Imaging fluorescence from cells in 5-dimensions: x, y, wavelength, lifetime, and anisotropy.

荧光探针被广泛用于研究生物分子在细胞中的位置和功能。使用目前的荧光方法，可以研究细胞内分子的行为。例子包括：聚集、旋转运动和分子间距离。通过测量荧光寿命和各向异性对这三个例子进行了研究。同时监测细胞中多种类型分子在扩展区域内的行为是一个重要的尚未解决的物理问题。这项建议旨在开发、研究和使用一种结合光谱学、荧光寿命和各向异性的荧光成像新方法，目的是同时研究细胞中多种类型的分子的行为。这将产生具有x和y空间维度以及荧光发射波长、荧光寿命和荧光各向异性的图像。这项研究包括三个部分：新仪器的构建，对溶液中确定的荧光化合物混合物的测试，以及对一种重要的细胞表面受体的研究。这项研究的成功完成将为生物学家提供新的方法来研究显微镜领域中多个分子物种的聚集、运动和分子间距离。

Fluorescent probes are widely used to study the location and function of biomolecules in cells. Using current fluorescence methods, the behavior of molecules within cells can be studied. Examples include: clustering, rotational motion, and intermolecular distance. These three examples are studied by measuring fluorescence lifetime and anisotropy. Simultaneous monitoring of the behavior of many types of molecules in cells over an extended area is an important unsolved physical problem. This proposal seeks to develop, study, and use a new approach to fluorescence imaging combining spectroscopy, fluorescence lifetime, and anisotropy with a goal of studying the behavior of many types of molecules in cells simultaneously. This will produce images with x and y spatial dimensions and fluorescence emission wavelength, fluorescence lifetime, and fluorescence anisotropy. The research has three parts: construction of new instrumentation, tests on defined mixtures of fluorescent compounds in a solution, and study of an important cell surface receptor. Successful completion of the study will provide biologists with new ways to study clustering, motion, and intermolecular distances of multiple molecular species in the field of a microscope.