Pneumococci bind to VWF strings generated in continuous flow. (A) HMW VWF fiber generation was microscopically quantified after exposing confluently grown HUVEC to shear stress using a microfluidic device (ibidi®) at 10 dyne/cm2. FITC-conjugated VWF-specific antibodies detected HMW VWF strings. White arrows point to red fluorescent pneumococci attached to long VWF strings. (B) Attachment of RFP-expressing pneumococci to green fluorescent VWF strings was microscopically observed after 30 min in constant flow (white arrows) and was confirmed by software-based evaluation of fluorescence intensities of the defined ROI. Pictures were taken in real time using the fluorescence equipment of a confocal laser scanning microscope (SP8, Leica). Scale bar represents 10 μm.
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