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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
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Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated <t>microscope</t> inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
Motion Gui Matlab Code, supplied by MathWorks Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated microscope inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also <xref ref-type=Figure S1 . " width="100%" height="100%">

Journal: Cell Reports

Article Title: High-Content Quantification of Single-Cell Immune Dynamics

doi: 10.1016/j.celrep.2016.03.033

Figure Lengend Snippet: Automated Microfluidic System for Multiparameter Analysis of Single-Cell Immune Dynamics (A) Top left: microfluidic device with functional areas indicated. The scale bar represents 5 mm. Middle right: close-up of a single functional column. The arrows show medium movement during sampling (solid) and circulation for mixing during binding (dashed). Right: close-ups showing a cell captured inside a culture chamber (red), a binding chamber (blue) with a retained bead, and beads inside storage chambers (red). Bottom left: fluorescent imaging of NF-κB activation (top row). The arrow shows the activated cell (RelA in nucleus), and the scale bar represents 20 μm. Middle row: phase contrast images. Bottom row: brightfield-fluorescence merged images of beads. The red spot in the third panel indicates fluorescent sandwich immune assay detection of secreted protein. (B) Setup of microfluidic device mounted on automated microscope inside atmospheric enclosure and image of control software interface. (C) Operation of secretion assay. Beads are initially loaded into storage chambers and then moved by direction of control software to binding chamber. There they are exposed to medium coming from cells through peristaltic pumping. Once analyte is mixed and bound, beads are rinsed and moved back to original storage chambers. This process is repeated with a new bead for different time points so that each row corresponds to a specific time point. See also Figure S1 .

Article Snippet: Imaging and positioning are under control of microscope software (Nikon), while fluidic control of the microfluidic system is directed by way of custom GUI-based MATLAB code ( , ) ( B).

Techniques: Functional Assay, Sampling, Binding Assay, Imaging, Activation Assay, Fluorescence, Microscopy, Control, Software