Improved quantification of collagen anisotropy with polarization-resolved second harmonic generation microscopy
Improved quantification of collagen anisotropy with polarization-resolved second harmonic generation microscopy
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DOI：
10.1002/jbio.201600197
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发表时间：
2017-09-01
影响因子：
2.8
通讯作者：
Stanciu, George A.
中科院分区：
文献类型：
--
作者：
Hristu, Radu;Stanciu, Stefan G.;Stanciu, George A.
成像组织样品的偏振分辨二次谐波产生显微镜提供了定性和定量的见解，以无标签的方式胶原组织。偏振分辨二次谐波产生显微镜通过增加激光束偏振分量和应用不同的定量指标（如各向异性因子），超越了简单的基于强度的成像。因此，它提供了胶原蛋白排列的宝贵信息，而不是单独的强度测量。目前建立的方法仅限于计算特定激光束偏振的各向异性因子，并且尚未确定如何选择最佳激光束偏振的一般准则。在这里，我们介绍了一种新的方法来选择最佳的激光束偏振来表征组织，利用各向异性来识别癌症特征。我们发现，各向异性因子对二次谐波强度表现出类似的光束偏振依赖性，并将其与记录图像的快速傅里叶变换分析计算的胶原取向指数相结合，建立了一个框架，用于选择最优的激光束偏振，以准确解释偏振分辨二次谐波生成显微镜图像和各向异性图。从而更好地区分健康区域和发育不良区域。[图形]皮肤组织的SHG图像(a)和选定的感兴趣区域，我们计算SHG强度(b)和各向异性因子(c)依赖于激光束偏振和FFT谱(d)来评估胶原取向指数。
Imaging tissue samples by polarization-resolved second harmonic generation microscopy provides both qualitative and quantitative insights into collagen organization in a label-free manner. Polarization-resolved second harmonic generation microscopy goes beyond simple intensity-based imaging by adding the laser beam polarization component and applying different quantitative metrics such as the anisotropy factor. It thus provides valuable information on collagen arrangement not available with intensity measurements alone. Current established approaches are limited to calculating the anisotropy factor for only a particular laser beam polarization and no general guidelines on how to select the best laser beam polarization have yet been defined. Here, we introduce a novel methodology for selecting the optimal laser beam polarization for characterizing tissues using the anisotropy in the purpose of identifying cancer signatures. We show that the anisotropy factor exhibits a similar laser beam polarization dependence to the second harmonic intensity and we combine it with the collagen orientation index computed by Fast Fourier Transform analysis of the recorded images to establish a framework for choosing the laser beam polarization that is optimal for an accurate interpretation of polarization-resolved second harmonic generation microscopy images and anisotropy maps, and hence a better differentiation between healthy and dysplastic areas.[GRAPHICS]SHG image of skin tissue (a) and a selected area of interest for which we compute the SHG intensity (b) and anisotropy factor (c) dependence on the laser beam polarization and also the FFT spectrum (d) to evaluate the collagen orientation index.