Resolution improvement by 3-D reconstructions from tilted views in axial tomography and confocal theta microscopy

Kurt Sätzler, Roland Eils
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引用次数: 20

Abstract

The resolution along the optical axis is much less than the lateral resolution in both confocal and conventional fluorescence light microscopy. To correct for the anisotropy in resolution one may generate tilted views from the same object and subsequently merge these tilted views into one image with improved isotropic resolution. It has been suggested to obtain tilted views in confocal theta microscopy with double observation by illuminating the object from one side and simultaneously detecting the emitted light in the direction parallel to the illumination direction and in a direction with an angle theta to the illumination direction. Alternatively, one might use the usual optical set-up of a light microscope together with a tilting device to rotate the object under the microscope. With this method the object can be viewed from all directions (axial tomography). In this paper we investigate two methods for merging tilted views. If the image sets obtained from the different tilt angles are interpreted as statistic samples of one and the same object, a gain in resolution along the optical axis is accompanied by a significant trade-off in lateral resolution. An optical interpretation of the image formation in each single object point by retaining the frequencies with the highest amplitudes in the different tilted views provides a gain in axial resolution without significant decrease in lateral resolution. We will describe a quantitative method to determine the gain in resolution dependent on the number and geometry of the tilted views. It will be demonstrated that a previously suggested reconstruction method implying a multiplication of tilted views in the spatial domain does not improve resolution. Simulated reconstructions based on measured point spread functions in axial tomography and confocal theta microscopy with double observation will demonstrate the validity of our reconstruction methods.

通过轴向断层扫描和共聚焦theta显微镜中倾斜视图的三维重建提高分辨率
在共聚焦和常规荧光显微镜中,沿光轴的分辨率远小于横向分辨率。为了纠正分辨率的各向异性,可以从同一对象生成倾斜视图,然后将这些倾斜视图合并为具有改进的各向同性分辨率的图像。建议在共聚焦θ显微镜中,通过从物体的一侧照射物体,同时检测与照射方向平行的方向和与照射方向成θ角的方向的发射光,从而获得双观察的倾斜视图。另外,人们可以使用光学显微镜的常规光学装置和一个倾斜装置来旋转显微镜下的物体。用这种方法可以从各个方向观察物体(轴向断层扫描)。本文研究了两种合并倾斜视图的方法。如果从不同倾斜角度获得的图像集被解释为同一物体的统计样本,那么沿着光轴的分辨率增益伴随着横向分辨率的显着权衡。通过保留不同倾斜视图中具有最高振幅的频率,对每个单个目标点的图像形成进行光学解释,可以在不显著降低横向分辨率的情况下获得轴向分辨率。我们将描述一种定量方法来确定依赖于倾斜视图的数量和几何形状的分辨率增益。这将证明,以前建议的重建方法意味着在空间域中倾斜视图的乘法并不能提高分辨率。基于轴向层析成像和共聚焦显微镜中测点扩展函数的模拟重建将证明我们的重建方法的有效性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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