Axial tomographic confocal fluorescence microscopy
Axial tomographic confocal fluorescence microscopy
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DOI：
10.1046/j.1365-2818.2002.01000.x
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发表时间：
2002-04-01
期刊：
影响因子：
--
通讯作者：
Cremer, C
中科院分区：
文献类型：
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作者：
Heintzmann, R;Cremer, C
通过样品的物理旋转，轴向断层扫描可以从对象中获取否则无法获取的空间信息。该方法与共聚焦显微镜相结合，可以从根本上提高有效三维分辨率。在本报告中，我们提出了一种高分辨率重建共焦轴向层析成像数据的新方法。该方法自动确定相对旋转角度，对不同旋转视图的数据进行对齐，并重建单个高分辨率3D数据集。重建利用已知的点扩展函数，并基于从多个（在我们的情况下是三个）角度视图同时执行的无约束最大似然反卷积。它被应用于模拟和实验共聚焦数据集。对分辨率增益进行了量化，并研究了超松弛因子的选择对收敛速度的影响。与单角度共焦数据重建相比，轴向层析成像和重建获得了明显提高的三维分辨率。
By physical rotation of the sample, axial tomography enables the acquisition of otherwise inaccessible spatial information from an object. In combination with confocal microscopy, the method can fundamentally improve the effective three-dimensional (3D) resolution. In this report we present a novel method for high resolution reconstruction of confocal axial tomographic data. The method automatically determines the relative angles of rotation, aligns the data from different rotational views and reconstructs a single high resolution 3D dataset. The reconstruction makes use of a known point spread function and is based on an unconstrained maximum likelihood deconvolution performed simultaneously from multiple (in our case three) angular views. It was applied to simulated as well as to experimental confocal datasets. The gain in resolution was quantified and the effect of choice of overrelaxation factors on the speed of convergence was investigated. A clearly improved 3D resolution was obtained by axial tomography together with reconstruction as compared with reconstruction of confocal data from only it single angular view.