Microfluidic device to study arterial shear-mediated platelet-surface interactions in whole blood: reduced sample volumes and well-characterised protein surfaces
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作者:
Kent, Nigel J.
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Dublin City Univ, Biomed Diagnost Inst, Glasnevin Dublin 9, Ireland
Dublin Inst Technol, Biomed Devices & Assist Technol Res Grp, Coll Engn & Built Environm, Dublin 1, IrelandDublin City Univ, Biomed Diagnost Inst, Glasnevin Dublin 9, Ireland
Kent, Nigel J.
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Basabe-Desmonts, Lourdes
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Dublin City Univ, Biomed Diagnost Inst, Glasnevin Dublin 9, IrelandDublin City Univ, Biomed Diagnost Inst, Glasnevin Dublin 9, Ireland
We report a novel device to analyze cell-surface interactions under controlled fluid-shear conditions on well-characterised protein surfaces. Its performance is demonstrated by studying platelets interacting with immobilised von Willebrand Factor at arterial vascular shear rates using just 200 mu L of whole human blood per assay. The device's parallel-plate flow chamber, with 0.1 mm(2) cross sectional area and height-to-width ratio of 1:40, provides uniform, well-defined shear rates along the chip surface with negligible vertical wall effects on the fluid flow profile while minimizing sample volumetric flow. A coating process was demonstrated by ellipsometry, atomic force microscopy, and fluorescent immunostaining to provide reproducible, homogeneous, uniform protein layers over the 0.7 cm(2) cell-surface interaction area. Customized image processing quantifies dynamic cellular surface coverage vs. time throughout the whole-blood-flow assay for a given drug treatment or disease state. This device can track the dose response of anti-platelet drugs, is suitable for point-of-care diagnostics, and is designed for adaptation to mass manufacture.