Low-Cost Digital Imaging System for Evaluating Thrombosis

描述（由申请人提供）：临床血栓形成对数百万患者的健康有一系列广泛的影响，如心脏病发作、中风、静脉血栓栓塞和弥散性血管内凝血。血栓形成的实验研究需要考虑在一个单一的物理位置-血管内血栓内的许多聚合元素。凝血因子和血小板活化/积聚的相互作用以及动脉或静脉或微血管血栓形成中每种元素的相对参与尚不完全清楚-这些不同的实体具有不同的血栓形成和消退机制。为了克服最近开发的用于血栓形成研究的系统的高成本和劳动密集型方面，并且为了使这样的系统适应于大血管（动脉和静脉两者）血栓形成研究，以下提议的研究将开发用于持续定量评价正在形成的血栓的低成本成像系统。将使用具有视频显影能力的低光数码相机对小鼠大动脉和静脉（直径达1 mm）进行成像。血栓靶点将用荧光团标记（附着于全身循环血小板或纤维蛋白原，或凝血级联酶的抗体），以允许监测掺入标准品诱导的血栓（使用PI开发和发表的血栓形成模型）。低成本激发光源（例如，LED或激光指示器）将与每个荧光团的激发波长匹配，使用带通滤波器以允许每个荧光团的发射波长的图像捕获。该系统将进行严格的体外测试，然后进行体内分析，直接将结果与已建立的血栓定量方法进行比较。研究结果将证明开发这样一个系统的可行性，并显示其适用于实验性血栓形成的调查。这种成像方法可能有能力在未来适应临床血栓成像。

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.