全内反射荧光显微术应用于单分子荧光的纵向成像

利用这种显微术中激发场的纵向强度呈指数衰减的分布特性，测得的荧光强 度将强烈依赖渗透深度（强度衰减到1/e时的距离），从而快速直接的确定单分子荧光 团间的纵向间隔、每个荧光团的纵向绝对位置和荧光团的半径大小，即实现荧光分子三维分 布的重构.在整个重构的过程中，只需要改变一次入射角的大小，即只需探测两幅荧光分子 的全内反射成像. 关键词： 全内反射荧光显微术 纵向重构

The application of total internal reflection fluorescence microscopy in single fluorophore molecules axial imaging

Here we present a method of modeling fluorophore molecules as spheres in evanescent fields mathematically instead of the assumption of top-hat function distribution. Based on this model, we discuss that it is possible to calculate relative separation distances of the molecules, and their axial positions and diameters with the total internal reflection fluorescence microscopy(TIRFM) due to its unique intensity pattern in the z direction. In the whole process, we need merely to change the incident angle once, that is to get only two frame imaging of TIRFM.