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Abstract Integrated microfluidic cellular phenotyping platforms provide a promising means of studying a variety of inflammatory diseases mediated by cell‐secreted cytokines. However, immunosensors integrated in previous microfluidic platforms lack the sensitivity to detect small signals in the cellular secretion of proinflammatory cytokines with high precision. This limitation prohibits researchers from studying cells secreting cytokines at low abundance or existing at a small population. Herein, the authors present an integrated platform named the “digital Phenoplate (dPP),” which integrates digital immunosensors into a microfluidic chip with on‐chip cell assay chambers, and demonstrates ultrasensitive cellular cytokine secretory profile measurement. The integrated sensors yield a limit of detection as small as 0.25 pg mL−1for mouse tumor necrosis factor alpha (TNF‐α). Each on‐chip cell assay chamber confines cells whose population ranges from ≈20 to 600 in arrayed single‐cell trapping microwells. Together, these microfluidic features of the dPP simultaneously permit precise counting and image‐based cytometry of individual cells while performing parallel measurements of TNF‐α released from rare cells under multiple stimulant conditions for multiple samples. The dPP platform is broadly applicable to the characterization of cellular phenotypes demanding high precision and high throughput.
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Airplug‐Mediated Isolation and Centralization of Single T Cells in Rectangular Microwells for Biosensing
Abstract Sorting cells in a single cell per microwell format is of great interest to basic biological 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 therapeutic applications. The present study, 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 toward the center of the rectangular microwells is reported. The chip includes 2250 microwells (each 500 × 50 × 20 µm3) arranged in nine rows. Results show 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.
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- Award ID(s):
- 1701322
- PAR ID:
- 10456111
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Advanced Therapeutics
- Volume:
- 3
- Issue:
- 1
- ISSN:
- 2366-3987
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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