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The gating strategy for all samples was to set a gate on the unstimulated control such that CD3+CD8+CD4-T cells were negative for IFN-γ and TNF-α.
In analyzing cells positively transfected for tagYFP, we set a gate based on untransfected cells (Supporting Information Figure 4).
To calculate the iMFI we set a gate that collects events over the background fluorescence threshold (Supporting Information Figure 4).
Monoclonal antibodies against CD-14 antigen (anti-CD-14-PE, Immuno Quality Products, Groningen, the Netherlands) were used to set a gate for monocytes.
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Data for 10 cells were collected for each sample and prior to data collection cell debris was excluded by setting a gate on a forward vs side scatter two-dimensional dot plot.
Then you set an age gate, and are able to specify what kind of data you want to collect.
To detect the Treg population, the first gate was set for mononuclear cells based on the profiles of their cell size, and then set a quadro gate to separate negative and positive populations for CD4 and CD25.
That alone would be pretty impressive in a device that's priced at a $29 pledge to start for backers, but it also offers a Bluetooth proximity alarm sensor, which makes it possible to set a location gate so that you can receive alerts if you move away from your camera, or whatever else it's attached to.
The platelets were gated by setting a polygonal gate and transferred to a FL-1 (CD62P-FITC) and FL-2 (CD41-PE) histogram.
Dependent on their specific FSC and SSC characteristics monocytes and granulocytes were gated by setting a polygonal gate and were individually displayed in FL-2 versus FSC dot plots for analysis of platelet-monocyte and platelet-granulocyte complexes.
INSIDE: The house -- set behind a gate flanked by stone lions -- was built in 1896, and restored in 1998 by its current owners.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com