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使用全内反射荧光法(TIRF)测定吸附荧光团的取向分布。I. 理论

Determination of the orientation distribution of adsorbed fluorophores using TIRF. I. Theory.

作者信息

Bos M A, Kleijn J M

机构信息

Department of Physical and Colloid Chemistry, Agricultural University, Wageningen, The Netherlands.

出版信息

Biophys J. 1995 Jun;68(6):2566-72. doi: 10.1016/S0006-3495(95)80439-X.

Abstract

The spectroscopic technique total internal reflection fluorescence can be used for determination of the orientation of adsorbed fluorescent molecules. The underlying theory is presented in general terms and elaborated in detail for the case that the fluorescent group is a porphyrin ring. It is shown that order parameters of the orientation distribution can be obtained if both the fluorescence intensity and its polarization are measured as functions of the polarization of the incident laser beam. From these order parameters an approximation of the orientation distribution can be derived by the maximum-entropy method.

摘要

全内反射荧光光谱技术可用于测定吸附荧光分子的取向。文中概述了其基本理论,并针对荧光基团为卟啉环的情况进行了详细阐述。结果表明,如果同时测量荧光强度及其偏振度随入射激光束偏振度的变化,就可以得到取向分布的序参量。通过最大熵方法,可从这些序参量推导出取向分布的近似值。

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