Sarıyer Rüya Meltem, Gill Kirandeep, Needs Sarah H, Hodge Daniel, Reis Nuno M, Jones Chris I, Edwards Alexander D
Reading School of Pharmacy, University of Reading Whiteknights Reading RG6 6UB UK
Department of Chemical Engineering and Centre for Biosensors, Bioelectronics and Biodevices (CBio), University of Bath Bath BA2 7AY UK
Sens Diagn. 2023 Oct 17;2(6):1623-1637. doi: 10.1039/d3sd00162h. eCollection 2023 Nov 9.
Measuring the complex processes of blood coagulation, haemostasis and thrombosis that are central to cardiovascular health and disease typically requires a choice between high-resolution low-throughput laboratory assays, or simpler less quantitative tests. We propose combining mass-produced microfluidic devices with open-source robotic instrumentation to enable rapid development of affordable and portable, yet high-throughput and performance haematological testing. A time- and distance-resolved fluid flow analysis by Raspberry Pi imaging integrated with controlled sample addition and illumination, enabled simultaneous tracking of capillary rise in 120 individual capillaries (∼160, 200 or 270 μm internal diameter), in 12 parallel disposable devices. We found time-resolved tracking of capillary rise in each individual microcapillary provides quantitative information about fluid properties and most importantly enables quantitation of dynamic changes in these properties following stimulation. Fluid properties were derived from flow kinetics using a pressure balance model validated with glycerol-water mixtures and blood components. Time-resolved imaging revealed fluid properties that were harder to determine from a single endpoint image or equilibrium analysis alone. Surprisingly, instantaneous superficial fluid velocity during capillary rise was found to be largely independent of capillary diameter at initial time points. We tested if blood function could be measured dynamically by stimulating blood with thrombin to trigger activation of global haemostasis. Thrombin stimulation slowed vertical fluid velocity consistent with a dynamic increase in viscosity. The dynamics were concentration-dependent, with highest doses reducing flow velocity faster (within 10 s) than lower doses (10-30 s). This open-source imaging instrumentation expands the capability of affordable microfluidic devices for haematological testing, towards high-throughput multi-parameter blood analysis needed to understand and improve cardiovascular health.
测量对心血管健康和疾病至关重要的血液凝固、止血和血栓形成等复杂过程，通常需要在高分辨率低通量实验室检测或更简单的非定量检测之间做出选择。我们建议将大规模生产的微流控设备与开源机器人仪器相结合，以实现经济实惠且便携、同时高通量且高性能的血液学检测的快速开发。通过集成控制样品添加和照明的树莓派成像进行时间和距离分辨的流体流动分析，能够在12个平行的一次性设备中同时跟踪120根内径约为160、200或270μm的单个毛细管中的毛细管上升情况。我们发现对每个微毛细管中毛细管上升的时间分辨跟踪提供了有关流体特性的定量信息，最重要的是能够对刺激后这些特性的动态变化进行定量。流体特性是使用通过甘油 - 水混合物和血液成分验证的压力平衡模型从流动动力学中推导出来的。时间分辨成像揭示了仅从单个终点图像或平衡分析中难以确定的流体特性。令人惊讶的是，在初始时间点，毛细管上升过程中的瞬时表面流体速度在很大程度上与毛细管直径无关。我们测试了是否可以通过用凝血酶刺激血液来触发整体止血激活，从而动态测量血液功能。凝血酶刺激使垂直流体速度减慢，这与粘度的动态增加一致。这种动态变化是浓度依赖性的，最高剂量比低剂量（10 - 30秒）更快（在10秒内）降低流速。这种开源成像仪器扩展了经济实惠的微流控设备用于血液学检测的能力，朝着理解和改善心血管健康所需的高通量多参数血液分析发展。
