Raman Spectroscopy of Biological Tissues

Abstract

We previously published a comprehensive review paper reviewing the Raman spectroscopy of biological molecules. This research area has expanded rapidly, which warranted an update to the existing review paper by adding the recently reported studies in literature. This article reviews some of the recent advances of Raman spectroscopy in relation to biomedical applications starting from natural tissues to cancer biology. Raman spectroscopy, an optical molecular detective, is a vibrational spectroscopic technique that has potential not only in cancer diagnosis but also in understanding progression of the disease. This article summarizes some of the most widely observed peak frequencies and their assignments. The aim of this review is to develop a database of molecular fingerprints, which will facilitate researchers in identifying the chemical structure of the biological tissues including most of the significant peaks reported both in the normal and cancerous tissues. It has covered a variety of Raman approaches and its quantitative and qualitative biochemical information. In addition, it covers the use of Raman spectroscopy to analyse a variety of different malignancies including breast, brain, cervical, gastrointestinal, lung, oral, and skin cancer. Multivariate analysis approaches used in these studies have also been covered.     

Raman spectroscopic analysis differentiates between breast cancer cell lines

Breast cancer incident rates are increasing in women worldwide with the highest incidence rates reported in developing countries. Major breast cancer screening approaches like mammography, ultrasound, clinical breast examination (CBE) and magnetic resonance imaging (MRI) are currently used but have their own limitations. Optical spectroscopy has attained great attention from biomedical researchers in recent years due to its non‐invasive and non‐destructive detection approach. Chemometrics is one of the powerful tools used in spectroscopic research to enhance its sensitivity. Raman spectroscopy, a vibrational spectroscopic approach, has been used to explore the chemical fingerprints of different biological tissues including normal and malignant types. This approach was used to characterize and differentiate two breast cancer and one normal breast cell lines (MDA‐MB‐436, MCF‐7 and MCF‐10A) using dispersive Raman spectroscopy. Raman spectra of the cell lines have revealed that basic differences in the concentration of biochemical compounds such as lipids, nucleic acids and protein Raman peaks were found to differ in intensity, and principal component analysis (PCA) was able to identify variations that lead to accurate and reliable separation of the three cell lines. Linear discriminant analysis (LDA) model of three cell lines was predicted with 100% sensitivity and 91% specificity. We have shown that a combination of Raman spectroscopy and chemometrics are capable of differentiation between breast cancer cell lines. These variations may be useful in identifying new spectral markers to differentiate different subtypes of breast cancer although this needs confirmation in a larger panel of cell lines as well as clinical material. Copyright © 2015 John Wiley & Sons, Ltd.     

Raman spectroscopic analysis of breast cancer tissues: identifying differences between normal,invasive ductal carcinoma and ductal carcinoma in situ of the breast tissue

A relatively non‐destructive method employing Raman spectroscopy for the analysis of histopathological specimens is described. Raman spectroscopy has allowed qualitative analysis of the same specimen used for histopathological evaluation. Breast cancer tissues have been analysed to demonstrate the feasibility of the chemical changes taking place in the biological tissue, which can be identified precisely, and the results are reproducible. Raman analysis of tissue sections provides distinct spectra that can be used to distinguish between the nuclear grades of ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) of the breast. Sixty cases of breast carcinoma including DCIS and IDC and seven cases of normal breast tissues were studied employing the Raman spectroscopic technique. This study reports for the first time spectral differences between DCIS grades. It is concluded that Raman spectroscopy can objectively distinguish between DCIS and IDC grades and is non‐destructive and reproducible. It should become possible in future to use Raman spectroscopy as an informative and quantitative method suitable for classification of grades and diagnosis of breast carcinoma. Copyright © 2007 John Wiley & Sons, Ltd.     

激光拉曼光谱分析氢同位素的应用

拉曼光谱作为一种物质结构和成分分析的测试手段而被广泛应用.介绍分析了激光拉曼光谱法用于氢同位素分析的可行性,并综述介绍了国内外研究人员利用激光拉曼光谱在氚参与的放射反应监测分析、氢同位素定性检测、定量分析方法研究等方面开展的工作.     

乳腺肿瘤周边组织的拉曼光谱研究

使用REMSHAW显微共焦拉曼光谱仪测量了40例手术切除乳腺肿瘤周边(肿块边约5 mm)组织的拉曼光谱.通过对浸润性导管癌、乳腺增生、纤维腺瘤等乳腺肿瘤周边组织的拉曼光谱分析,发现不同乳腺肿瘤周边组织的拉曼光谱有显著差异.这些差异在肿瘤诊断中,可作为乳腺癌和其他乳腺肿瘤拉曼光谱的特征,为乳腺疾病诊断提供依据.乳腺肿瘤周边组织的拉曼光谱对乳腺肿瘤诊断具有重要的价值.     

拉曼光谱在血红蛋白结构及功能研究中的应用进展

血红蛋白发挥多种重要生理功能,但对其结构及功能的认识尚不能满足疾病诊治的需求。拉曼光谱在血红蛋白结构及功能研究中具有很大应用潜力,不但可以检测血红蛋白中血红素及其周围分子结构变化,还可以反映血红蛋白反应动力学具体过程。同时,血红蛋白拉曼光谱在疾病状态下异常血红蛋白检测,血氧饱和度定量测定及血液代用品的高铁血红蛋白含量检测中突显优势。本文综述了拉曼光谱在血红蛋白结构及功能领域中的研究,简述了拉曼光谱在一些病变血红蛋白诊断中的研究进展,分析了影响血红蛋白拉曼光谱检测的因素,以促进拉曼光谱技术在血红蛋白结构和功能研究中的应用。     

拉曼光谱在天然纤维素结构研究中的应用进展

纤维素是木质纤维生物质细胞壁的骨架物质,也是生物燃料制备过程中重要的前驱体。作为重要的天然有机高分子,纤维素分子结构的研究备受关注。拉曼光谱仪因其较高的分辨率及无损检测的特点可在多尺度研究天然纤维素复杂分子链及聚集态结构。本文在比较了色散型拉曼光谱仪和傅里叶变换拉曼光谱仪的构造及相关参数的基础上,详细综述了拉曼光谱技术在植物细胞壁纤维素微区分布、天然纤维素酶解发酵、分子链空间取向、分子形变、结晶度与多晶态转变等方面的研究进展。并对拉曼光谱技术在天然纤维素分子结构研究中存在的问题进行了总结,提出了可能的解决方案,以促进拉曼光谱技术在天然有机高分子研究领域的应用。     

拉曼光谱技术在病原微生物检测中的应用

病原微生物是指可侵犯人体,引起感染的微生物,临床上由病原微生物感染引发的疾病极为常见。传统的临床病原菌诊断主要依赖于细菌培养,但此方法耗时长,往往需要2～5 d才能得到检测结果,并且存在部分细菌培养困难甚至无法培养的问题。在无法鉴别菌种以及药物敏感性的情况下医生凭借经验使用广谱抗生素,加速了细菌耐药性的产生。因此,病原微生物的高灵敏快速检测方法研究成为重要研究方向。拉曼光谱技术是一种对待测样品进行原位、非侵入性检测的技术,可在单细胞水平上提供微生物细胞中不同生物分子的指纹图谱信息,通过这些信息可以确定微生物的种类、生理特征和突变表型等,实现对微生物样品的快速检测。随着激光光谱学的快速发展以及临床需求的不断增加,促使了以拉曼光谱检测技术为核心的亚技术诞生（如：表面增强拉曼光谱技术、傅里叶变换拉曼光谱技术、激光共振拉曼光谱技术、共聚焦显微拉曼光谱技术、相干反斯托克斯拉曼光谱以及受激拉曼光谱等相关技术）,同时改善了以往拉曼光谱技术信号强度弱的不足,以实现对微生物高精度的快速检测分析。凭借着其具有对样本的状态没有限制以及能够检测物质成分微小变化的优势,近年来对拉曼光谱在病原微生物领域的研究日渐增多。对微生物检测的研究现状进行了调查和分析,围绕着拉曼光谱技术原理对其在微生物检测中的应用进行了具体阐述,其中主要对该技术在病原微生物鉴定以及药敏检测中的研究进展展开讨论,并就其与传统检测技术之间的差别和优势进行分析,展示了拉曼光谱技术作为病原微生物的快速检测新方法的前景。     

甲烷水合物拉曼光谱法研究进展

介绍了甲烷在气相、水合相中的拉曼光谱特征,从水合物生成热力学、生成动力学、分解动力学和分解机理几方面对甲烷水合物实验室拉曼光谱分析和深海拉曼光谱检测的最新进展进行了综述。生成热力学方面重点介绍了基于拉曼光谱技术的水合物生成条件的原位观测、水合物结构的鉴定及水合物孔穴占有率和水合数的求算,生成动力学方面主要介绍了水合物生成过程中孔穴形成随时间的变化关系及水合物形成后流体中甲烷浓度的变化规律等内容。水合物分解方面着重介绍了水合物分解的微观机理、孔穴占有率的变化规律及多孔介质中水合物分解速率表达式。针对目前拉曼光谱法研究水合物存在的问题,对未来的发展方向和重点提出了建议。     

共振拉曼光谱技术应用综述

拉曼光谱是提供物质结信息的强有力工具。但由于拉曼散射信号弱,灵敏度低,因此应用范围受到限制。而在共振拉曼光谱(RRS)中,由于激发光源频率落在分子的某一电子吸收带内,分子吸收光子向电子激发态的跃迁变成了共振吸收,因此对入射光的吸收强度大大增加。与常规拉曼光谱相比,RRS能够提高信号强度的106倍。因此,RRS检测技术以其更高的灵敏度和选择性而具有更广的应用,特别是在生物学及医学等领域。如：(1)生物基质中的类胡萝卜素和叶绿素等色素分析;(2)细胞、蛋白质和DNA等有机物研究以及一些临床疾病诊断。RRS可以得到在常规拉曼光谱中隐藏的、更为重要的分子结构信息。RRS总是在很低的浓度下测试,且共振拉曼增强的谱线是属于产生电子吸收的基团,这对于有色物和生物样品尤为重要。因为很多这类样品的活性部位接近于生色基团,且研究对象往往是生物大分子的某一部分,所以在研究生物物质的结构和功能的关系时,RRS起着重要作用。近年来,由于光谱技术的发展使得RRS检测技术得到创新与延伸,如液芯光纤共振拉曼光谱和透射共振拉曼光谱等新技术的应用。通过对近几年有关RRS技术应用的原始论文、数据和主要观点进行归纳整理与分析提炼,介绍了RRS这一专题的历史背景和研究现状,分别对共振拉曼光谱的色素检测、生物检测和爆炸物检测等应用领域展开详细的综述,并介绍了相关新技术的发展应用。随着光谱技术的快速发展,RRS必将在科研领域拥有其他光谱技术不可取代的重要地位。     

乙醇和甲醇混合溶液的拉曼光谱法研究

近年来激光技术的快速发展,使得拉曼光谱技术成为激光分析研究领域中的热门之一。拉曼光谱技术广泛地应用于结构化学分析。本文对甲醇和乙醇混合液的拉曼光谱进行了较详尽的研究分析,通过该实验可以有助于学生对激光拉曼技术的进一步了解。     

拉曼光谱在油气地质应用中的研究进展

拉曼光谱具有简单、快速、原位、微区、无损、高分辨率和高灵敏度等优点,可以分析物质的成分与分子结构信息,是开展地质研究的有力工具。通过回顾近年来拉曼光谱的研究进展,结合实际的油气地质研究工作及分析测试经验,对拉曼光谱在油气地质研究中的应用进行总结,并讨论现阶段应用过程中存在问题及未来的发展方向。拉曼光谱在油气地质中的应用主要分为三个方面：（1）矿物与流体包裹体分析,可以确定成岩-成藏流体类型及组成、成岩-成藏作用过程,包括岩矿鉴定、流体包裹体中流体体系分析、水-岩相互作用过程研究、地质样品同位素研究等;（2）不同类型有机质成熟度分析,可以用于恢复热史、油气成藏期次约束;（3）流体包裹体压力恢复,可以研究油气藏的形成与演化过程。主要方式为利用流体包裹体中气体的拉曼特征峰位移变化量、气体溶解度恢复流体包裹体内压及捕获压力。在实际油气地质研究中,多种因素制约拉曼光谱的定量/半定量应用,主要包括：地质样品复杂性与特殊性、样品处理方式、拉曼测试参数与测试环境、拉曼光谱数据处理与分析方式。拉曼光谱分析测试流程标准化、挑选和制备校准标样;拉曼光谱与冷热台、水热金刚石压腔（HDAC）、高压可视反应腔（HPOC）、扫描电镜（SEM）、电子探针（EPMA）等仪器联用,开展复杂体系原位、实时、不同温压条件下测试,是进一步将拉曼光谱应用到油气地质中的研究方向。     

鼻咽癌组织拉曼光谱研究进展

鼻咽癌是东南亚地区及中国南方地区高发的恶性肿瘤,具有独特的地理、种族分布特点,并且存在恶性程度高、预后差、早期诊断困难等问题。拉曼光谱技术是基于非弹性光散射基本原理的一种快速且无损的检测方法,能够在分子振动水平上提供生化成分等信息。综述基于拉曼光谱技术的鼻咽癌组织研究最新进展。主要介绍了国内外小组采用拉曼光谱及表面增强拉曼光谱(SERS)进行鼻咽癌组织检测研究的概况,其中重点介绍该研究小组近期在鼻咽癌组织的高波数拉曼光谱、鼻咽癌组织涂片的拉曼光谱,以及研发的人活体鼻咽癌组织内镜检测装置及其临床实验情况。最后,对鼻咽癌组织拉曼光谱研究的发展前景进行了展望。     

Recent advances in linear and nonlinear Raman spectroscopy. Part VII

The aim of this paper is to provide an overview of advances in the field of Raman spectroscopy as reflected in articles published each year in the Journal of Raman Spectroscopy as well as in trends across related journals publishing in this research area. The context for this review is derived from statistical data on article counts obtained from Thomson Reuters ISI Web of Knowledge by year and by subfield of Raman spectroscopy. Additional information is gleaned from presentations featuring Raman spectroscopy presented at the International Conference on Advanced Vibrational Spectroscopy in Kobe Japan in August 2013 and at SCIX 2013 sponsored by the Federation of Analytical Chemistry and Spectroscopy Societies in Milwaukee, Wisconsin, USA, October 2013. Papers published in the Journal of Raman Spectroscopy in 2012 are highlighted in this review and reflect topics and advances at the frontier of Raman spectroscopy, a field that is expanding rapidly as a sensitive photonic probe of matter at the molecular level in an ever widening sphere of novel applications. Copyright © 2013 John Wiley & Sons, Ltd.     

Applications of Raman Spectroscopy in Agricultural Products and Food Analysis: A Review

Abstract

Raman spectroscopy has been gaining popularity as an analytical tool due to advances in development of Raman spectrometry and the power of personal computers. Due to to its narrow and highly resolved bands, Raman spectroscopy allows for nondestructive extraction of chemical and physical information about samples and aids in rapid on-line analysis without any special sample preparation. In this review, Raman spectroscopic techniques such as dispersive Raman spectroscopy, Fourier transform Raman spectroscopy, surface-enhanced Raman spectroscopy, and spatially offset Raman spectroscopy are briefly introduced. In addition, applications of Raman spectroscopy are explored, within various fields of agricultural products and food, including fruits and vegetables, crops, meat and dairy products, oil, as well as beverages. In addition, some discussion on the importance of Raman spectroscopy as fundamental and applied research of agricultural products and food is provided.     

塑化剂邻苯二甲酸二辛酯的拉曼光谱计算及分析

近几年,国内有关塑化剂的食品安全事件频发,迫切需求加强对于食品中塑化剂的检测研究工作。邻苯二甲酸二辛酯是塑化剂的一种。理论计算和实验测量了邻苯二甲酸二辛酯的拉曼光谱。利用密度泛函理论DFT(B3LYP)的三参数混合方法对其进行了空间结构优化;然后分别用(Hartree-Fock)HF/3-21G和DFT/3-21G两种算法计算了它的拉曼光谱和红外光谱,给出了光谱强度图。实验测量了分析纯级邻苯二甲酸二辛酯的拉曼光谱,并和理论计算结果进行了对比,显示了很好的一致性,与此同时由于计算方法的不同,从图中可以看出Gaussian计算的HF和DFT算法无论是峰的位置还是相对强度等都均存在差别。还给出了常用塑化剂邻苯二甲酸二辛酯的各个键长、键角等空间结构参数,并对邻苯二甲酸二辛酯分子在400～3 500 cm-1区间的振动谱做了指认。报道了常用塑化剂邻苯二甲酸二辛酯的拉曼光谱研究工作。这些工作将有助于拉曼光谱技术在食品检测领域的研究和应用。     

拉曼光谱技术在农产品质量安全检测中的应用

农产品的质量安全与我们老百姓的身体健康和生命安全密不可分。传统的化学检测方法具有需要样品前处理,操作过程复杂以及破坏样品等诸多缺陷。拉曼光谱技术作为一种分析、测试物质分子结构强有力的表征手段,可以快速实现样品的无损伤、定性定量检测分析。随着拉曼光谱技术的不断完善和应用范围的逐渐拓宽,拉曼光谱技术在农产品的质量安全检测中发挥着极其重要作用,并且具有广阔的应用前景。目前,已经有大量的基于拉曼光谱技术检测农产品质量安全的相关研究报道,为了解拉曼光谱技术的检测原理以及发展现状,并跟踪国内外最新研究进展,简述了拉曼光谱技术的基本原理及其发展、拉曼光谱检测装置,深入综述了拉曼光谱技术在果蔬、禽畜、粮食质量安全检测中的最新研究进展,指出了拉曼光谱技术应用在农产品质量安全检测中的现存的技术问题。另外,还简要介绍了国内外部分拉曼光谱仪的部分信息和便携式拉曼光谱仪专利申请状况,展望了该项技术的研究方向和应用前景。     

Application of Raman Spectroscopy Technique to Agricultural Products Quality and Safety DeterminationSCIEI北大核心CSCD

The quality and safety of agricultural products and people health are inseparable. Using the conventional chemical methods which have so many defects, such as sample pretreatment, complicated operation process and destroying the samples. Raman spectroscopy as a powerful tool of analysing and testing molecular structure, can implement samples quickly without damage, qualitative and quantitative detection analysis. With the continuous improvement and the scope of the application of Raman spectroscopy technology gradually widen, Raman spectroscopy technique plays an important role in agricultural products quality and safety determination, and has wide application prospects. There have been a lot of related research reports based on Raman spectroscopy detection on agricultural product quality safety at present. For the understanding of the principle of detection and the current development situation of Raman spectroscopy, as well as tracking the latest research progress both at home and abroad, the basic principles and the development of Raman spectroscopy as well as the detection device were introduced briefly. The latest research progress of quality and safety determination in fruits and vegetables, livestock and grain by Raman spectroscopy technique were reviewed deeply. Its technical problems for agricultural products quality and safety determination were pointed out. In addition, the text also briefly introduces some information of Raman spectrometer and the application for patent of the portable Raman spectrometer, prospects the future research and application.     

应用拉曼光谱分析变压器油中溶解乙酸含量

乙酸是变压器油纸绝缘老化所生成酸类物质的主要成分;分析变压器油中溶解乙酸含量对准确评估运行变压器的老化状态具有重要意义。拉曼光谱技术是基于拉曼效应的一种分子分析技术,能很好地用于物质的非接触式原位检测。论文开展了变压器油中溶解乙酸含量拉曼光谱检测方法研究： 利用Gaussian 09W软件分析了乙酸分子的拉曼振动特性,对实测乙酸拉曼谱峰的振动模式进行了指认;基于实验室搭建的激光拉曼光谱液体检测平台,对不同乙酸含量的变压器油样进行了原位检测;选定891 cm-1作为变压器油中溶解乙酸分子的拉曼特征峰,基于乙酸891 cm-1与变压器油932 cm-1特征峰面积比值和最小二乘法建立了乙酸的定量分析方法,检测限可达0.08 mg·mL-1。实验结果表明： 激光拉曼光谱可应用于油中溶解乙酸含量原位检测并具有良好的检测稳定性和重复性,为变压器油中溶解乙酸含量的快速、无损检测提供了一种新方法。     

拉曼光谱技术在病理诊断中的研究进展

拉曼光谱技术能够提供与物质特定分子结构相关的光谱信息,可用于识别生物组织微小的生化变异,具有快速、实时、无损、无需样本预处理等优点,在临床病理诊断领域极具应用前景。与常规组织病理学分析相比,拉曼光谱技术能够直接检测活体组织,简化了分析程序,缩短了诊断时间。人体病变组织的细胞分子组成和结构可能发生变化,这为拉曼光谱技术在组织病理诊断中的应用提供了检测依据。基于组织分子组成与结构的差异,结合机器学习和化学计量学方法,拉曼光谱技术可以提供客观的诊断信息,实现快速、低侵入的病理诊断。回顾了近十年来拉曼光谱技术在组织病理诊断中的研究进展,对取得的关键成果进行了总结,阐述了当前离体和活体应用拉曼光谱技术的一些关键问题。针对离体拉曼光谱检测,重点评估福尔马林固定石蜡包埋样本、冷冻样本和新鲜组织样本等离体样本的适用情况;阐述拉曼光谱数据收集的关键技术,包括适用光源、光谱范围,以及病理样本光谱采集的方式等。对于活体拉曼光谱检测,重点介绍了活体检测研究中拉曼光谱技术应用的两种形式：结合医用内窥镜进行体内检测,以及开放手术中的直接检测;综述了临床适用的拉曼系统,重点介绍了当前活体拉曼研究中应用的光纤探头。同时,文章也讨论了拉曼光谱数据的处理与分析方法,通过光谱预处理,特征提取与分类识别,构建拉曼光谱病理诊断模型,在小样本范围能够获得较好的诊断结果。考虑临床实际应用,仍需要不断优化分析方法,实现拉曼光谱与生化信息的关联,将样本个体差异的影响纳入分类模型中,以提升模型性能。文章对拉曼光谱应用于病理诊断中的关键问题进行了讨论,为进一步开展研究提供参考。未来需要更深入和广泛地开展离体和活体研究,以促进拉曼光谱技术在临床中的应用。