Quantitative chemical imaging and unsupervised analysis using hyperspectral coherent anti-Stokes Raman scattering microscopy.
Quantitative chemical imaging and unsupervised analysis using hyperspectral coherent anti-Stokes Raman scattering microscopy.
复制标题
使用高光谱相干抗螺旋体拉曼散射显微镜的定量化学成像和无监督分析。
DOI：
10.1021/ac402303g
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发表时间：
2013-11-19
影响因子：
7.4
通讯作者：
Langbein, Wolfgang
中科院分区：
文献类型：
--
作者：
Masia, Francesco;Glen, Adam;Stephens, Phil;Borri, Paola;Langbein, Wolfgang
在这项工作中，我们报告了一种方法来获取和分析高光谱相干反斯托克斯拉曼散射（汽车）显微图像的有机材料和生物样品，从而在一个公正的定量化学分析。该方法采用奇异值分解的汽车强度的平方根，提供了一个自动确定的组件以上的噪声，这是保留。通过两种方法利用磁化率的因果关系从汽车强度谱中提取了在化学组成上呈线性的复杂汽车磁化率谱，并通过与拉曼谱的比较评价了它们的性能。我们使用非负矩阵分解适用于虚部和非共振的真实的部分的磁化率与一个额外的浓度约束，以获得独立变化的化学成分和它们的绝对浓度的绝对磁化率谱。我们证明了该方法提供定量化学分析已知的脂质混合物的能力。然后，我们通过成像富含脂质的干细胞衍生的小鼠脂肪细胞以及具有低密度脂质的分化的胚胎干细胞来显示该方法的相关性。我们检索和可视化最重要的化学成分与水，脂质和蛋白质的光谱分割成细胞周围，脂滴，胞质溶胶和细胞核的图像，我们揭示了细胞的化学结构，通过投影的化学对比度可视化到一些相关的通道的细节。
In this work, we report a method to acquire and analyze hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy images of organic materials and biological samples resulting in an unbiased quantitative chemical analysis. The method employs singular value decomposition on the square root of the CARS intensity, providing an automatic determination of the components above noise, which are retained. Complex CARS susceptibility spectra, which are linear in the chemical composition, are retrieved from the CARS intensity spectra using the causality of the susceptibility by two methods, and their performance is evaluated by comparison with Raman spectra. We use non-negative matrix factorization applied to the imaginary part and the nonresonant real part of the susceptibility with an additional concentration constraint to obtain absolute susceptibility spectra of independently varying chemical components and their absolute concentration. We demonstrate the ability of the method to provide quantitative chemical analysis on known lipid mixtures. We then show the relevance of the method by imaging lipid-rich stem-cell-derived mouse adipocytes as well as differentiated embryonic stem cells with a low density of lipids. We retrieve and visualize the most significant chemical components with spectra given by water, lipid, and proteins segmenting the image into the cell surrounding, lipid droplets, cytosol, and the nucleus, and we reveal the chemical structure of the cells, with details visualized by the projection of the chemical contrast into a few relevant channels.
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影响因子：
7.4
作者：
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通讯作者：
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