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. 2019 Nov 4;9:15932. doi: 10.1038/s41598-019-52041-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Rheological effects of adherent neutrophils on platelet aggregation under flow. Interactions of fluorescence-labeled platelets were analyzed in vitro in custom-made PDMS microfluidic devices as detailed in Methods, a schematic illustration of the device (A) and transillumination microscopy images of the flow channels (B) are shown (arrows indicate individual bump structures). Panel (C) shows quantitative data for the number of platelets adhering to the wall of flow channels exhibiting varying densities of bump structures (mimicking intravascularly adherent neutrophils) after 10 min of perfusion with mouse blood. In selected experiments, anti-GPIbα mAbs were added to the blood (mean ± SEM for n = 3–4 per group; *p < 0.05 vs. flat devices, ^§p < 0.05 vs. lowest bump density, ^#p < 0.05 vs. highest bump density). Interactions of fluorescence-labeled platelets with bump structures were analyzed in vitro in custom-made PDMS microfluidic devices by high-resolution microscopy as detailed in Methods, representative images are shown (D; scale bar: 20 μm). In panel (E; scale bar: 10 μm), formation of GPIbβ-positive tethers in platelets (red) adhering to bump structures is shown.