Raman and Infra‐Red Microspectroscopy: Towards Quantitative Evaluation for Clinical Research by Ratiometric Analysis
Raman and Infra‐Red Microspectroscopy: Towards Quantitative Evaluation for Clinical Research by Ratiometric Analysis
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DOI：
10.1002/chin.201621278
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Srividya Kumar;Taru Verma;Ria Mukherjee;F. Ariese;K. Somasundaram;S. Umapathy
中科院分区：
文献类型：
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作者：
Srividya Kumar;Taru Verma;Ria Mukherjee;F. Ariese;K. Somasundaram;S. Umapathy
生物分子结构解析是研究生命基本过程的主要技术之一。由于疾病等各种原因，这些过程被调节、阻碍或改变，这就是为什么生物分子分析和成像在诊断、治疗预后和监测中发挥着重要作用。振动光谱(红外光谱和拉曼光谱)是一种分子键特异性技术，可以帮助研究人员解释化学结构。基于与显微镜的结合，振动显微光谱学目前正在成为生物医学研究的重要工具，具有细胞和亚细胞水平的空间分辨率。这些技术提供了各种优势，使自然状态下的无标记生物分子指纹图谱成为可能。然而，从光谱中破译所需信息的复杂性阻碍了它们进入临床。如今，随着自动化算法的出现，振动显微光谱学在光谱病理学领域表现出色。然而，研究人员应该意识到，基于绝对波段强度的量化可能会受到仪器参数、样品厚度、水分含量、底物背景和其他可能的伪影的影响。在这篇综述中，我们将详细讨论这些实际问题及其对生物分子定量的影响。在许多情况下，比率分析可以帮助绕过这些问题，并使生物样本的定量研究成为可能，包括一维、二维和三维的比率成像。我们提供了一个广泛的概述，从最近的科学文献中的红外和拉曼波段比率用于研究生物系统和疾病诊断和治疗预后。
Biomolecular structure elucidation is one of the major techniques for studying the basic processes of life. These processes get modulated, hindered or altered due to various causes like diseases, which is why biomolecular analysis and imaging play an important role in diagnosis, treatment prognosis and monitoring. Vibrational spectroscopy (IR and Raman), which is a molecular bond specific technique, can assist the researcher in chemical structure interpretation. Based on the combination with microscopy, vibrational microspectroscopy is currently emerging as an important tool for biomedical research, with a spatial resolution at the cellular and sub-cellular level. These techniques offer various advantages, enabling label-free, biomolecular fingerprinting in the native state. However, the complexity involved in deciphering the required information from a spectrum hampered their entry into the clinic. Today with the advent of automated algorithms, vibrational microspectroscopy excels in the field of spectropathology. However, researchers should be aware of how quantification based on absolute band intensities may be affected by instrumental parameters, sample thickness, water content, substrate backgrounds and other possible artefacts. In this review these practical issues and their effects on the quantification of biomolecules will be discussed in detail. In many cases ratiometric analysis can help to circumvent these problems and enable the quantitative study of biological samples, including ratiometric imaging in 1D, 2D and 3D. We provide an extensive overview from the recent scientific literature on IR and Raman band ratios used for studying biological systems and for disease diagnosis and treatment prognosis.