Microfluidic Core Facility

微流控核心设施

• 批准号: 8375805
• 负责人: Alex Groisman
• 金额: $ 11.76万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8375805
• 关键词: Adhesions; Area; Biological Assay; Blood; Blood Platelets; Blood Vessels; Blood flow; Blood specimen; Cell Adhesion; Cell Adhesion Molecules; Collagen; Core Facility; Devices; Endothelium; Extracellular Matrix; Feedback; Fibrinogen; Human Resources; Injury; Integrins; Laboratories; Laboratory Personnel; Leukocytes; Maintenance; Mediating; Microfabrication; Microfluidic Microchips; Microfluidics; Microscopy; P-Selectin; Pattern; Performance; Perfusion; Platelet aggregation; Play; Production; Research Personnel; Role; Site; Spatial Distribution; Surface; System; Techniques; Testing; Thrombosis; Training; Vascular Cell Adhesion Molecule-1; Vascular Diseases; Work; base; constriction; density; design; improved; in vivo; neutrophil; programs; research study; shear stress; von Willebrand Factor

Core B (the Microfluidics Core) will design and build microfluidic devices and flow control systems, develop advanced substrate-coating techniques, and work with the laboratories participating in the Program to apply them to studies of surface adhesion of platelet and leukocytes. Core B will set up, upgrade, conjugate with microscopy, and maintain microfluidic systems in the laboratories of the Program, will train the personnel of the laboratories and take part in pilot experiments with the systems. Core B will fabricate microfluidic devices, supply them to the Program laboratories, and modify the devices and techniques based on the feedback from the laboratories. For experiments on substrate adhesion and aggregation of platelets (Projects 1 - 3), Core B will build microfluidic devices subjecting blood to two to three orders-of-magnitude ranges of shear stress in each assay, with shear stress reaching >1000 dynes/cm^2 for experiments with von Willebrand factor, VWF, (Project 3) and with small amounts of blood (<100 uL for Projects 1 and 2 and <400 uL for Project 3) required for an assay. Platelet adhesion from two different blood samples to identical substrates will be tested in parallel. Substrate coating techniques will be developed to control the site densities of adhesion molecules (collagen, fibrinogen, and VWF) and to test the adhesion to substrates with a range of site densities in a single assay. Micro-channels with constrictions will be fabricated to study the effect of extensional vs. shear flow on the activity of VWF and on VWF-mediated adhesion and aggregation of platelets (Project 3). For experiments on rolling and arrest of leukocytes (Projects 1), Core B will build microfluidic devices with a range of shear stresses and customized substrate coatings. Mixed substrate coatings will be created with site densities of different adhesion molecules (e.g., P-selectin and VCAM-1) independently varied and controlled. Substrates with sharp boundaries between differently coated areas will be produced to test for possible integrin activation by adhesion molecules and to study the dynamics of neutrophil arrest (Project 1). Micro-patterned substrates with ~1um-wide differently coated strips will be made to emulate non-uniform distributions of adhesion molecules expressed on endothelium.

Core B（微流体核心）将设计和建立微流体设备和流控制系统，开发先进的底物涂层技术，并与参与程序的实验室合作，将其应用于血小板和白细胞表面粘附研究。 Core B将在程序实验室中建立，升级，与显微镜结合并维护微流体系统，将培训实验室人员的人员，并参与系统的试点实验。核心B将制造微流体设备，将其提供给程序实验室，并根据实验室的反馈来修改设备和技术。对于底物粘附和血小板聚集的实验（项目1-3），核心B将在每个测定中构建血液受到两到三个剪切应力的血液的微流体范围，剪切应力范围> 1000 dynes/cm^2用于实验> 1000 dynes/cm^forle forle wille willbrand corvation 1 and forly，vwf，vwf and（vw forly）（vw forly）（<vwf），<vwf forly（vwf fllbrand consect and）（<vwffff florl），<vwf florl（<）（<） <400 UL对于项目3）进行测定。从两个不同的血液样本到相同底物的血小板粘附将平行测试。将开发底物涂料技术来控制粘附分子（胶原蛋白，纤维蛋白原和VWF）的位点密度，并在单个测定中测试具有一系列位点密度的底物的粘附。将制造具有收缩的微通道，以研究延伸性与剪切流对VWF活动以及VWF介导的粘附和血小板聚集的影响（项目3）。对于白细胞滚动和逮捕的实验（项目1），Core B将建造具有一系列剪切应力和定制底物涂层的微流体设备。混合的底物涂层将与不同粘附分子（例如P-选择蛋白和VCAM-1）的位点密度独立变化和控制。将产生不同涂层区域之间具有尖锐边界的底物，以测试通过粘附分子的可能整合素激活，并研究中性粒细胞停滞的动力学（项目1）。将制作具有〜1um宽的涂层条带的微图案底物，以模拟内皮上表达的粘附分子的不均匀分布。