Orientation imaging of single molecules by wide-field epifluorescence microscopy
Orientation imaging of single molecules by wide-field epifluorescence microscopy
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DOI：
10.1364/josab.20.000554
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发表时间：
2003-03-01
影响因子：
1.9
通讯作者：
Enderlein, J
中科院分区：
文献类型：
--
作者：
Böhmer, M;Enderlein, J
提出了一种直接确定单分子取向的简单成像方法，该方法使用了广视场荧光显微镜和灵敏的CCD相机。成像是在光学器件轻微散焦的情况下进行的，允许基于散焦图像的特征强度分布直接确定单分子取向。给出了这些散焦图像的精确波光计算，并与测量结果吻合较好。这些计算代表了作者所知的第一个完整的偶极发射器在界面上散焦成像的波光模型；考虑了界面上偶极发射的特性以及这种发射和用大数值孔径物镜成像的矢量效应。(C)2003年美国光学学会。
A simple imaging method for direct determination of single-molecule orientations is presented that uses a wide-field epifluorescence microscope and a sensitive CCD camera. Imaging is performed with slight defocusing of the optics, allowing for direct determination of single-molecule orientation based on the characteristic intensity distribution of the defocused images. Exact wave-optical calculations of these defocused images are presented and are in good agreement with the measurements. These calculations represent what is to the authors' knowledge the first complete wave-optical modeling of defocused imaging of dipole emitters at an interface; the peculiarities of dipole emission at an interface and the vector effects of that emission and of imaging with a high-numerical-aperture objective are taken into account. (C) 2003 Optical Society of America.