Low-Cost Digital Imaging System for Evaluating Thrombosis

描述(申请人提供)：临床血栓形成对数百万患者的健康有一系列和广泛的影响，如心脏病发作、中风、静脉血栓栓塞症和弥漫性血管内凝血。血栓形成的实验研究需要考虑单个物理场所内的许多汇聚因素--血管内血栓。凝血因子和血小板激活/积聚的相互作用，以及每种因素在动脉或静脉或微血管血栓形成中的相对参与--不同的实体具有不同的血栓形成和溶解机制--尚不完全清楚。为了克服最近开发的血栓研究系统的高成本和劳动密集型方面的问题，并使这种系统适用于大血管(动脉和静脉)血栓研究，以下拟议的研究将开发一种低成本的成像系统，用于对正在发展的血栓进行连续的定量评估。老鼠的大动脉和大静脉(直径最大可达1毫米)将使用具有视频显影功能的微光数码相机进行成像。血栓靶标将用荧光素标记(附着在系统循环的血小板或纤维蛋白原上，或凝血级联酶的抗体上)，以允许监测合并到标准诱导的血栓中(使用PI开发和发布的血栓形成模型)。低成本的激发光源(如发光二极管或激光指示器)将与每个荧光团的激发波长相匹配，使用带通滤光片可以捕获每个荧光团的发射波长的图像。该系统将经过严格的体外测试，然后进行体内分析，直接将结果与现有的血栓定量方法进行比较。这一发现将证明开发这种系统的可行性，并表明其在实验性血栓形成研究中的适用性。这种成像方法可能具有未来适应临床血栓成像的能力。

DESCRIPTION (provided by applicant): Clinical thrombosis has a series and wide-ranging impact on the health of millions of patients, as occurs with heart attacks, strokes, venous thrombo-embolism and disseminated intravascular coagulation. The experimental study of thrombosis requires a consideration of many converging elements within a single physical locale - an intravascular thrombus. The interplay of coagulation factors and platelet activation/accumulation is incompletely understood, as well as the relative involvement of each element in arterial or venous or microvascular thrombosis - distinct entities with different mechanisms of thrombus development and resolution. To overcome the high cost and labor-intensive aspects of recently developed systems for thrombosis study, and to adapt such a system to large-vessel (both artery and vein) thrombosis investigations, the following proposed study will develop a low-cost imaging system for continuous quantitative evaluation of a developing thrombus. Murine large arteries and veins (up to 1 mm in diameter) will be imaged with a low-light digital camera with video development capabilities. Thrombus targets will be labeled with fluorophores (attached to systemically circulating platelets or fibrinogen, or antibodies to coagulation cascade enzymes) to permit monitoring of incorporation into a standardly induced thrombus (using thrombosis models developed and published by the PI). Low-cost excitation light sources (e.g., LEDs or laser pointers) will be matched to the excitation wavelength of each fluorophore, using band-pass filters to permit image capture of emission wavelengths for each fluorophore. The system will undergo rigorous ex vivo testing, followed by in vivo analyses, directly comparing outcomes with established methods for thrombus quantitation. The findings will demonstrate the feasibility of developing such a system and show its applicability to the investigation of experimental thrombosis. This imaging methodology may have a capacity for future adaptation to clinical thrombosis imaging.