FLUORESCENCE IMAGING OF REGIONAL FLOW IN HEART

这项研究涉及开发全自动仪器程序来评估组织中的微区域循环。第一阶段的结果表明，在器官连续切片过程中，从冰冻组织块记录的连续荧光图像中，可以在空间上定位细胞特定的荧光染料或“分子微球”。通过使用分子大小的微球，消除了对当前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谱的细节。在第一阶段，几种染料已被确认为在心脏血管系统中具有良好的保留力，它们的区域沉积与同时注射的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。第二阶段将集中于调整和优化微区域血流成像的成像系统，并将通过与华盛顿大学荧光微球资源中心的合作对该技术进行严格的验证。该项目将为研究人员提供快速、自动地评估组织体积小于当前技术可能规模的1/1000的微区域血流的仪器，同时使用与放射性和荧光微球血流测定所涉及的标准程序类似的程序。拟议的商业应用：快速和自动评估组织中分子微球分布的仪器和方法将在从事生理学和循环研究的实验室中得到应用。提案中分子微球方法学的发展将极大地促进巴洛科学微球成像系统在这些实验室的商业市场。

This research involves the development of fully-automated instrumentation procedures to evaluate microregional circulation in tissues. Phase I results have demonstrated that cell-permeant fluorescent dyes or "molecular microspheres" can be spatially located in sequential fluorescence images recorded from frozen tissue blocks during organ serial sectioning. Statistical limitations imposed on current fluorescent and radioactive microsphere methods are removed by the use of molecule-sized microspheres, expanding greatly the details of organ-flow mapping. During Phase I, several dyes have been identified as having excellent retention in heart vascular systems and their regional deposition correlates strongly with simultaneously injected 15-micron diameter fluorescent microspheres. Both the validity of the molecular microsphere approach and the feasibility of the imaging methodology have been demonstrated. Phase II will concentrate on adapting and optimizing the imaging system for microregional flow imaging and will involve a rigorous validation of the technique through collaboration with the Fluorescent Microsphere Resource Center at the University of Washington. This project will provide researchers with instrumentation for rapid, automatic evaluation of microregional flow in tissue volumes less than 1/1000 the size of those possible with current techniques, while using procedures similar to the standard procedures involved in radioactive and fluorescent microsphere flow determinations. PROPOSED COMMERCIAL APPLICATIONS: Instrumentation and methods to rapidly and automatically evaluate molecular microsphere distributions in tissue will find applications in laboratories engaged in physiology and circulation research. The development of molecular microsphere methodology in the proposal will significantly enhance the commercial market for the Barlow Scientific microsphere imaging system in such laboratories.