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Updated: Dec 3, 2025
A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
Published on: June 16, 2016
Airplug-mediated isolation and centralization of single T cells in rectangular microwells for biosensing
Pavithra Sukumar1, Muhammedin Deliorman1, Ayoola T Brimmo1
1Division of Engineering, New York University Abu Dhabi (NYUAD), P.O. Box 129188, Abu Dhabi, UAE.
Insights
This study introduces a novel method using airplugs to center single cells in microwells, improving cytokine detection for T cell research and biotherapeutics.
Area of Science:
- Biotechnology
- Cell Biology
- Biosensing
Background:
- Single-cell analysis is crucial for basic biology, biotherapeutics, and biosensing.
- Isolating individual immune T cells enables multiplex cytokine profiling for therapeutic applications.
- Random cell positioning in microwells introduces bias in temporal cytokine sensing.
Purpose of the Study:
- To eliminate bias in temporal cytokine sensing by precisely localizing single cells.
- To develop a method for aligning cells within microwells relative to sensing elements.
- To enable unbiased, real-time multiplex cytokine detection from single T cells.
Main Methods:
- Utilized in situ formation and release of airplugs for cell localization.
- Employed a chip with 2250 rectangular microwells (500 × 50 × 20 μm³).
- Arranged microwells in 9 rows for high-throughput analysis.
Main Results:
- Achieved 20% efficiency in trapping single T cells per microwell.
- Demonstrated precise cell localization within ±3% of the microwell center.
- Successfully mitigated positional bias in temporal cytokine sensing.
Conclusions:
- The developed airplug-based platform enables accurate and unbiased single-cell cytokine detection.
- This technology supports advanced phenotyping and biotherapeutics studies.
- Provides a foundation for real-time dynamic analysis of single immune cells.
Abstract:
Sorting cells in a single cell per microwell format is of great interest to basic biology studies, biotherapeutics, and biosensing including cell phenotyping. For instance, isolation of individual immune T cells in rectangular microwells has been shown to empower the multiplex cytokine profiling at the single cell level for therapeutics applications. The present study, however, shows that there is an existing bias in temporal cytokine sensing that originates from random "unpredicted" positions of loaded cells within the rectangular microwells. To eliminate this bias, the isolated cells need to be well-aligned with each other and relative to the sensing elements. Hence, an approach that utilizes the in situ formation and release of airplugs to localize cells towards the center of the rectangular microwells is reported. The chip includes 2250 microwells (each 500 × 50 × 20 μm3) arranged in 9 rows. Results showed 20% efficiency in trapping single T cells per microwells, where cells are localized within ±3% of the center of microwells. The developed platform could provide real-time dynamic and unbiased multiplex cytokine detection from single T cells for phenotyping and biotherapeutics studies.
