Super-resolution orientation estimation and localization of fluorescent dipoles using 3-D steerable filters
Super-resolution orientation estimation and localization of fluorescent dipoles using 3-D steerable filters
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DOI：
10.1364/oe.17.006829
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发表时间：
2009-04-13
期刊：
影响因子：
3.8
通讯作者：
Unser, Michael
中科院分区：
文献类型：
--
作者：
Aguet, Francois;Geissbuehler, Stefan;Unser, Michael
在图像采集期间固定的荧光团产生衍射图案，该衍射图案是荧光团的基础偶极的取向的特征。荧光定位显微镜技术（如PALM和STORM）通过将基于高斯的拟合算法应用于单个荧光团的聚焦图像来实现超分辨率;当应用于固定偶极子时，这可能导致5-20 nm范围内的偏差。我们介绍了一种方法的联合估计的位置和方向的偶极子，基于一个物理上现实的图像形成模型作为一个3-D可操纵滤波器的表示。我们的方法依赖于一个单一的，分散的收购。我们建立理论，基于本地化的分辨率限制估计精度使用Cramer-Rao界限，并通过实验表明，估计精度至少为5纳米的位置和至少2度的方向可以实现。通过将图像形成模型应用于估计的位置/取向对而生成的图案与实验观察结果紧密匹配。(C)2009年美国光学学会
Fluorophores that are fixed during image acquisition produce a diffraction pattern that is characteristic of the orientation of the fluorophore's underlying dipole. Fluorescence localization microscopy techniques such as PALM and STORM achieve super-resolution by applying Gaussian-based fitting algorithms to in-focus images of individual fluorophores; when applied to fixed dipoles, this can lead to a bias in the range of 5-20 nm. We introduce a method for the joint estimation of position and orientation of dipoles, based on the representation of a physically realistic image formation model as a 3-D steerable filter. Our approach relies on a single, defocused acquisition. We establish theoretical, localization-based resolution limits on estimation accuracy using Cramer-Rao bounds, and experimentally show that estimation accuracies of at least 5 nm for position and of at least 2 degrees for orientation can be achieved. Patterns generated by applying the image formation model to estimated position/orientation pairs closely match experimental observations. (C) 2009 Optical Society of America