Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture
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Ploegh_Longitudinal.pdf
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Author(s)
Servos, Mariah M.
Dougan, Stephanie K.
Dura, Burak
Barry, Rachel M
Ploegh, Hidde
Voldman, Joel
Date Issued
June 2016
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
Dura, Burak et al. “Longitudinal Multiparameter Assay of Lymphocyte Interactions from Onset by Microfluidic Cell Pairing and Culture.” Proceedings of the National Academy of Sciences 113.26 (2016): E3599–E3608. © 2016 National Academy of Sciences
Version
Final published version
Abstract
Resolving how the early signaling events initiated by cell–cell interactions are transduced into diverse functional outcomes necessitates correlated measurements at various stages. Typical approaches that rely on bulk cocultures and population-wide correlations, however, only reveal these relationships broadly at the population level, not within each individual cell. Here, we present a microfluidics-based cell–cell interaction assay that enables longitudinal investigation of lymphocyte interactions at the single-cell level through microfluidic cell pairing, on-chip culture, and multiparameter assays, and allows recovery of desired cell pairs by micromanipulation for off-chip culture and analyses. Well-defined initiation of interactions enables probing cellular responses from the very onset, permitting single-cell correlation analyses between early signaling dynamics and later-stage functional outcomes within same cells. We demonstrate the utility of this microfluidic assay with natural killer cells interacting with tumor cells, and our findings suggest a possible role for the strength of early calcium signaling in selective coordination of subsequent cytotoxicity and IFN-gamma production. Collectively, our experiments demonstrate that this new approach is well-suited for resolving the relationships between complex immune responses within each individual cell.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Microsystems Technology Laboratories
Massachusetts Institute of Technology. Research Laboratory of Electronics
Whitehead Institute for Biomedical Research
Koch Institute for Integrative Cancer Research at MIT
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1073/pnas.1515364113
