拉曼光谱分析技术在细胞生物学研究中的应用进展

细胞是生物体结构和功能的基本单位,自被发现以来新的研究方法不断涌现。单细胞拉曼光谱能提供细胞内核酸、蛋白质、脂质含量等大量信息,可在不损伤细胞的条件下实时动态地监测细胞分子结构变化,亦可获得细胞的“分子指纹”,具有敏感性高、实时检测、活样品不需固定或染色、不损伤细胞等众多特点。近年来国内外研究者将拉曼光谱应用于细胞药物处理、细胞水平疾病诊断、单细胞生命活动监测、亚细胞结构等研究,取得了不同程度的进展。随着研究的深入,拉曼光谱分析技术必将在干细胞,癌症研究、细胞分选、药物筛选等领域大有作为。     

唇腺炎性病变组织的共聚焦显微拉曼光谱特征研究

目的:研究唇腺炎性病变组织的拉曼光谱指纹特征,为拉曼光谱技术临床鉴别诊断唇腺炎性病变提供理论基础。方法:收集舍格伦综合征病变唇腺30例、唇腺急性炎症组织18例及正常唇腺组织30例,应用激光共聚焦显微拉曼光谱仪对唇腺组织进行拉曼光谱检测。应用主成分分析法(Principal component analysis,PCA)及判别函数(Discrimination function analysis,DFA)对光谱数据进行分析,研究唇腺组织光谱指纹诊断价值。结果:唇腺炎症组织与正常组织光谱间存在光谱指纹差异,这些差异代表了某些蛋白、核酸及脂类物质等生物大分子发生改变。PCA-DFA分析发现这些差异性拉曼光谱具有鉴别诊断价值,可以区分不同唇腺组织,总体诊断准确率达91.8%,经交互验证后准确率为89.4%。结论:不同唇腺炎症组织及正常组织间拉曼光谱存在差异,不仅揭示生物大分子改变,还具有临床鉴别诊断价值。拉曼光谱技术在唇腺炎性病变组织鉴别诊断具有巨大应用潜力。     

非标记表面增强拉曼光谱在DNA检测中的应用

表面增强拉曼光谱(SERS)是一种超灵敏的生化分析技术,已经被广泛运用于细胞、核酸、蛋白质等生物分子的检测,在生物医学领域表现出了巨大的应用潜力。近年来,将表面增强拉曼光谱技术应用于遗传物质DNA的精准检测,引起了人们广泛的关注。本文简要叙述了表面增强拉曼光谱技术的基本原理及其在DNA检测中的优势,主要介绍了非标记的DNA-SERS检测应用进展,其中包括本项目组的相关工作。研究表明,非标记DNA-SERS技术有望成为一种快速、准确的临床诊断方式。     

基于光镊与拉曼光谱的红菇担孢子分析

拉曼光谱作为分子振动光谱,具有无损、无需试剂染色和快速方便的特点,在生物学领域的应用越来越广泛。拉曼镊子是光镊与拉曼光谱的结合,应用拉曼镊子技术收集单个红菇担孢子的拉曼光谱,分析单个担孢子的主要成分,不同的储存方法对担孢子成分的影响等。结果发现,单个担孢子的拉曼光谱反映了担孢子内含物的基本组成,红菇担孢子的孢内主要成分是脂类物质;不同种类的红菇担孢子的拉曼光谱基本相同,经过多年保存的孢子仍然保持其孢子内的主要成分不变。各随机选择10个担孢子的拉曼光谱进行主成分(PCA)判别分析,结果显示红菇属内不同种类的孢子无法区分。结果表明单孢子拉曼光谱可以应用于分析孢子内成分的变化,但应用于真菌分类的可能性有待进一步探索。     

拉曼光谱检测微生物的研究方法和进展北大核心

快速准确地识别和鉴定微生物对于环境科、食品质量以及医学诊断等领域研究至关重要。拉曼光谱(Raman spectroscopy)已经被证明是一种能够实现微生物快速诊断的新技术,在提供微生物指纹图谱信息的同时,能够快速、非标记、无创、敏感地在固体和液体环境中实现微生物单细胞水平的检测。本文简单介绍了拉曼光谱的基本概念和原理,重点综述了拉曼光谱微生物检测应用中的样品处理方法及光谱数据处理方法。除此之外,本文概括了拉曼光谱在细菌、病毒和真菌中的应用,其中单独概括了拉曼在细菌快速鉴定和抗生素药敏检测中的应用。最后,本文阐述了拉曼光谱在微生物检测中的挑战和展望。     

基于单细菌共焦拉曼光谱的细菌快速检测

【背景】目前利用共焦拉曼光谱技术进行成像和成分鉴别方面的研究较多,但如何快速检测与鉴别多种细菌方面的研究较少。【目的】基于共焦拉曼光谱技术,建立一种在单细菌水平上实现病原微生物快速分类鉴定的方法。【方法】以大肠杆菌为研究对象,利用共焦拉曼光谱技术在单细菌水平上进行了激发波长的优化试验,并研究了大肠杆菌存放时间对单细菌拉曼光谱信息的影响。同时,对白色葡萄球菌、大肠杆菌、金黄色葡萄球菌、沙门氏菌和铜绿假单胞菌进行了共焦拉曼光谱测试,并对5种细菌进行单细菌拉曼光谱的归属分析,设计共焦拉曼光谱技术结合支持向量机(support vector machine,SVM)模型学习算法,进行了5种细菌的快速分类鉴别。【结果】对于单细菌拉曼光谱探测,532、633和785 nm这3种常见的拉曼探测波长中,532 nm具有更好的激发效率和光谱信噪比。结合SVM模型对5种细菌的识别分类,SVM模型的灵敏度和特异性达到了96.00%以上,整体准确率为98.25%。不同存放时间下大肠杆菌拉曼光谱的重复性和稳定性都很好,且SVM模型匹配率均在90.00%以上。【结论】单细菌拉曼光谱结合SVM模型可对5种细菌进行快...     

功能红细胞研究的拉曼光谱技术与进展

红细胞在人体新陈代谢和物质运输方面起着非常重要的作用。作为一种无损、快速而灵敏的光谱学检测方法,拉曼光谱技术在生物医学领域的应用日渐广泛。本文综述了近年来拉曼光谱技术在红细胞及血红蛋白研究中的应用进展,详尽评述当前研究功能红细胞中较有优势的拉曼技术—显微拉曼光谱技术和光镊拉曼光谱技术,并对其技术的改进做出总结,以期为利用激光拉曼光谱研究红细胞提供较系统的参考。     

卵巢癌患者血清的拉曼光谱研究初探

目的:探究拉曼光谱技术应用于卵巢癌研究的可能性。方法:收集卵巢癌患者血清及健康人血清各20例,用激光共聚焦显微拉曼光谱仪进行检测。结果:两组血清的平均拉曼光谱形态和谱峰基本相似,但在约1010、1158、1283、1520、1646、2307和2661cm-17个拉曼频移附近,卵巢癌患者血清的拉曼光谱谱峰强度明显低于健康对照组,而在其余大部分波段,卵巢癌患者血清的拉曼光谱强度均高于健康对照组。结论:拉曼光谱技术可以初步区分卵巢癌及健康人血清,值得进一步研究和探讨其临床应用价值。     

拉曼光谱技术在微生物学中的应用

拉曼光谱具有快速、灵敏、无损、实时监测等显著特点,在微生物学领域得到广泛应用。分别介绍共焦显微拉曼光谱、共振拉曼光谱、表面增强拉曼光谱、拉曼成像、相干反斯托克斯拉曼光谱、激光镊子拉曼光谱和Raman-FISH的原理和特点,并重点总结和分析不同拉曼光谱技术在微生物的结构、化学组成,以及代谢过程等相关研究中的应用优势。合理利用这些技术在基础微生物、发酵微生物和微生物诊断等方面具有潜在的应用价值。     

鼻咽癌细胞组织与血液的SERS光谱研究进展

基于表面增强拉曼光谱(SERS)技术对于人体细胞组织与血液的检测和研究,SERS光谱技术能够发现正常组织与病变组织的差异性,为医学临床上实现癌症的早期诊断提供了科学依据。由于鼻咽癌不具有明显的病变特征、病灶的位置难以通过常规医学手段检测。因此利用SERS光谱技术,应用于鼻咽癌细胞组织与血液的研究,可提高鼻咽癌患者的生存率。对鼻咽癌细胞组织与血液SERS光谱分析和诊断的探索研究,有助于SERS光谱技术发展成为一种在生物医学领域中的分析检测手段,在医学临床诊断上具有潜在的应用前景。     

拉曼光谱技术在肿瘤检测中的应用

癌症是威胁人类健康和生命的严重疾病之一,早期诊断与及时治疗是提高癌症患者生存率的最有效途径。激光拉曼光谱技术作为一种非侵入性的检测技术,可以无损伤地提供丰富的分子结构特征和物质成分信息,从分子水平上反映癌变组织与正常组织之间的结构差异,从而可用于癌症的早期诊断。综述了激光拉曼光谱技术在皮肤癌、鼻咽癌、肺癌、胃癌、结肠癌、乳腺癌及前列腺癌诊断中的研究进展,并对拉曼光谱技术在癌症诊断中的发展方向和应用前景作了进一步的展望,为癌症的早期检测和诊断技术的应用研究提供参考依据。     

Research Progress of Raman Spectroscopy in Drug Analysis

Raman spectroscopy is a spectroscopic analysis technique that enables rapid qualitative and quantitative detection based on inelastic collision and Raman scattering intensity. This review detailed the generation principle, instrument composition, influencing factors, and common classifications of Raman spectrum. Furthermore, it summarized and forecast the research progress of Raman spectroscopy in the field of drug analysis simultaneously over the past decade, including the identification of active pharmaceutical ingredients (APIs), qualitative and quantitative studies of pharmaceutical preparations, detection of illicit drugs, the identification of Chinese herbal medicines, and the combination with other technologies. The development of Raman spectroscopy in other fields is additionally summarized.     

二甲基亚砜对离体猪皮肤组织的拉曼光谱的影响

拉曼光谱技术作为鉴定生物分子种类最有力的分析工具之一,具有快速、简单、无损、准确等优点.目前,拉曼光谱已被国内外学者广泛开展了在人体组织的应用研究,但由于生物组织具有高散射性,因此限制了拉曼光谱对其的检测深度.本文主要采用光透明剂--二甲基亚砜对组织拉曼光谱的影响进行研究,对离体猪皮组织的不同深度（100 μm,200 μm,300 μm,400 μm）拉曼光谱强度随处理前、后不同时间（0 min,10 min,20 min,30 min,60 min）的变化进行对比.结果发现,不管是经二甲基亚砜处理前还是后,都出现随着距猪皮组织表面的深度加深,其拉曼光谱强度都不断减少;同时发现各层猪皮组织随着处理后时间的加长,其特征峰的强度不断加强,信噪比逐渐提高,在处理后的60 min效果最好;而且出现了在经二甲亚砜处理前看不见的峰（1 126 cm-1和1 426 cm-1）.结果表明：5%DMSO对组织的处理能增强其拉曼光谱的强度,同时也能使拉曼光谱仪的信噪比提高,并且使组织谱图中特征峰也得到相应的增加.     

拉曼光谱技术在病原菌感染诊断中的应用进展

快速准确诊断感染性疾病病原体是遏制超级细菌传播和抗生素滥用的重要防线。目前,临床病原菌感染诊断十分依赖于培养手段,导致检测周期长达数日,不但影响了患者的及时诊治,还间接导致抗生素的滥用。拉曼光谱技术是一种无损、高灵敏的分子指纹图谱检测技术,近年来在生物学领域得到广泛应用,其具有的免培养、快速、高特异性、低成本等优点为病原菌感染的诊断提供了新方案。本文阐述了拉曼光谱技术的原理和特点,综述了其在病原菌鉴定和抗菌药物敏感性试验方面的应用进展。     

Raman spectroscopic analysis of skin as a diagnostic tool for Human African Trypanosomiasis

Human African Trypanosomiasis (HAT) has been responsible for several deadly epidemics throughout the 20th century, but a renewed commitment to disease control has significantly reduced new cases and motivated a target for the elimination of Trypanosoma brucei gambiense-HAT by 2030. However, the recent identification of latent human infections, and the detection of trypanosomes in extravascular tissues hidden from current diagnostic tools, such as the skin, has added new complexity to identifying infected individuals. New and improved diagnostic tests to detect Trypanosoma brucei infection by interrogating the skin are therefore needed. Recent advances have improved the cost, sensitivity and portability of Raman spectroscopy technology for non-invasive medical diagnostics, making it an attractive tool for gambiense-HAT detection. The aim of this work was to assess and develop a new non-invasive diagnostic method for T. brucei through Raman spectroscopy of the skin. Infections were performed in an established murine disease model using the animal-infective Trypanosoma brucei brucei subspecies. The skin of infected and matched control mice was scrutinized ex vivo using a confocal Raman microscope with 532 nm excitation and in situ at 785 nm excitation with a portable field-compatible instrument. Spectral evaluation and Principal Component Analysis confirmed discrimination of T. brucei-infected from uninfected tissue, and a characterisation of biochemical changes in lipids and proteins in parasite-infected skin indicated by prominent Raman peak intensities was performed. This study is the first to demonstrate the application of Raman spectroscopy for the detection of T. brucei by targeting the skin of the host. The technique has significant potential to discriminate between infected and non-infected tissue and could represent a unique, non-invasive diagnostic tool in the goal for elimination of gambiense-HAT as well as for Animal African Trypanosomiasis (AAT).     

激光拉曼光谱技术在生物分子DNA研究中的应用和进展

激光技术的兴起使拉曼光谱成为激光分析中最活跃的研究领域之一,已被广泛地用于物质成分的分析和分子结构的鉴定。本文综述了拉曼技术在DNA研究中近年来的最新进展,包括：DNA的常规拉曼光谱分析;DNA的激光共振拉曼光谱分析;DNA在金属表面或电极上吸附行为的表面增强拉曼光谱研究;DNA的傅立叶变换拉曼光谱研究等。并对拉曼光谱技术在DNA等生物大分子领域中的研究前景做了进一步的展望。     

Detection of Vero Cells Infected with Herpes Simplex Types 1 and 2 and Varicella Zoster Viruses Using Raman Spectroscopy and Advanced Statistical Methods

Of the eight members of the herpes family of viruses, HSV1, HSV2, and varicella zoster are the most common and are mainly involved in cutaneous disorders. These viruses usually are not life-threatening, but in some cases they might cause serious infections to the eyes and the brain that can lead to blindness and possibly death. An effective drug (acyclovir and its derivatives) is available against these viruses. Therefore, early detection and identification of these viral infections is highly important for an effective treatment. Raman spectroscopy, which has been widely used in the past years in medicine and biology, was used as a powerful spectroscopic tool for the detection and identification of these viral infections in cell culture, due to its sensitivity, rapidity and reliability. Our results showed that it was possible to differentiate, with a 97% identification success rate, the uninfected Vero cells that served as a control, from the Vero cells that were infected with HSV-1, HSV-2, and VZV. For that, linear discriminant analysis (LDA) was performed on the Raman spectra after principal component analysis (PCA) with a leave one out (LOO) approach. Raman spectroscopy in tandem with PCA and LDA enable to differentiate among the different herpes viral infections of Vero cells in time span of few minutes with high accuracy rate. Understanding cell molecular changes due to herpes viral infections using Raman spectroscopy may help in early detection and effective treatment.     

The use of optical spectroscopy in combinatorial chemistry.

Infrared and Raman spectroscopy allow direct spectral analysis of the solid-phase, thus avoiding the tedious cleavage of compounds from the solid support. With diagnostic bands in starting materials or products, infrared and Raman spectroscopy are efficient in monitoring each reaction step directly on the solid phase. Consequently, infrared and Raman spectroscopy have evolved as the premier analytical methodology for direct analysis on the solid support. While infrared transmission spectroscopy is a general analytical method for resin samples, internal reflection spectroscopy is especially suited for solid polymer substrates known as "pins" or "crowns." Single bead analysis is done best by infrared microspectroscopy, whereas photoacoustic spectroscopy allows totally nondestructive analysis of resin samples. With an automated accessory, diffuse reflection spectroscopy provides a method for high throughput on-bead monitoring of solid-phase reactions. Providing identification based on molecular structure, HPLC-FTIR is, therefore, complementary to LC-MS. Additionally, Raman spectroscopy as a complement to infrared spectroscopy can be applied to resin samples and-using a Raman microscope-to single beads. Fluorometry as an extremely sensitive spectroscopic detection method allows rapid quantification of organic reactions directly on the resin.