Resolution of oblique-plane images in sectioning microscopy.
Resolution of oblique-plane images in sectioning microscopy.
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DOI：
10.1364/oe.19.002662
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发表时间：
2011-01
期刊：
影响因子：
3.8
通讯作者：
Christopher W Smith;E. Botcherby;Tony Wilson
中科院分区：
文献类型：
--
作者：
Christopher W Smith;E. Botcherby;Tony Wilson
活的生物标本在所有三个维度上都表现出微尺度上的时变行为。虽然扫描共聚焦和双光子显微镜能够通过这些标本记录三维图像堆栈，但它们以相对较低的速度这样做，这限制了可以观察到的生物过程的时间分辨率。提高数据采集速率的一种方法是只对感兴趣的样本区域进行成像，因此研究人员最近开始采集倾斜平面或表面的二维图像，这些平面或表面显著延伸到z方向。由于分辨率在x、y和z方向上不均匀，因此图像具有非各向同性分辨率。我们探索这一理论，并表明，图像的倾斜平面可能包含光谱内容，可能不会产生的标本功能完全躺在平面内，而是必须从另一个方向的空间变化。在某些情况下，我们发现图像包含的频率比平面内特征的分辨率极限高三倍。我们证实了这一发现，通过数值模拟和实验上的一种新的，平面成像系统，并建议小心采取这样的图像的解释。
Live biological specimens exhibit time-varying behavior on the microscale in all three dimensions. Although scanning confocal and two-photon microscopes are able to record three-dimensional image stacks through these specimens, they do so at relatively low speeds which limits the time resolution of the biological processes that can be observed. One way to improve the data acquisition rate is to image only the regions of a specimen that are of interest and so researchers have recently begun to acquire two-dimensional images of inclined planes or surfaces extending significantly into the z-direction. As the resolution is not uniform in x, y and z, the images possess non-isotropic resolution. We explore this theoretically and show that images of an oblique plane may contain spectral content that could not have been generated by specimen features lying wholly within the plane but must instead arise from a spatial variation in another direction. In some cases we find that the image contains frequencies three times higher than the resolution limit for in-plane features. We confirm this finding through numerical simulations and experiments on a novel, oblique-plane imaging system and suggest that care be taken in the interpretation of such images.