Fiber array based hyperspectral Raman imaging for chemical selective analysis of malaria-infected red blood cells
Fiber array based hyperspectral Raman imaging for chemical selective analysis of malaria-infected red blood cells
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DOI：
10.1016/j.aca.2015.08.025
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发表时间：
2015-09-24
影响因子：
6.2
通讯作者：
Frosch, Torsten
中科院分区：
文献类型：
--
作者：
Brueckner, Michael;Becker, Katja;Frosch, Torsten
提出了一种新的拉曼光谱广场成像装置。它结合了基于光纤阵列的光谱转换器和定制的激光照明系统的优点，可以对敏感的生物样品进行高质量的拉曼化学成像，照明光束的高斯型光强分布由正方形光纤多模光纤整形为光强分布非常均匀的顶帽轮廓，满足克勒条件。30米长的光纤和额外的振荡器有效地破坏了照明光的偏振和相干性。这种均匀的、非相干的照明是复杂生物样品稳定定量成像的基本前提。光纤阵列将拉曼杂散光的二维横向信息转换成具有非常高对比度的分离光谱通道。拉曼图像可以与样品的对应白光显微图像相关联。新的装置能够以非常好的光谱分辨率同时量化整个空间区域的所有拉曼光谱，因此优于其他基于扫描和可调谐滤光片的拉曼成像方法。该装置独特的快速、温和、灵敏和选择性的生物样品化学成像能力被应用于自动化血球蛋白分析。开发了一种特殊的算法，可以根据血球蛋白在红细胞中的分布来生成拉曼图像，而不受其他拉曼散射的影响。新的成像装置与稳健算法相结合，为化学选择性分析恶性疟原虫感染红细胞早期环阶段的疟疾色素血吸虫提供了一种新的、优雅的方法。(C)2015爱思唯尔B.V.保留所有权利。
A new setup for Raman spectroscopic wide-field imaging is presented. It combines the advantages of a fiber array based spectral translator with a tailor-made laser illumination system for high-quality Raman chemical imaging of sensitive biological samples.The Gaussian-like intensity distribution of the illuminating laser beam is shaped by a square-core optical multimode fiber to a top-hat profile with very homogeneous intensity distribution to fulfill the conditions of Koehler. The 30 m long optical fiber and an additional vibrator efficiently destroy the polarization and coherence of the illuminating light. This homogeneous, incoherent illumination is an essential prerequisite for stable quantitative imaging of complex biological samples. The fiber array translates the two-dimensional lateral information of the Raman stray light into separated spectral channels with very high contrast. The Raman image can be correlated with a corresponding white light microscopic image of the sample. The new setup enables simultaneous quantification of all Raman spectra across the whole spatial area with very good spectral resolution and thus outperforms other Raman imaging approaches based on scanning and tunable filters.The unique capabilities of the setup for fast, gentle, sensitive, and selective chemical imaging of biological samples were applied for automated hemozoin analysis. A special algorithm was developed to generate Raman images based on the hemozoin distribution in red blood cells without any influence from other Raman scattering. The new imaging setup in combination with the robust algorithm provides a novel, elegant way for chemical selective analysis of the malaria pigment hemozoin in early ring stages of Plasmodium falciparum infected erythrocytes. (C) 2015 Elsevier B.V. All rights reserved.