
• The platform enables deeper understanding of cellular heterogeneity and interactions.
• HiSCOPE has broad implications for advancing immunotherapy and personalized medicine.