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Interfacing droplet microfluidics with antibody barcodes for multiplexed single-cell protein secretion profiling
Tahereh Khajvand1, Peifeng Huang1, Linmei Li2
1The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, State Key Laboratory of Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China. zhuzhi@xmu.edu.cn.
This study introduces a novel droplet microfluidic device for analyzing protein secretions from individual cells. The platform enhances single-cell capture efficiency and enables multiplexed analysis, offering a powerful tool for understanding cellular heterogeneity in health and disease.
Area of Science:
- Biotechnology
- Microfluidics
- Cellular Biology
Background:
- Single-cell protein secretion analysis is crucial for understanding cellular heterogeneity in health and disease.
- Existing platforms face limitations like low cell occupancy, reagent waste, and sensitivity issues.
Purpose of the Study:
- To develop an integrated droplet microfluidic device for multiplexed single-cell secretome analysis.
- To overcome limitations of current single-cell analysis platforms.
Main Methods:
- Utilized a droplet microfluidic device with spatially patterned antibody barcodes.
- Employed a trapping array of 100 picoliter-sized isolation chambers for high single-cell capture efficiency (>80%) and viability (>90%).
- Validated the platform by detecting four multiplexed cytokines (IL-8, MCP-1, MIP-1b, TNF-a/IL-10) from human macrophages.
Main Results:
- Achieved high single-cell capture efficiency and viability in the microfluidic device.
- Successfully performed multiplexed cytokine detection in stimulated and unstimulated human macrophages.
- Demonstrated the platform's utility in profiling protein secretions from cancer cell lines and patient-derived cancer cells, revealing secretion heterogeneity.
Conclusions:
- The developed microfluidic platform enables efficient, multiplexed single-cell secretome analysis.
- Offers a reliable and streamlined workflow for protein secretion assays with low cell numbers and short incubation times (~4 h).
- Has broad applications for studying secretion-mediated cellular heterogeneity in various biological contexts.
