Related Experiment Video
Updated: Jul 8, 2025
09:43
Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses
Published on: March 8, 2024
1.7K
Single-Cell Secretion Analysis via Microfluidic Cell Membrane Immunosorbent Assay for Immune Profiling.
Ying Xu1,2, Ka-Chun Wu3,4, Wenxin Jiang1,2
1Department of Biomedical Engineering, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon Tong, Hong Kong SAR 999077, China.
Analytical Chemistry
|December 18, 2023
Summary
A new assay enables high-throughput single-cell analysis of immune cell secretions and surface proteins. This method advances precision medicine by identifying distinct immune cell profiles in healthy donors and nasopharyngeal carcinoma patients.
Area of Science:
- Biomedical Engineering
- Immunology
- Analytical Chemistry
Background:
- Single-cell phenotypic analysis is crucial for precision medicine, but screening cell secretions remains difficult.
- Existing methods struggle with high-throughput, multiplexed analysis of individual cell functions.
- Cell secretions are key indicators of immune cell activity and function.
Purpose of the Study:
- To develop a novel, high-throughput single-cell secretion assay for multiplexed phenotypic analysis.
- To integrate immunosorbent assays and droplet microfluidics for enhanced single-cell screening.
- To validate the assay's capability in analyzing immune cell differentiation and activation.
Main Methods:
- Development of a cell membrane-based assay using cholesterol-linked antibodies (CLAbs).
- Encapsulation of CLAb-grafted single cells in droplets for secretion capture.
- Extraction, fluorescence labeling, and multiplexed measurement of secretions and surface proteins via flow cytometry.
Main Results:
- The assay successfully analyzed THP-1 cells, M1 macrophages, and dendritic cells, indicating differentiation efficiency.
- Peripheral blood mononuclear cells (PBMCs) from healthy donors showed varied active immune cell populations (6.62-47.14%) under stimuli.
- PBMCs from nasopharyngeal carcinoma patients exhibited a higher percentage of actively cytokine-secreting cells in basal state (2.82 ± 1.48%) compared to healthy donors (0.70 ± 0.29%).
Conclusions:
- The developed assay provides a flexible and high-throughput platform for single-cell secretion analysis.
- This method enables precise phenotyping and identification of distinct immune cell populations.
- The findings highlight potential diagnostic biomarkers in nasopharyngeal carcinoma patients based on single-cell immune activity.
