Probing cellular heterogeneity in cytokine-secreting immune cells using droplet-based microfluidics

Abstract

Here, we present a platform to detect cytokine (IL-2, IFN-γ, TNF-α) secretion of single, activated T-cells in droplets over time. We use a novel droplet-based microfluidic approach to encapsulate cells in monodisperse agarose droplets together with functionalized cytokine-capture beads for subsequent binding and detection of secreted cytokines from single cells. This method allows high-throughput detection of cellular heterogeneity and maps subsets within cell populations with specific functions.

Malondialdehyde, Adiponectin, Nitric Oxide, C-Reactive Protein, Tumor Necrosis Factor-Alpha and Insulin Resistance Relationships and Inter-Relationships in Type 2 Diabetes Early Stage Is Metformin Alone Adequate in This Stage

“…Moreover, as previously reported by [28] high MDA can induce many inflammatory cytokines. These cytokines trigger others production and lead to more oxidative stress [29]. Upon the aforementioned, in a way or another diabetes induces lipid peroxidation, lipid peroxidation stimulates inflammation and inflammation, in turn, stimulates more lipid peroxidation supporting the involvement of oxidative stress in chronic inflammation.…”

Applications of Microfluidics in Quantitative Biology

“…This tunability allows the high throughput feature to be applied in many fields, including biochemical assays, [72] cell-based enzymatic assays, [73] DNA amplification, [74] cell sorting, [75,76] microspheres, [77] nanoparticles, [78] core-shell microcapsule synthesis [79] so on. As a detailed example, cellular heterogeneity arising from the stochasticity of gene expression, protein function and metabolite quantity is crucial for cell behavior and evolution.…”

“…This system encapsulates cells together with functionalized cytokine-capture beads for subsequent binding, and for following detection of secreted cytokines at single-cell level. [77] The Mazutis group used droplet-based microfluidics to capture thousands of individual cells with unbiased diversity and performed the whole transcriptome and genomic analysis successfully. [83] The Weitz group developed an ultrahigh-throughput screening platform for directed evolution to discover variants of the target enzyme.…”

Continuous measurement of enzymatic kinetics in droplet flow for point-of-care monitoring

“…The carrier fluid used was FC-40 fluorinated oil (3 M, UK) containing a 0.35% w/w concentration of non-ionic tri-block copolymer surfactant synthesised in-house. 29 Syringe pumps (PHD 2000/Harvard Apparatus) were used to pump all fluids, with the reagent mix and glucose solutions injected at the same flow rate and the flow rate of oil (1.31 µL min −1 ) tuned to achieve droplets measuring approximately 10 nL. Following generation, the droplets flowed into a polytetrafluoroethylene (PTFE) tubing (0.4 mm ID, 0.7 mm OD, Adtech Polymer Engineering Ltd) connected to the flow cell for optical detections.…”