Microfluidic device to study arterial shear-mediated platelet-surface interactions in whole blood: reduced sample volumes and well-characterised protein surfaces.
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.
Kent
Nigel J
NJ
Biomedical Diagnostics Institute, Dublin City University, Glasnevin Dublin 9, Ireland. nigel.kent@dit.ie
Basabe-Desmonts
Lourdes
L
Meade
Gerardene
G
MacCraith
Brian D
BD
Corcoran
Brian G
BG
Kenny
Dermot
D
Ricco
Antonio J
AJ
D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies / Q000276:immunology; D001158:Arteries / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D001696:Biomechanical Phenomena; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D001798:Blood Proteins / Q000276:immunology / Q000378:metabolism*; D001810:Blood Volume; D004867:Equipment Design; D006801:Humans; D007091:Image Processing, Computer-Assisted; D055595:Mechanical Phenomena; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D018625:Microscopy, Atomic Force; D010973:Platelet Adhesiveness; D011485:Protein Binding; D015203:Reproducibility of Results; D012984:Software; D014841:von Willebrand Factor / Q000378:metabolism
D000906:Antibodies; D001798:Blood Proteins; D014841:von Willebrand Factor