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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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Abstract

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.

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Notes

  1. Some protein is expected to adsorb onto the PMMA surface; however, this should not affect the results, as the depth of focus, which is centered on the fluid-contacting surface of the glass bottom plate, is only 20 μm (objective working distance = 190 μm, slide thickness = 170 μm); furthermore, the flow conditions are such that any platelet that interacts with a side wall or the top plate will not reach the bottom plate within the microscope’s field of view.

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Acknowledgements

The authors gratefully acknowledge the help of Dr. Cedric Volcke with the AFM measurements, and related discussions, and the help of Dr. Helen Berney and Dr. Arun Harish, Field Applications Scientist, Farfield Group Ltd., UK with the dual-polarized interferometry and related discussions. This material is based upon works supported by the Science Foundation Ireland under Grant No. 05/CE3/B754.

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Authors and Affiliations

  1. Biomedical Diagnostics Institute, Dublin City University, Glasnevin, Dublin 9, Ireland

    Nigel J. Kent, Lourdes Basabe-Desmonts, Brian D. MacCraith & Antonio J. Ricco

  2. Biomedical Diagnostics Institute, Dept of Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, 123 St Stephens Green, Dublin 2, Ireland

    Gerardene Meade & Dermot Kenny

  3. School of Mechanical and Manufacturing Engineering, Dublin City University, Glasnevin, Dublin 9, Ireland

    Brian G. Corcoran

  4. The Biomedical Devices and Assistive Technology Research Group, College of Engineering and Built Environment, Dublin Institute of Technology, Bolton St., Dublin 1, Ireland

    Nigel J. Kent

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  1. Nigel J. Kent
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  2. Lourdes Basabe-Desmonts
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  3. Gerardene Meade
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  6. Dermot Kenny
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Correspondence to Nigel J. Kent or Antonio J. Ricco.

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Fig. S1

3D height AFM images of 2 × 2 μm2 areas of VWF protein layers on SiO2-on-Si wafers for 2-hr adsorption from aqueous VWF solutions: 5, 50, 100, and 500 μg/mL. These correspond to the four 2D images shown in Figure 6(c). (JPEG 259 kb)

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Kent, N.J., Basabe-Desmonts, L., Meade, G. et al. Microfluidic device to study arterial shear-mediated platelet-surface interactions in whole blood: reduced sample volumes and well-characterised protein surfaces. Biomed Microdevices 12, 987–1000 (2010). https://doi.org/10.1007/s10544-010-9453-y

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