基于芯片实验室技术的表面增强光谱学在分析科学中的应用

集成具有一序列微流控操作单元的芯片实验室技术,在微流控通道内铺陈金属纳米粒子(尤其是金、银以及铜纳米粒子)作为衬底,泵入多通道微纳升分析物,用于联用表面增强光谱在痕量、实时、原位、过程反应等检测中具有重要的意义。这种联用检测技术集成了芯片实验室和表面光谱两种技术的优点：芯片实验室技术集成流程式分步操作,实现筛选取样,分段、实时反应检测,减小样品量,稳定测试环境等优势以及表面增强光谱的光谱响应快,灵敏性和选择性强、原位检测等优点。借助于Drude模型以及适当的边界条件,外电场引发金属颗粒价电子的局域等离子振荡,并推导了产生共振的局域表面等离子增强以及受激感应偶极子振荡产生表面拉曼增强的物理电磁增强机制。综述了芯片实验室表面局域等离子检测在生物、医药、食品安全等方面的应用,检测通道的增加促使检测效率有较大的提高,同时检测限能力获得较大的突破。综述了芯片实验室技术结合表面增强拉曼光谱公共安全、生物医学、电化学和生物传感器等领域的应用, 表面增强拉曼光谱的高度灵敏性以及指纹性应用于痕量检测。根据芯片实验室技术在研究开发和应用已经获得不断的进展,结合3D打印技术,精准控制多通道结构尺寸,更好地满足设计的需求。表面等离子增强光谱以及表面增强拉曼光谱等表面光谱检测技术在应用上日趋成熟,获得突破传统显微镜的光学极限的分辨能力。这种联用技术在实际定性或者半定量痕量分析检测应用中具有光明的前景。     

基于表面增强拉曼技术的致病菌及其耐药性检测研究进展

随着抗菌药物广泛应用于临床,细菌耐药日益严重。实现快速、高灵敏、准确的细菌及其药物敏感性检测是缓解细菌耐药的关键环节。表面增强拉曼光谱(SERS)具有快速、灵敏、无损等优点,可直接获取分子指纹信息,它已成为一种有效的细菌及其耐药性检测技术。不同种类细菌的分子组成和结构存在差异、抗生素处理前后细菌的特征拉曼信号会发生变化,这为表面增强拉曼光谱技术在致病菌及其耐药性检测中的应用提供了依据。基于分子组成与结构的差异, 结合传统多分类数据分析以及机器学习算法,表面增强拉曼光谱技术可以提供客观的诊断信息。这篇综述回顾了近年来表面增强拉曼光谱技术对于致病菌及其耐药性检测的研究进展, 阐述了当前表面增强拉曼光谱技术应用于致病菌检测面临的问题。首先,讨论了致病菌及其耐药性检测中常用SERS基底的材料和结构：金纳米粒子、银纳米粒子、银包金纳米粒子以及新型纳米材料与纳米粒子结合形成的复合SERS基底。然后,概述了SERS检测中捕获细菌的方法,主要介绍了基于核酸适配体、免疫磁性分离、微流控系统以及静电结合的捕获方法,包括上述捕获方法的原理以及捕获方式,综述了以上捕获方法的研究进展。最后,总结了致病菌SERS光谱的各种数据分析方法,通过光谱预处理,特征提取与分类识别,以及构建致病菌SERS光谱诊断模型,实现致病菌及其耐药性的检测;比较了传统的数据分析方法以及机器学习分析方法,重点介绍了深度学习算法在致病菌及其耐药性SERS检测中的优势与应用。文章也对表面增强拉曼光谱应用于致病菌及其耐药性检测的关键问题进行了讨论,并对基于表面增强拉曼技术的致病菌及其耐药性检测方法进行了展望,以促进表面增强拉曼光谱技术在临床检测中的应用。     

荧光/表面增强拉曼散射双模式纳米光学探针的构建及性能研究

提出一种新型的荧光及表面增强拉曼散射（SERS）双模式光学纳米探针。首先,通过再沉淀-包覆法合成二氧化硅包覆香豆素6的纳米颗粒,再在二氧化硅表面静电吸附多聚赖氨酸分子形成包覆层,随后通过原位还原的方法在多聚赖氨酸壳层复合银纳米颗粒,最后在银纳米颗粒表面吸附拉曼分子即形成双模式纳米探针。该探针通过二氧化硅包覆的荧光分子产生荧光信号,以多聚赖氨酸表面的银纳米颗粒作为SERS增强基底,利用拉曼分子获得SERS信号,实现了荧光及SERS双模式成像。荧光与表面增强拉曼散射相结合的双模式分析技术可同时发挥二者的优点,提高成像的分辨率和灵敏度,在生物医学领域具有重要的应用价值。     

微流控SERS芯片及其生物传感应用

由于微流控芯片具有优异的集成性和灵活的可操作性,基于芯片上的检测方法被大量开发,发展十分迅速。其中,表面增强拉曼光谱（SERS）凭借其超高的灵敏度、独一无二的指纹谱和窄峰宽等特点成为一种广泛采用的检测手段。SERS微流控芯片集SERS检测技术与微流控芯片的优势于一体,一方面为SERS检测方法的重复性和可靠性提供了一个高效平台,另一方面推动了微流控芯片的功能拓展,在生物分子探测、细胞捕获乃至组织模拟等领域具有广阔的应用前景。本文在简要介绍SERS的原理及其生物传感应用的基础上,重点概述了SERS微流控芯片的构建及其在生物传感及检测中的应用,最后探讨了该研究方向存在的问题及发展方向。     

自组装银纳米粒子及其SERS增强效应

采用柠檬酸三钠还原硝酸银方法制备出银纳米粒子, 并通过在玻璃表面修饰3-氨基丙基-三乙氧基硅烷( APTES)对银纳米粒子进行自组装。利用紫外-可见(UV-Vis)吸收光谱和扫描电子显微镜(SEM)测试手段对样品进行分析和表征。由测试结果可知银纳米粒子的尺寸比较均匀, 组装致密度较高, 基本以亚单层的形式分布于基底表面。进一步研究了以结晶紫(CV)为探针分子的自组装基底的表面增强拉曼光谱(SERS), 计算发现该基底的拉曼增强因子数量级达106。结果表明: 银纳米粒子自组装基底具有良好的SERS增强效应, 为痕量CV的检测提供了有效的方法。     

肿节风的表面增强拉曼光谱检测

提出一种基于表面增强拉曼光谱的中药材肿节风饮片的检测方法。采用柠檬酸三钠还原硝酸银制备银溶胶,以银胶纳米粒子为增强基底测得肿节风茎切片的表面增强拉曼光谱(SERS)。发现银胶直接作用于药材表面的SERS信号明显增强,肿节风茎切片SERS光谱中在637,1 176,1 309,1 476,1 612 cm^-1处都可观察到明显的拉曼特征峰。通过一阶导数拉曼光谱分析技术和对照品异嗪皮啶谱峰指认,可将获得的SERS峰位分别归属于吡喃酮环、甲氧基和酚羟基分子结构。研究结果表明,SERS技术可为肿节风和其他中草药的生产和质量监控提供一种快速、方便和直接的检测方法。     

新型冠状病毒S蛋白在金纳米粒子中的表面增强拉曼效应

表面增强拉曼光谱技术因其高灵敏度、操作简单、快速检测等优点,被广泛用于病毒检测方面。国内外的病毒拉曼检测研究主要集中在检测病毒核酸以及组成核酸的各种碱基的表面增强拉曼光谱(SERS),但少见对病毒蛋白的SERS检测。以新型冠状病毒（SARS-CoV-2）的S蛋白为检测对象,采用无标记SERS检测方法,对比SARS-CoV-2固态、饱和液态S蛋白的普通拉曼光谱和选用40 nm金纳米粒子为基底的SARS-CoV-2低浓度S蛋白SERS光谱。结果表明,以40 nm金纳米粒子为基底,采用SERS技术检测SARS-CoV-2的S蛋白是完全可行的。SARS-CoV-2的S蛋白分子中的羧基与金纳米粒子发生了分子增强,氨基与金纳米粒子发生了电磁增强,从而使得SARS-CoV-2的S蛋白拉曼效应得到了增强,并使得峰位发生一定移动。实验获得了较好的SARS-CoV-2低浓度S蛋白SERS光谱,为建立敏感、特异、快速的SARS-CoV-2检测新技术提供了一种方法。     

用于SERS基底的Au-Pd核壳纳米颗粒和银纳米立方颗粒的制备及表征(英文)

本文研究了尺寸可控的Au -Pd核壳纳米粒子和银纳米立方颗粒的表面增强拉曼散射(SERS)活性。发现Au-Pd核壳纳米粒子的增强能力要比粗糙的钯电极强;银纳米立方颗粒的增强能力和粗糙的银电极相当。更为重要的是,银纳米立方颗粒既具有原子级平整的小单晶面又处于纳米尺度,因而它们可以作为粗糙表面和结构确定的单晶表面之间的桥梁,对其SERS效应的研究可以加深人们对SERS机理的认识。     

银纳米有序阵列SERS活性基底的制备及在膀胱肿瘤细胞检测中的应用

以多孔阳极氧化铝(porous anodic alumina,PAA)膜为模板,采用真空电子束蒸镀技术,分别在PAA多孔层以及阻挡层表面形成了银纳米孔和银纳米帽有序阵列表面增强拉曼散射(surface-enhancedRaman scattering,SERS)活性基底,并以膀胱肿瘤细胞作为分子探针,测试和分析了这两种SERS活性基底的表面增强拉曼光谱的特性。结果表明,两种SERS活性基底对膀胱肿瘤细胞的拉曼散射信号均有很好的增强作用。银纳米帽有序阵列SERS活性基底不仅具有较高的SERS增强和荧光猝灭效应,而且不存在与PAA膜中草酸根杂质相关的干扰峰,可获得膀胱肿瘤细胞拉曼散射光谱的更多细节信息。     

纳米银组装结构上罗丹明B的表面增强荧光效应

采用表面自组装技术,在玻璃表面构筑银纳米粒子的二维组装结构。银纳米粒子组装结构的表面等离子共振光谱中偶极子表面等离子体共振对组装结构更为敏感而表现出较大位移。组装银纳米粒子可极大增强罗丹明B的荧光。荧光的表面增强效应主要来自银纳米粒子对荧光分子所处区域的局部电磁场增强,银纳米粒子的表面分子修饰对其表面增强效应有较大的影响。     

Optical properties of Ag@cicada wing substrate deposited by Ag nanoparticles

In this paper, Ag nanoparticles were deposited on Ag@cicada wing array by using the cicada wings as templates to study its optical properties, including surface enhanced Raman scattering (SERS), polarization and surface enhanced fluorescence (SEF). The nanogaps between adjacent conical protrusion can be well dominated by adjusting the sputtering time and the optimal substrate AgNPs@Ag@cicada wing arrays have a noteworthy enhancement of SERS signal. Characterization of the prepared optimal substrate certified that it possesses the excellent SERS performances. Basically consistent SERS signal strength at the different polarization angles of the optimal substrate indicates that its polarization-independence. The SEF spectra shows that the optimal substrate has a slightly lower and unstable enhancement at this initial stage of repeated examination due to the weak adhesion between the Ag@cicada wing arrays and Ag nanoparticles. The outstanding optical properties indicate that it has enormous potential in the label-free detection and biological analytes determination.     

Surface‐enhanced Raman scattering on colloid gels originated from low molecular weight gelator

Surface‐enhanced Raman scattering (SERS) on silver and gold colloid gels formed by a low molecular weight organic gelator, bis‐(S‐phenylalanine) oxalyl amide, was obtained. Strong Raman signals dominate in the SERS spectra of hydrogels containing silver nanoparticles prepared by citrate and borohydride reduction methods, whereas broad bands of low intensity are detected in the spectra of gold colloid gels. Resemblance between Raman spectrum of the crystalline substance and the SERS spectra of the silver nanoparticle–hydrogel composites implies the electromagnetic nature of the signal enhancement. A change in Raman intensity of the benzene and amide II bands caused by an increase in temperature and concentration indicates that the gelling molecules are strongly attached through the benzene moieties to the metal nanoparticles while participating in gel formation by intermolecular hydrogen bonding between the adjacent oxalyl amide groups. Transmission electron microscopy reveals a dense gel structure in the close vicinity of the enhancing metal particles for both silver colloid gels. Copyright © 2008 John Wiley & Sons, Ltd.     

基于等离子体腔表面增强拉曼效应的研究

金属离子水滴与液态聚二甲基硅氧烷（PDMS）自发形成的等离子体腔作为一种新型的表面增强拉曼（SERS）基底将等离子体纳米颗粒整合到光学装置中，提高了SERS检测的实用性与可靠性，然而，与其他基底相比，对其最佳生长条件的研究很少。在此，用禁用兽药孔雀石绿（MG）作为探测分子，检验不同生长条件下等离子体腔的特性，包括生长温度和金属离子浓度，以研究等离子体腔的最佳生长条件。金属离子水溶液滴加到互不相容的液态PDMS上时，在表面张力和重力的共同作用下自发形成带开口的球形腔体。同时金属离子扩散到未固化的PDMS中并与残留的Si-H基团反应，金属离子逐渐还原成金属纳米颗粒，并随着PDMS的固化过程在腔体表面逐渐累积，最终形成等离子体腔。其不但能作为角度反射器将入射光限制在腔体中，而且可作为纳米级光子源将吸收的光散射到腔体中，这两个功能共同作用可在基底原本增强作用的基础上进一步提高对MG的拉曼增强效果。较高的生长温度在加快金属离子生长的同时也会加速PDMS的固化，以至于提前结束金属纳米粒子的生长过程。离子浓度越高，形成的金属离子颗粒越大，然而颗粒直径过大，等离子体腔表面的热点数量反而会减少，MG的拉曼增强减弱，因而，必定存在最优化的等离子体腔制备条件使基底对MG的增强效果达到最佳。设置了15，20，25和30 ℃的生长温度以及0.05，0.5，5和50 μg·mL-1的离子浓度，结果表明，在温度为25 ℃，0.5 μg·mL-1的生长条件下等离子体腔实现了对MG的最佳拉曼增强。对等离子体腔生长条件的优化，可为提高该类型基底的SERS增强效果，及可重复制备奠定基础。     

Optical probing and imaging of live cells using SERS labels

During surface‐enhanced Raman scattering (SERS), molecules exhibit a significant increase in their Raman signals when attached, or in very close vicinity, to gold or silver nanostructures. This effect is exploited as the basis of a new class of optical labels. Here we demonstrate robust and sensitive SERS labels as probes for imaging live cells. These hybrid labels consist of gold nanoparticles with Rose Bengal or Crystal Violet attached as reporter molecules. These new labels are stable and nontoxic, do not suffer from photobleaching, and can be excited at any excitation wavelength, even in the near infrared. SERS labels can be detected and imaged through the specific Raman signatures of the reporters. In addition, surface‐enhanced Raman spectroscopy in the local optical fields of the gold nanoparticles also provides sensitive information on the immediate molecular environment of the label in the cell and allows imaging of the native constituents of the cell. This is demonstrated by images based on a characteristic Raman line of the reporter as well as by displaying lipids based on the SERS signal of the C H deformation/bending modes at ∼1470 cm⁻¹. Copyright © 2008 John Wiley & Sons, Ltd.     

纳米Ag材料表面等离子体激元引起的表面增强拉曼散射光谱研究

用热蒸发的方法制备了纳米Ag材料,并用扫描电子显微镜对纳米粒子进行了形貌的表征,通过紫外—可见分光光度计得到Ag纳米粒子的透过谱,得到了Ag纳米粒子的表面等离子体共振的峰值位置.以罗丹明6G为探针分子测定Ag纳米粒子衬底的表面增强拉曼散射效应,通过拉曼散射光谱与透过谱研究了由表面等离子体激元的强极化场引起的表面增强拉曼散射效应,结合透过谱与拉曼增益因子提出了一种描述表面等离子体光学和电学特性的方法,并结合扫描电镜的结果给出了不同结构的纳米Ag材料对表面等离子体激元强度的影响. 关键词： 热蒸发 纳米Ag材料 表面等离子体 表面增强拉曼散射     

Bacterial mixture identification using Raman and surface‐enhanced Raman chemical imaging

The ability of normal Raman and surface‐enhanced Raman scattering (SERS) to identify and detect bacteria has shown great success in recent studies. The addition of silver nanoparticles to bacterial samples not only results in an enhanced Raman signal, but it also suppresses the native fluorescence associated with biological material. In this report, Raman chemical imaging (RCI) was used to analyze individual bacteria and complex mixtures of spores and vegetative cells. RCI uses every pixel or a binned pixel group (BPG) of the Raman camera as an independent Raman spectrograph, allowing collection of spatially resolved Raman spectra. The advantage of this technique resides primarily in the analysis of samples in complex backgrounds without the need for physically isolating or purifying the sample. Using a chemical imaging Raman microscope, we compare normal RCI to SERS‐assisted chemical imaging of mixtures of bacteria. In both cases, we are able to differentiate single bacterium in the Raman microscope's field of view, with a 60‐fold reduction in image acquisition time and a factor of 10 increase in the signal‐to‐noise ratio for SERS chemical imaging over normal RCI. Copyright © 2010 John Wiley & Sons, Ltd.     

Strategy for improved analysis of peptides by surface‐enhanced Raman spectroscopy (SERS) involving positively charged nanoparticles

A strategy for improved surface‐enhanced Raman spectroscopy (SERS) measurements that extends the variety of analytes accessible to SERS analysis is developed. The strategy involves inducing aggregation by mixing positively charged nanoparticles which form SERS‐active clusters when mixed with negatively charged silver nanoparticles fabricated using the Lee–Meisel process. To make positively charged nanoparticles, silver nanoparticles using the traditional Lee–Meisel process are fabricated and coated with a thin layer of silica and the silica modified with silane chemistry. Analytes with a significant amount of negative charge exhibit strong Raman bands when the strategy using these fabricated, positively charged nanoparticles for inducing cluster formation is used, enabling their detection and analysis. We envision the use of positively charged nanoparticles in cluster formation for expanding the range of analytes that can be detected using SERS and hence the range of applications SERS can play a role in. Copyright © 2008 John Wiley & Sons, Ltd.     

AAO模板法制备CdS纳米微粒的SERS光谱研究

在自制的孔径约15nm多孔氧化铝模板上沉积银纳米粒子,然后用电化学方法在此衬底上沉积CdS纳米微粒。研究了CdS纳米阵列在457.5nm波长激光激发下的表面增强拉曼散射(SERS)性质。实验结果显示CdS的SERS信号有三个振动模式,分别对应1LO、2LO和3LO纵光学声子模,它们的强度随着作为SERS衬底的银纳米粒子高度的增加而增强,当银纳米粒子的长/径比(长度与直径的比值)达到4时,这种增强趋近饱和。最后对CdS纳米微粒光学声子模的增强机理进行了分析和讨论。     

Effects of solid substrate on the SERS‐based immunoassay: a comparative study

A comparative study of the solid substrates used in surface‐enhanced Raman scattering (SERS) based immunoassay is made in this paper. Five different substrates were prepared and divided into two groups with and without SERS activity. They are (1) a poly‐L ‐lysine slide, (2) a glutaraldehyde (GA)‐aminosilane slide, (3) a substrate assembled with silver nanoparticles, (4) a substrate assembled with silver nanoparticles and functionalized with GA–aminosilane and (5) a substrate assembled with gold nanoparticles, of which the first two are substrates are without SERS activity and the latter three are with SERS activity because of the existence of the metallic nanoparticles. The SERS experimental results show that the immunoassay performed on an SERS‐active substrate is more effective than that employing the inactive substrate. Among the inactive substrates, the GA–aminosilane slide with a better ability for antibody immobilization leads to a more sensitive immunoassay than the poly‐L ‐lysine slide. Moreover, for SERS‐based immunoassay, the substrate with assembled silver nanoparticles has an advantage of higher SERS enhancement capacity over the substrate assembled with gold nanoparticles. This work indicates that SERS‐active substrates play important and positive roles in sensitive SERS‐based immunoassay. Copyright © 2010 John Wiley & Sons, Ltd.