Quantification of collagen fiber structure using second harmonic generation imaging and two-dimensional discrete Fourier transform analysis: Application to the human optic nerve head
Quantification of collagen fiber structure using second harmonic generation imaging and two-dimensional discrete Fourier transform analysis: Application to the human optic nerve head
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DOI：
10.1002/jbio.201800376
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发表时间：
2019-05-01
影响因子：
2.8
通讯作者：
Boote, Craig
中科院分区：
文献类型：
--
作者：
Pijanka, Jacek K.;Markov, Petar P.;Boote, Craig
二次谐波发生（SHG）显微镜被广泛用于图像胶原纤维微结构，由于其高的空间分辨率，光学切片能力和相对无损的样品制备。SHG图像的定量需要灵敏的方法来捕获纤维对齐。本文提出了一种基于二维离散傅里叶变换（DFT）的方法，从SHG图像中分析胶原纤维结构。该方法包括积分周期性加上平滑图像分解，用于校正DFT边缘不连续伪影，避免了更常用的开窗方法所遇到的外围图像数据的丢失。外展参数包括胶原纤维的取向分布、取向胶原含量和胶原纤维沿主取向沿着分散的程度。我们证明了它的应用，以确定在人类视神经乳头的胶原蛋白微结构，显示其能够准确地捕捉特征性的结构特征，包括径向纤维排列在最内层的边界巩膜和一个圆周胶原环中基质组织。更高的空间分辨率渲染个别筛板梁内的神经头也被证明。该方法的验证提供的形式的相关结果从广角X射线散射和所提出的方法应用到其他纤维组织。
Second harmonic generation (SHG) microscopy is widely used to image collagen fiber microarchitecture due to its high spatial resolution, optical sectioning capabilities and relatively nondestructive sample preparation. Quantification of SHG images requires sensitive methods to capture fiber alignment. This article presents a two-dimensional discrete Fourier transform (DFT)-based method for collagen fiber structure analysis from SHG images. The method includes integrated periodicity plus smooth image decomposition for correction of DFT edge discontinuity artefact, avoiding the loss of peripheral image data encountered with more commonly used windowing methods. Outputted parameters are as follows: the collagen fiber orientation distribution, aligned collagen content and the degree of collagen fiber dispersion along the principal orientation. We demonstrate its application to determine collagen microstructure in the human optic nerve head, showing its capability to accurately capture characteristic structural features including radial fiber alignment in the innermost layers of the bounding sclera and a circumferential collagen ring in the mid-stromal tissue. Higher spatial resolution rendering of individual lamina cribrosa beams within the nerve head is also demonstrated. Validation of the method is provided in the form of correlative results from wide-angle X-ray scattering and application of the presented method to other fibrous tissues.