Antibody Imaging for Regional Flow Measurements

这项研究涉及开发程序，以高分辨率确定组织和器官中的区域流动。目前测定区域流量的方法受到微球大小和计数统计的限制。这项研究验证了一种假设，即作为“分子微球”的荧光标记抗体将与微区域流动成比例地附着在毛细血管结合部位。与目前基于微球的测量相比，结合染料分子的荧光强度可能会将区域流动测量的体积分辨率提高1000到1,000,000倍。该项目将开发荧光图像分析方法来测量整个器官的微区域血流。该项目将为生理学/医学研究人员提供一种独特的工具，以高分辨率量化区域血流。第一阶段的研究将证明荧光抗体在大鼠心脏中作为分子流标记的能力。第二阶段将把这一方法扩展到对不同颜色的荧光染料与抗体和不同的毛细血管内皮细胞特异性抗体的研究。第二阶段还将演示该方法在不同器官和大型动物中的流量测量的实用性，并将开发高分辨率的流量显示软件。这项技术创新将推动成像冷冻微型机/TM的发展，以进行组织和器官的综合生理学研究。拟议的商业应用：潜在市场包括目前进行低分辨率流量测量的数百家实验室，以及希望在实验动物身上验证MRI方法的研究人员。提高分辨率(减小组织体积元素大小)可以极大地扩展使用该技术的研究类型，并增加对细胞显微成像的需求。

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.