Harmonic microscopy of isotropic and anisotropic microstructure of the human cornea
Harmonic microscopy of isotropic and anisotropic microstructure of the human cornea
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DOI：
10.1364/oe.18.005028
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发表时间：
2010-03-01
期刊：
影响因子：
3.8
通讯作者：
Beaurepaire, Emmanuel
中科院分区：
文献类型：
--
作者：
Olivier, Nicolas;Aptel, Florent;Beaurepaire, Emmanuel
在这项研究中，我们展示了完整人角膜的三次谐波（THG）和二次谐波（SHG）联合显微镜图像，并从实验和理论上分析了THG信号的来源。多谐显微镜提供了详细的图像的角膜微观结构在其整个厚度。THG信号的一个成分来源于细胞结构，另一个成分来源于基质中连续胶原片之间的各向异性变化。这种各向异性相关的信号可以通过圆形入射偏振来检测，并提供基质薄片的堆叠和组织尺度非均质性的对比图像。前向辐射THG和SHG信号通常是反相关的，表明最大THG来自片层界面，而最大SHG来自片层内部。偏振分辨THG成像反映了片层的互向各向异性。我们提出了层状各向异性样品的THG成像模型和数值计算，以解释我们的观察结果。(C) 2010年美国光学学会
In this study we present combined third-harmonic generation (THG) and second-harmonic generation (SHG) microscopy images of intact human corneas, and we analyze experimentally and theoretically the origin of the THG signal. Multiharmonic microscopy provides detailed images of the cornea microstructure over its entire thickness. A component of the THG signal originates from cellular structures and another one originates from anisotropy changes between successive collagen lamellae in the stroma. This anisotropy-related signal can be specifically detected using circular incident polarization, and provide contrasted images of the stacking and tissue-scale heterogeneity of stromal lamellae. Forward-radiated THG and SHG signals are generally anticorrelated, indicating that maximum THG is obtained from lamellar interfaces whereas maximum SHG is obtained from within lamellae. Polarization-resolved THG imaging reflects the alternate anisotropy directions of the lamellae. We present a model for THG imaging of layered anisotropic samples and numerical calculations that account for our observations. (C) 2010 Optical Society of America