Antibody Imaging for Regional Flow Measurements

这项研究涉及开发程序，以确定高分辨率组织和器官中的区域流动。目前的区域流量测定方法受到微球尺寸和计数统计的限制。这项研究测试的假设，荧光标记的抗体，作为“分子微球”，将附着到毛细血管结合位点的比例微区域流量。与目前基于微球的测量相比，来自结合染料分子的荧光强度可以潜在地将区域流量测量的体积分辨率提高1，000至1，000，000倍。该项目将开发荧光图像分析方法，以测量整个器官中的微区域流量。该项目将为生理/医学研究人员提供一种独特的工具，用于以高分辨率量化区域流量。第一阶段研究将证明荧光抗体作为大鼠心脏分子流标记物的能力。第二阶段将扩大这一方法的研究不同颜色的荧光染料连接到抗体和不同的抗体特异性毛细血管内皮。第二阶段还将展示该方法在不同器官和大型动物中的流量测量的实用性，并将开发高分辨率流量显示软件。这一技术创新将推动成像冷冻切片机/TM的发展，以进行组织和器官的综合生理研究。拟议的商业应用：潜在的市场包括目前进行低分辨率流量测量的数百个实验室，以及希望在实验动物上验证MRI方法的研究人员。提高分辨率（降低组织体积元素大小）可以大大扩展使用该技术的研究类型，并增加对细胞切片机成像的需求。

This research involves developing procedures to determine regional flow in tissues and organs at high resolution. Current methods for regional flow determinations are limited by microsphere size and counting statistics. This study tests the hypothesis that fluorescently-labeled antibodies, serving as "molecular microspheres", will attach to capillary binding sites in proportion to microregional flow. As compared to current microsphere-based measurements, the fluorescence intensity from bound dye molecules could potentially improve volume resolution of regional flow measurements by factors of 1,000 to 1,000,000. The project will develop fluorescence image analysis methods to measure microregional flow in whole organs. This project will provide physiological/medical researchers with a unique tool for quantifying regional flow at high resolution.. Phase I research will demonstrate the ability of fluorescent antibodies to serve as molecular flow markers in rat heart. Phase II will extend this methodology to studies of different colored fluorochromes attached to antibodies and to different antibodies specific to capillary endothelium. Phase II will also demonstrate the utility of the method for flow measurements in different organs and large animals and will develop high resolution flow display software. This technological innovation will advance the development of the Imaging CryoMicrotome/TM to perform integrative physiological studies in tissues and organs. PROPOSED COMMERCIAL APPLICATIONS: Potential markets include the hundreds of laboratories that currently do low resolution flow measurements, as well as researchers wishing to validate MRI methods on experimental animals. Increasing the resolution (decreasing tissue volume element size) could greatly expand the types of studies using this technology and increase demand for cytomicrotome imaging.