3-D PSF fitting for fluorescence microscopy: implementation and localization application
3-D PSF fitting for fluorescence microscopy: implementation and localization application
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DOI：
10.1111/j.1365-2818.2012.03675.x
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发表时间：
2013-01-01
影响因子：
2
通讯作者：
Unser, M.
中科院分区：
文献类型：
--
作者：
Kirshner, H.;Aguet, F.;Unser, M.
定位显微镜依赖于计算高效的高斯近似的点扩散函数的荧光团位置的计算。理论预测表明，在特定的实验条件下，定位精度显着提高时，使用更真实的模型进行定位。在这里，我们将展示如何通过考虑宽视场显微镜的三维（3-D）点扩散函数模型来实现这一点。我们引入了一个最小二乘点扩散函数拟合框架，利用吉布森和兰尼模型，并提出了一种计算效率高的方法来评估其衍生功能。我们证明了所提出的方法与粒子定位和散焦估计算法的有用性，两者都作为ImageJ的插件实现。
Localization microscopy relies on computationally efficient Gaussian approximations of the point spread function for the calculation of fluorophore positions. Theoretical predictions show that under specific experimental conditions, localization accuracy is significantly improved when the localization is performed using a more realistic model. Here, we show how this can be achieved by considering three-dimensional (3-D) point spread function models for the wide field microscope. We introduce a least-squares point spread function fitting framework that utilizes the Gibson and Lanni model and propose a computationally efficient way for evaluating its derivative functions. We demonstrate the usefulness of the proposed approach with algorithms for particle localization and defocus estimation, both implemented as plugins for ImageJ.