Antibody Imaging for Regional Flow Measurements

描述(由申请人提供)： 这项研究涉及开发一种使用自动化仪器程序来量化组织中区域血流的方法。I期结果表明，在器官连续切片过程中，针对毛细血管内皮细胞抗原的荧光抗体可以在从冰冻组织块记录的连续荧光图像中进行空间定位。通过使用分子作为流动标记(分子微球)，消除了对荧光和放射性微球方法施加的统计限制，极大地扩展了器官流动图的细节。在第一阶段，荧光抗体被鉴定为在心脏血管系统中具有良好的保留性和区域沉积，这与同时注射的15微米直径的荧光微球密切相关。验证了分子微球方法的有效性和成像方法的可行性。第二阶段将集中在心脏的体内研究，开发分析程序，并由合作者进行肺和脑的验证研究。还将开发在同一器官中进行多个流量测量的方法。该项目将为研究人员提供方法和仪器，以测量比目前技术可能的组织体积小得多的区域血流。

DESCRIPTION (provided by applicant): This research involves the development of a method to quantify regional flow in tissues using automated instrumental procedures. Phase I results have demonstrated that fluorescent antibodies, specific to antigens in capillary endothelium, can be spatially located in sequential fluorescence images recorded from frozen tissue blocks during organ serial sectioning. Statistical limitations imposed on fluorescent and radioactive microsphere methods are removed by the use of molecules as flow markers (molecular microspheres), greatly expanding the details of organ-flow maps. During Phase I, fluorescent antibodies were identified that have excellent retention in heart vascular systems and regional deposition which correlates strongly with simultaneously injected 15 micron-diameter flourescent microspheres. Both the validity of the molecular microsphere approach and the feasibility of the imaging methodology have been demonstrated. Phase II will concentrate on in vivo studies in heart, development of analysis procedures and validation studies in lung and brain to be undertaken by collaborators. Methods will also be developed to conduct multiple flow measurements in the same organ. This project will provide researchers with methods and instrumentation to measure regional flow in tissue volumes much smaller than those possible with current techniques.