Biomedical Diagnostics Institute, Dublin City University, Glasnevin Dublin 9, Ireland.
Biomed Microdevices. 2010 Dec;12(6):987-1000. doi: 10.1007/s10544-010-9453-y.
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 μL of whole human blood per assay. The device's parallel-plate flow chamber, with 0.1 mm² 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 cm² 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.
我们报告了一种新的设备,可以在控制的流体剪切条件下分析蛋白质表面上的细胞表面相互作用。通过使用每分析物仅 200μL 的全人血在动脉血管剪切速率下研究与固定化 von Willebrand 因子相互作用的血小板,证明了其性能。该设备的平行板流室具有 0.1mm² 的横截面积和 1:40 的高宽比,在芯片表面上提供均匀、明确定义的剪切率,同时对流体流动形态几乎没有垂直壁效应,从而最小化样品体积流量。通过椭圆测量法、原子力显微镜和荧光免疫染色证明了涂层工艺,可在 0.7cm² 的细胞表面相互作用区域上提供可重复、均匀、一致的蛋白质层。定制的图像处理可以量化整个全血流动分析过程中动态细胞表面覆盖率与时间的关系,以用于给定的药物治疗或疾病状态。该设备可以跟踪抗血小板药物的剂量反应,适用于即时诊断,并设计用于适应大规模生产。