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Mirrored stainless steel substrate provides improved signal for Raman spectroscopy of tissue and cells

机译:镜面不锈钢基板可改善组织和细胞的拉曼光谱信号

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摘要

Raman spectroscopy (RS) is a powerful technique that permits the non‐destructive chemical analysis of cells and tissues without the need for expensive and complex sample preparation. To date, samples have been routinely mounted onto calcium fluoride (CaF2) as this material possesses the desired mechanical and optical properties for analysis, but CaF2 is both expensive and brittle and this prevents the technique from being routinely adopted. Furthermore, Raman scattering is a weak phenomenon and CaF2 provides no means of increasing signal. For RS to be widely adopted, particularly in the clinical field, it is crucial that spectroscopists identify an alternative, low‐cost substrate capable of providing high spectral signal to noise ratios with good spatial resolution. Results show that these desired properties are attainable when using mirrored stainless steel as a Raman substrate. When compared with CaF2, data show that stainless steel has a low background signal and provides an average signal increase of 1.43 times during tissue analysis and 1.64 times when analyzing cells. This result is attributed to a double‐pass of the laser beam through the sample where the photons from the source laser and the forward scattered Raman signal are backreflected and retroreflected from the mirrored steel surface and focused towards collection optics. The spatial resolution on stainless steel is at least comparable to that on CaF2 and it is not compromised by the reflection of the laser. Steel is a fraction of the cost of CaF2 and the reflection and focusing of photons improve signal to noise ratios permitting more rapid mapping. The low cost of steel coupled with its Raman signal increasing properties and robust durability indicates that steel is an ideal substrate for biological and clinical RS as it possesses key advantages over routinely used CaF2. © 2016 The Authors. Journal of Raman Spectroscopy Published by John Wiley & Sons Ltd.
机译:拉曼光谱法(RS)是一项功能强大的技术,无需进行昂贵且复杂的样品制备,即可对细胞和组织进行无损化学分析。迄今为止,由于该材料具有用于分析的所需机械和光学特性,因此通常已将样品常规安装到氟化钙(CaF2)上,但是CaF2既昂贵又易碎,这妨碍了该技术的常规采用。此外,拉曼散射是微弱的现象,而CaF2无法提供增加信号的手段。为了使RS被广泛采用,特别是在临床领域,光谱学家确定一种替代的低成本底物至关重要,该底物能够提供高光谱信噪比并具有良好的空间分辨率。结果表明,当使用镜面不锈钢作为拉曼底材时,可以获得这些所需的性能。与CaF2相比,数据显示不锈钢的背景信号低,在组织分析过程中平均信号增加1.43倍,在分析细胞过程中平均信号增加1.64倍。该结果归因于激光束两次穿过样品,其中来自源激光器的光子和正向散射拉曼信号从镜面钢表面被反射和回射,并聚焦到收集光学器件。不锈钢的空间分辨率至少可与CaF2相比,并且不受激光反射的影响。钢铁是CaF2成本的一小部分,光子的反射和聚焦提高了信噪比,可实现更快的映射。钢的低成本加上其拉曼信号增强的性能和坚固的耐用性表明,钢是生物学和临床RS的理想基材,因为它具有比常规使用的CaF2的关键优势。 ©2016作者。拉曼光谱学杂志,约翰·威利父子有限公司出版

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