Antibody Imaging for Regional Flow Measurements

本研究涉及开发高分辨率确定组织和器官区域血流的程序。目前的区域流量测定方法受到微球大小和计数统计的限制。本研究验证了一个假设，即荧光标记抗体作为“分子微球”，将与微区域流动成比例地附着在毛细血管结合位点上。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来测量整个器官的微区域血流。该项目将为生理/医学研究人员提供一种独特的工具，用于高分辨率量化区域血流。I期研究将证明荧光抗体作为大鼠心脏分子血流标记的能力。第二阶段将把这种方法扩展到不同颜色的荧光染料附着在抗体上的研究和针对毛细血管内皮的不同抗体的研究。第二阶段还将展示该方法在不同器官和大型动物中流量测量的实用性，并将开发高分辨率流量显示软件。这项技术创新将推动成像冷冻微生物组/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（降低组织体积元素尺寸）可以极大地扩展使用该技术的研究类型，并增加对细胞细胞组成像的需求。

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.