Multiplexed profiling of single-cell secretion on a hierarchical loading microwell chip

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Protein secretion is crucial for cellular communication, development, and immune responses.
• Accurate detection of single-cell protein secretion is essential but technically challenging.
• Multiplexed analysis of secreted proteins at the single-cell level is highly desired.

Purpose of the Study:

• To develop a novel method for highly multiplexed detection of single-cell protein secretion.
• To demonstrate the platform's utility in profiling cytokine secretion from immune cells.
• To enable advanced single-cell analysis for biological and clinical applications.

Main Methods:

• A hierarchical loading microwell chip (HL-Chip) was designed for efficient single-cell and microbead alignment.
• The platform facilitates the capture of secreted proteins from thousands of single cells.
• Multiplexed detection of six cytokines from single T cells and macrophages was performed.

Main Results:

• The HL-Chip method successfully enabled highly multiplexed detection of secreted proteins.
• Profiling of six cytokines revealed transient cytokine bursts and polyfunctional heterogeneity in stimulated T cells.
• Antigen-specific stimulation of T cell receptor-engineered T (TCR-T) cells showed distinct secretion patterns.

Conclusions:

• The HL-Chip offers a simple and versatile platform for highly multiplexed single-cell secretion analysis.
• This technology advances the study of cellular communication and immune responses.
• Potential applications include biological discovery and clinical diagnostics for various diseases.