PHOTOSTABLE FLUORESCENT DYES FOR BIOCHEM/MICROSCOPY

我们建议合成新的反应性荧光染料的基础上， 非常耐光的萘酰亚胺类染料。这些新的污渍 将适用于许多相同的用途，因为目前可用的 荧光素、罗丹明、Cy 3、香豆素等标记剂，并将 具有结合大斯托克斯位移、高 量子产率和适用于荧光的适当长的寿命 极化免疫测定 在第一阶段，我们将制备几种萘酰亚胺染料， 用助色剂使它们的激发向红移， 发射波长，并添加一个官能团，可以与 双功能接头以产生反应性荧光探针阵列。 将测试这些探针标记氨基的能力 蛋白质如肌动蛋白和抗生物素蛋白。我们将确定每一个的效果 改变染料的物理常数（波长 激发和发射、消光系数、消光系数，以及 荧光量子产率（phiflu）和测试的光稳定性 染料对常用的荧光标记。我们还将确定 蛋白质的标记程度（#标记/蛋白质分子）和 phiflu的标签上的蛋白质。 第一阶段的工作将设计 以确定建立新的 光化学稳定的标记剂，以取代现有的染料， 具有不太理想的性质。 拟议的商业应用：在某种程度上，我们的新荧光团 改进现有的荧光标记剂，他们将找到一种现成的 市场 由于它们的高光稳定性和高量子产率， 它们将特别用作光的荧光标记物 显微镜应用它们的大斯托克斯位移也会使它们 适用于荧光偏振免疫测定。

We propose to synthesize new reactive fluorescent stains based on an extraordinarily photostable ClaSS of naphthalimide dyes. These new stains will be applicable to many of the same uses as the currently available fluorescein, rhodamine, Cy3, coumarin and other labeling agents, and will have the additional advantage of combining large Stokes shift, high quantum yield, and suitably long lifetime to apply to fluorescence polarization immunoassays. In Phase I we will prepare several naphthalimide dyes, derivatize them with auxochromes to bathochromically shift their excitation and the emission wavelengths, and add a functional group which can react with bifunctional linkers to yield an array of reactive fluorescent probes. These probes will be tested for their ability to label amino groups in proteins such as actin and avidin. We will determine the effect of each modification of the dye on the photophysical constants (wavelength of excitation and emission, extinction coefficient, epsilon, and fluorescence quantum yield, (phiflu) and test the photostability of the dyes against commonly used fluorescent labels. We will also determine the degree of labeling of the proteins (# labels/protein molecule) and the phiflu of the label on the protein. The Phase I work will be designed to determine the feasibility of the approach to creating new photochemically stable labeling agents to replace existing dyes which have less desirable properties. PROPOSED COMMERCIAL APPLICATION: To the extent that our new fluorophores improve on existing fluorescent labeling agents, they will find a ready market. Because of their high photostability and high quantum yield, they will be particularly useful as fluorescent markers for light microscopy applications. Their large Stokes shifts will also make them suitable for fluorescence polarization immunoassays.