Fluxion Biosciences, 384 Oyster Point Blvd., Suite 6, South San Francisco, California 94080, USA.
Biotechnol Bioeng. 2011 Dec;108(12):2978-87. doi: 10.1002/bit.23243. Epub 2011 Jul 16.
The study of platelet behavior in real-time under controlled shear stress offers insights into the underlying mechanisms of many vascular diseases and enables evaluation of platelet-focused therapeutics. The two most common methods used to study platelet behavior at the vessel wall under uniform shear flow are parallel plate flow chambers and cone-plate viscometers. Typically, these methods are difficult to use, lack experimental flexibility, provide low data content, are low in throughput, and require large reagent volumes. Here, we report a well plate microfluidic (WPM)-based system that offers high throughput, low reagent consumption, and high experimental flexibility in an easy to use well plate format. The system consists of well plates with an integrated array of microfluidic channels, a pneumatic interface, an automated microscope, and software. This WPM system was used to investigate dynamic platelet behavior under shear stress. Multiple channel designs are presented and tested for shear loads with whole blood to determine their applicability to study thrombus formation. Normal physiological shear (0.1-20 dyn/cm(2) ) and pathological shear (20-200 dyn/cm(2) ) devices were used to study platelet behavior in vitro under various shear, matrix coating, and monolayer conditions. The high physiological relevance, low blood consumption, and increased throughput create a valuable technique available to vascular biology researchers. The approach also has extensibility to other research areas including inflammation, cancer biology, and developmental/stem cell research.
实时研究受控切应力下的血小板行为可深入了解许多血管疾病的潜在机制，并能评估针对血小板的治疗方法。在均一剪切流条件下研究血管壁处血小板行为的两种最常用的方法是平行板流室和锥板黏度计。通常，这些方法难以使用，缺乏实验灵活性，提供的数据内容低，通量低，且需要大量试剂。在此，我们报告了一种基于微孔板的微流控（WPM）系统，该系统在易于使用的微孔板格式中提供高通量、低试剂消耗和高实验灵活性。该系统由带有集成微流道阵列的微孔板、气动接口、自动化显微镜和软件组成。该 WPM 系统用于研究剪切应力下的动态血小板行为。提出并测试了多个通道设计，用于全血的剪切负载，以确定其在血栓形成研究中的适用性。使用正常生理剪切（0.1-20 dyn/cm(2)）和病理剪切（20-200 dyn/cm(2)）装置，在各种剪切、基质涂层和单层条件下研究体外血小板行为。这种高度的生理相关性、低血液消耗和增加的通量为血管生物学研究人员创造了一种有价值的技术。该方法还可扩展到其他研究领域，包括炎症、癌症生物学和发育/干细胞研究。
