Blind deconvolution of 3D transmitted light brightfield micrographs
Blind deconvolution of 3D transmitted light brightfield micrographs
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DOI：
10.1046/j.1365-2818.2000.00751.x
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发表时间：
2000-11-01
影响因子：
2
通讯作者：
O'Connor, NJ
中科院分区：
文献类型：
--
作者：
Holmes, TJ;O'Connor, NJ
用示例图像总结了先前公布的用于3D透射光明场（TLB）显微术的盲解卷积算法（Holmes等人，Handbook of Biological Confocal Microscopy（1995））。本文的主要重点是更彻底地讨论这种方法的重要性和实用性，并提供更详细的证据，有些是定量的，它的必要性。制备辣根过氧化物酶（HRP）染色的锥体神经元样品，并评价其清晰观察精细结构（包括树突和棘）的能力。它表明，精细的脊柱结构的外观，并确定脊柱类别的手段，是可能的，通过使用盲反卷积。来自反射光共聚焦显微镜的相同样品的图像的比较，这是观察这些HRP染色样品的3D结构的常规正确显微镜方式，表明盲去卷积方法对于清楚地显示具有更少失真和更好的棘分辨率的结构而言是优越得多的上级。这项研究的主要意义在于，现在可以通过吸收染料的光学显微镜获得清晰的3D结构图像。这是重要的，因为TLB显微镜可能是生命科学实验室中最广泛使用的模态，然而，直到现在还没有可靠的手段来提供3D结构的清晰可视化。盲去卷积方法的主要重要性在于，它消除了测量光学系统的点扩散函数的需要，因此现在提供一种能够被显微镜工作者普遍使用的3D光学显微镜去卷积方法变得现实。
The blind deconvolution algorithm for 3D transmitted light brightfield (TLB) microscopy, published previously (Holmes et al. Handbook of Biological Confocal Microscopy (1995), is summarized with example images. The main emphasis of this paper is to discuss more thoroughly the importance and usefulness of this method and to provide more detailed evidence, some being quantitative, of its necessity. Samples of horseradish peroxidase (HRP)-stained pyramidal neurones were prepared and evaluated for the ability to see fine structures clearly including the dendrites and spines. It is demonstrated that the appearance of fine spine structure, and means of identifying spine categories, is made possible by using blind deconvolution. A comparison of images of the same sample from reflected light confocal microscopy, which is the conventional right microscopic way of viewing the 3D structure of these HRP-stained samples, shows that the blind deconvolution method is far superior for clearly showing the structure with less distortion and better resolution of the spines. The main significance of this research is that it is now possible to obtain clear images of 3D structure by light microscopy of absorbing stains. This is important because the TLB microscope is probably the most widely used modality in the life-science laboratory, yet, until now there has been no reliable means for it to provide visualization of 3D structure clearly The main importance of the blind deconvolution approach is that it obviates the need to measure the point spread function of the optical system, so that it now becomes realistic to provide a 3D light microscopic deconvolution method that can be pervasively used by microscopists.