2015
DOI: 10.1073/pnas.1416756112
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Highly multiplexed profiling of single-cell effector functions reveals deep functional heterogeneity in response to pathogenic ligands

Abstract: Despite recent advances in single-cell genomic, transcriptional, and mass-cytometric profiling, it remains a challenge to collect highly multiplexed measurements of secreted proteins from single cells for comprehensive analysis of functional states. Herein, we combine spatial and spectral encoding with polydimethylsiloxane (PDMS) microchambers for codetection of 42 immune effector proteins secreted from single cells, representing the highest multiplexing recorded to date for a single-cell secretion assay. Usin… Show more

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Cited by 260 publications
(284 citation statements)
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“…Interestingly, the overall structure has not been shifted much after LPS stimulation. The MIF subpopulation is completely separated from others, and the heterogeneity is not changed significantly after stimulation 6. This data indicate that MIF secretion is conservative function to macrophages.…”
Section: Resultsmentioning
confidence: 66%
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“…Interestingly, the overall structure has not been shifted much after LPS stimulation. The MIF subpopulation is completely separated from others, and the heterogeneity is not changed significantly after stimulation 6. This data indicate that MIF secretion is conservative function to macrophages.…”
Section: Resultsmentioning
confidence: 66%
“…For example, by simply designing more barcode ssDNA probes and cDNA‐antibody conjugates, the same architecture of the current technology can measure up to 40 different cytokines. Noticeably, the other barcode chips in our group and those in Fan's group have microchamber volumes at least eight times larger for single‐cell cytokine assays 6. Therefore, without losing much sensitivity, this strategy by expansion of microarray size and microwell volume can further increase multiplexity to hundreds.…”
Section: Discussionmentioning
confidence: 91%
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“…For example, NK cells were shown to eliminate local target cells independent of the neighboring NK cells [14], and experiments with multiple target cells identified a particularly active serial killer subpopulation with faster kill dynamics [15]. Similar co-culture studies in combination with barcoded antibody arrays [41,42 ] also enabled quantifying the influence of paracrine signaling on tumor cell functional states and signaling networks with multiplexed detection of intracellular and secreted proteins [18,20,21].…”
Section: Profiling Intercellular Interactionsmentioning
confidence: 99%