Using surface‐enhanced Raman scattering (SERS) and fluorescence spectroscopy for screening yeast extracts,a complex component of cell culture media

Yeastolate or yeast extract, which are hydrolysates produced by autolysis of yeast, are often employed as a raw material in the media used for industrial mammalian cell culture. The source and quality of yeastolate can significantly affect cell growth and production; however, analysis of these complex biologically derived materials is not straightforward. The best current method, liquid chromatography–mass spectrometry (LC‐MS), is time‐consuming and requires extensive expertise. This study describes the use of surface‐enhanced Raman scattering (SERS) and fluorescence excitation–emission matrix (EEM) spectroscopy coupled with robust principal component analysis (ROBPCA) for the rapid and facile characterization and discrimination of yeast extracts in aqueous solution. SERS using silver colloids generates time‐dependent signals, where adenine is the strongest contributor, and the spectra are stable and reproducible (< ~3%) at 180 min after mixing. Combining this spectral behavior with chemometric methods enables SERS to be used in discriminating between different yeastolate sources, for assessing lot‐to‐lot variability, and, potentially, to monitor storage‐induced compositional changes. Fluorescence EEM combined with multiway ROBPCA also provides a rapid and inexpensive method for the discrimination of yeastolate, yielding results in terms of sample discrimination very similar to that obtained with SERS. However, the EEM data does not provide the same level of chemical information that is provided by the SERS. Thus, the combination of these two methodologies has the potential to be extremely useful in biopharmaceutical manufacturing, as well as for the rapid characterization and screening of biogenic hydrolysates from animal or plant sources. Copyright © 2011 John Wiley & Sons, Ltd.     

Reorientation dynamics in liquid alcohols from Raman spectroscopy

Polarized Raman spectroscopy has been employed to study the reorientational, or more specifically the translational relaxation dynamics, of alcohol molecules in pure liquids and aqueous solutions. It is found from the spectral width measurements that alcohol molecules in pure liquids have typically translational relaxation times on the order of picoseconds, following the order methanol < ethanol < i‐propanol < n‐propanol. Temperature‐dependent measurements show that hydrogen‐bonding (HB) and hydrophobic interactions control the translational motion. The hydrophobic interaction reduces the relaxation time more apparently in view of the  CH₃ group than the skeleton motion. For alcohol–water mixtures, the increase of water concentration generally slows down the relaxation process in a non‐monotonic behavior. However, the trend stops at a certain point and the motion of alcohol molecules becomes faster when the alcohol concentration further drops. Different mechanisms have been proposed to interpret these observations, which might be helpful to gain deeper insight into the HB networks of alcohols with water. Our study strongly illustrates that Raman spectroscopy can be applied to the study of fast translational motion of molecules in HB systems. Copyright © 2011 John Wiley & Sons, Ltd.     

Raman spectroscopy for study of interplay between phonon confinement and Fano effect in silicon nanowires

A combined effect of doping (type and species) and size on Raman scattering from silicon (Si) nanowires (NWs) has been presented here to study interplay between quantum confinement and Fano effects. The SiNWs prepared from low doping Si wafers show only confinement effect, as evident from the asymmetry in the Raman line‐shape, irrespective of the doping type. On the other hand SiNWs prepared from wafer with high doping shows the presence of electron–phonon interaction in addition to the phonon confinement effect as revealed from the presence of asymmetry and antiresonence in the corresponding Raman spectra. This combined effect induces an extra asymmetry in the lower energy side of Raman peak for n‐type SiNWs whereas the asymmetry flips from lower energy side to the higher energy side of the Raman peak in p‐type SiNWs. Such an interplay can be represented by considering a general Fano‐Raman line‐shape equation to take care of the combined effect in SiNWs. Copyright © 2015 John Wiley & Sons, Ltd.     

Aperture based Raman spectroscopy on SiGe film structures with high spatial resolution

We have investigated silicon–germanium (SiGe) line structures employing metallic apertures in combination with Raman spectroscopy to obtain high‐spatial strain resolution below the diffraction limit. The apertures were cut into specifically shaped electrochemically etched tungsten tips, which were adjusted within the Raman laser beam on the sample surface by a tuning fork atomic force microscope. With this setup, line structures on patterned SiGe films with a center‐to‐center distance down to 200 nm were resolved in the Raman scans, evidently indicating a resolution clearly below the far‐field Raman resolution of about 600 nm for the used instrument. This setup allows improved local strain analysis by Raman spectroscopy and shows potential for further near‐field Raman applications. Copyright © 2008 John Wiley & Sons, Ltd.     

Raman spectroscopy of sol–gel derived titanium oxide thin films

Raman spectra of TiO₂ films prepared via the sol–gel process were studied by UV and visible Raman spectroscopy. The evolution of the phases of TiO₂ films during annealing was investigated, and the relative intensities of the Raman bands excited with 325 nm were found to be distinct from those of the bands excited with 514 nm. The transmittance and FTIR spectra of the films annealed at different temperatures were characterized. The crystallization process of the powders and thin films treated by different annealing methods were also studied with Raman spectroscopy. The results show that the change in the relative intensities is caused by the resonance Raman effect. The anatase to rutile transition of the powder occurs at 700 °C, while that of the thin film occurs at 800 °C. The analysis of Raman band shape (peak position and full width at half‐maximum) after conventional furnace annealing and rapid thermal annealing indicates the influence of the non‐stoichiometry and phonon confinement effect. Copyright © 2011 John Wiley & Sons, Ltd.     

甲苯的激光拉曼光谱研究

采用激光拉曼光谱技术获得了甲苯的振动拉曼光谱,结合群论方法及退偏度测量,指认了其中16条振动谱线,为有机混合物中对甲苯的识别提供了依据．选择甲苯全对称环呼吸振动（1008.8cm－1）为检测谱线,对不同浓度的甲苯／四氯化碳样品进行了分析．研究结果表明：在200．1～2．61g／L浓度范围内,甲苯／四氯化碳振动拉曼光谱强度比与甲苯浓度呈良好的线性关系,线性相关系数为0.997．为甲苯的定量分析提供了一种有效的研究方法．     

单细胞喇曼光谱分析高渗环境对酵母乙醇发酵代谢的影响

应用喇曼光谱和单细胞分析技术监测非发酵底物形成的不同高渗透压对酿酒酵母乙醇发酵的影响,及发酵过程胞内主要生物大分子的变化动态,以期从光谱学角度获知酵母细胞乙醇耐受的分子机制.结果显示,渗透压的升高明显延缓酵母细胞的生长、底物消耗和产物生成,但在2.0mol/L山梨醇下乙醇的最终产量高于对照组.主成分分析显示,不同渗透压主要影响酵母细胞光谱的1 300~1 306和1 443cm-1等源自脂类物质的喇曼峰,说明渗透压影响了胞内脂类物质的合成.主特征峰强度的动态变化显示,高渗透压会显著影响782、1 301、1 602和1 657cm-1峰所表征物质的合成时间和强度,进而影响细胞的代谢方向.结果表明耐高渗菌株能适应高渗环境,调整胞内组分含量,实现高产发酵.     

Quantitative,comparable coherent anti‐Stokes Raman scattering (CARS) spectroscopy: correcting errors in phase retrieval

Coherent anti‐Stokes Raman scattering (CARS) microspectroscopy has demonstrated significant potential for biological and materials imaging. To date, however, the primary mechanism of disseminating CARS spectroscopic information is through pseudocolor imagery, which explicitly neglects a vast majority of the hyperspectral data. Furthermore, current paradigms in CARS spectral processing do not lend themselves to quantitative sample‐to‐sample comparability. The primary limitation stems from the need to accurately measure the so‐called nonresonant background (NRB) that is used to extract the chemically sensitive Raman information from the raw spectra. Measurement of the NRB on a pixel‐by‐pixel basis is a nontrivial task; thus, surrogate NRB from glass or water is typically utilized, resulting in error between the actual and estimated amplitude and phase. In this paper, we present a new methodology for extracting the Raman spectral features that significantly suppresses these errors through phase detrending and scaling. Classic methods of error correction, such as baseline detrending, are demonstrated to be inaccurate and to simply mask the underlying errors. The theoretical justification is presented by re‐developing the theory of phase retrieval via the Kramers–Kronig relation, and we demonstrate that these results are also applicable to maximum entropy method‐based phase retrieval. This new error‐correction approach is experimentally applied to glycerol spectra and tissue images, demonstrating marked consistency between spectra obtained using different NRB estimates and between spectra obtained on different instruments. Additionally, in order to facilitate implementation of these approaches, we have made many of the tools described herein available free for download. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.     

A rapid technique for synthesis of metallic nanoparticles for surface enhanced Raman spectroscopy

Incubator‐shaker method was used as a rapid technique to fabricate an efficient surface enhanced Raman scattering (SERS) substrate by combination of zero valent nanostructures and carbon fiber, which shows dramatic Raman enhancement of nitroaromatic molecule. The fabricated Ag nanoparticle on carbon‐fiber substrate (Ag–C) was used as an efficient SERS substrate to detect the adsorbed 2, 4‐dinitrotoluene molecules with a detection limit of 50 ppm. In advent, our developed SERS substrates could have great potential in detecting other nitro‐aromatic based‐explosive materials, such as 2, 4, 6‐trinitrotoluene molecules. Copyright © 2013 John Wiley & Sons, Ltd.     

VHF-PECVD低温制备微晶硅薄膜的拉曼散射光谱和光发射谱研究

采用拉曼散射光谱和PR650光谱光度计对VHF-PECVD制备的微晶硅薄膜进行了结构表征和在线监测研究.结果表明：功率对材料的晶化率（χc）有一定的调节作用，硅烷浓度大，微调作用更明显；SiH*的强度只能在一定的范围内表征材料的沉积速率，功率大相应的速率反而下降；I［Hα*］/I［SiH*］强度比值反映了材料晶化程度，此结果和拉曼散射光谱测试结果显示出一致性；I［Hβ*］/I［Hα*］的强度比表明氢等离子体中的电子温度随功率的增大而逐渐降低. 关键词： 甚高频等离子体增强化学气相沉积 微晶硅 拉曼散射谱 光发射谱     

A Raman spectroscopic study of teeth affected with molar–incisor hypomineralisation

Raman spectroscopy was used to chemically map lesions associated with molar–incisor hypomineralisation in human teeth. Three teeth with hypomineralised lesions of differing severity, described as white, yellow or brown, were mapped using integral ratios of major component bands (hydroxyapatite, amide I and b‐type carbonate) and principal component analysis scores values. These lesions were found to contain depleted levels of mineral (hydroxyapatite) compared with those of healthy enamel. Principal component analysis also highlighted changes in the phosphate structure and variations in various organic constituents. These variations were consistent with increased disorder in the mineral component of the hypomineralised tooth lesions. Scanning electron microscopy–energy dispersive X‐ray spectroscopy supported the findings based on Raman spectroscopy. Copyright © 2015 John Wiley & Sons, Ltd.     

A method for measuring bending stresses based on electromagnetic actuation and Raman spectroscopy

Bending stress and fracture strength of micromachined structures are the parameters determining the function and life span of microelectromechanical systems (MEMS) and are also indispensable data for MEMS design. This paper reports the results of a study in which a bending stress measurement method based on electromagnetic actuation and Raman spectroscopy has been examined. In this study, an eight‐cantilever mass structure is chosen as the experimental object. The results have indicated that it can accurately measure the bending stresses and fracture strength. Copyright © 2006 John Wiley & Sons, Ltd.     

Structural study of Mg‐bearing sodosilicate glasses by Raman spectroscopy

The presence of magnesium in glasses of geological, medical, and technological interests influences their physicochemical and durability properties. However, the understanding of the role of magnesium is dependent on the combined knowledge of the structural environment of magnesium in the glass or melt and of the silicate network connectivity of the studied systems. In this article, we present a Raman spectroscopic study of the network connectivity of 10 ternary silicate glasses in the system Na₂O MgO SiO₂ and one Mg‐free binary silicate glass Na₂O SiO₂. Results obtained at constant polymerization suggest the existence of various Qⁿ units according to the nature of the modifying cation. As polymerization decreases for Na₂O MgO αSiO₂ glasses (labeled as NMSα with α decreasing from 10 to 2), the band associated with Si O Si bending in fully polymerized region disappears being gradually replaced by a band attributed to Si O Si bending in region containing mainly Q² and Q³ species. For highly polymerized glasses (NMS10‐NMS4), the coexistence of these two bands suggests the presence of two interconnected networks. Concomitantly, the signal associated with Q³ species first increases. For a further decrease of the polymerization, the high wavenumber part of the signal associated with Q³ species decreases, while the intensity of the high wavenumber part of the band related to Q² species increases. This result strongly suggests that magnesium charge‐balances preferentially Q² species rather than Q³ species. Copyright © 2010 John Wiley & Sons, Ltd.     

Visible,near‐infrared,and ultraviolet laser‐excited Raman spectroscopy of the monocytes/macrophages (U937) cells

Raman spectra of the monocytes were recorded with laser excitation at 532, 785, 830, and 244 nm. The measurements of the Raman spectra of monocytes excited with visible, near‐infrared (NIR), and ultraviolet (UV) lasers lad to the following conclusions. (1) The Raman peak pattern of the monocytes can be easily distinguished from those of HeLa and yeast cells; (2) Positions of the Raman peaks of the dried cell are in coincidence with those of the monocytes in a culture cell media. However, the relative intensities of the peaks are changed: the peak centered around 1045 cm⁻¹ is strongly intensified. (3) Raman spectra of the dead monocytes are similar to those of living cells with only one exception: the Raman peak centered around 1004 cm⁻¹ associated with breathing mode of phenylalanine is strongly intensified. The Raman spectra of monocytes excited with 244‐nm UV laser were measured on cells in a cell culture medium. A peak centered at 1485 cm⁻¹ dominates the UV Raman spectra of monocytes. The ratio I₁₅₇₄/I₁₆₁₃ for monocytes is found to be around 0.71. This number reflects the ratio between proteins and DNA content inside a cell and it is found to be twice as high as that of E. coli and 5 times as high as that of gram‐positive bacteria. Copyright © 2009 John Wiley & Sons, Ltd.     

包覆燃料颗粒碳化硅层的Raman光谱研究

本文研究了球床高温气冷堆燃料元件中包覆颗粒碳化硅层的Raman光谱. 通过分析不同制备条件下的碳化硅层断面的Raman光谱的峰位、半高全宽与强度, 明确了包覆颗粒中碳化硅层的晶相特征、密度变化和剩余应力等物性.通过分析不同密度碳化硅层一级Raman峰的同步、异步二维相关谱, 发现其LO模较TO模对于密度变化的响应更敏感. 这些结论为研究球床高温气冷堆燃料元件包覆颗粒中碳化硅层的结构及其物性有指导意义.     

Raman spectroscopy study of damage induced in fluorapatite by swift heavy ion irradiations

Raman spectroscopy was used to study the radiation damage of fluorapatite single crystals and sinters. Krypton and iodine ion irradiations were performed at high energies (∼1 MeV amu⁻¹) for fluences ranging between 1 × 10¹¹ and 5 × 10¹³ cm⁻². Evolution of the symmetric stretching mode of the PO₄³⁻ tetrahedral building blocks (strongest Raman mode observed at 965 cm⁻¹) versus ion fluence was investigated. After irradiation, this peak decreases in intensity and a second broader peak appears at lower wavenumber. The well‐resolved peak has been assigned to the crystalline phase, and the broader one to the amorphous phase. The integrated intensity ratios of these two peaks versus fluence are in good agreement with the damage fractions determined by X‐ray diffraction (XRD). Fits of the amorphous fraction versus fluence show that the amorphization mechanisms is dominated by a single‐impact process for iodine ions and by a double‐impact process for krypton ions in the case of single crystals and sinters. For both irradiations, complete amorphization could not be obtained. The amorphous fraction saturates at a maximum value of 88% for sinters and 72% for single crystals. This is attributed to a recrystallization effect which is more important in single crystals than in sinters. For both types of samples, the crystalline peak shifts slightly to a lower wavenumber with fluence, and then shifts back to its initial value for an amorphous fraction larger than 60%. This feature is attributed to a stress relaxation, as shown in the XRD data, which is accompanied by a decrease of the crystalline peak full‐width at half‐maximum. Copyright © 2011 John Wiley & Sons, Ltd.     

A pigment (CuS) identified by micro‐Raman spectroscopy on a Chinese funerary lacquer ware of West Han Dynasty

A blue pigment was identified by micro‐Raman spectroscopy, X‐ray fluorescence spectroscopy (XRF), scanning electron microscopy (SEM)/energy dispersive X‐ray (EDX) and X‐ray diffraction (XRD). The test sample, a funerary lacquer tray, belongs to West Han Dynasty (206 BC–AD 8) of China and was decorated with faint blue patterns. The result from Raman spectroscopy showed that the faint blue is covellite (CuS) due to the presence of a characteristic peak at 474.5 cm⁻¹, which further was confirmed by XRF, SEM–EDX and XRD. This research indicated that CuS had been used as a blue pigment to decorate lacquer wares from the West Han Dynasty in China. Copyright © 2009 John Wiley & Sons, Ltd.     

Characterization of uranium tetrafluoride (UF4) with Raman spectroscopy

The Raman spectrum of uranium tetrafluoride (UF₄) is unambiguously characterized with multiple Raman excitation laser sources for the first time. Across different laser excitation wavelengths, UF₄ demonstrates 16 distinct Raman bands within the 50–400 cm⁻¹ region. The observed Raman bands are representative of various F–F vibrational modes. UF₄ also shows intense fluorescent bands in the 325–750 nm spectral region. Comparison of the UF₄ spectrum with the ZrF₄ spectrum, its crystalline analog, demonstrates a similar Raman band structure consistent with group theory predictions for expected Raman bands. Additionally, a demonstration of combined scanning electron microscopy and in situ Raman spectroscopy microanalytical measurements of UF₄ particulates shows that despite the inherent weak intensity of Raman bands, identification and characterization are possible for micron‐sized particulates with modern instrumentation. The published well‐characterized UF₄ spectrum is extremely relevant to nuclear materials and nuclear safeguard applications. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Journal of Raman Spectroscopy published by John Wiley & Sons Ltd.     

A shifted‐excitation Raman difference spectroscopy (SERDS) evaluation strategy for the efficient isolation of Raman spectra from extreme fluorescence interference

A biochemical characterization of pathologies in biological tissue can be provided by Raman spectroscopy. Often, the raw spectrum is severely affected by fluorescence interference. We report and compare various spectra‐processing approaches required for the purification of Raman spectra from heavily fluorescence‐interfered raw spectra according to the shifted‐excitation Raman difference spectroscopy method. These approaches cover the entire spectra‐processing chain from the raw spectra to the purified Raman spectra. In detail, we compared (1) area normalization versus z‐score normalization, (2) direct reconstruction of the difference spectra versus reconstruction of zero‐centered difference spectra and (3) collective baseline correction of the reconstructed spectra versus piecewise baseline correction of the reconstructed spectra and, finally, (4) analyzed the influence of the shift of the excitation wavelength on the quality of the reconstructed spectra. Statistical analysis of the spectra showed that – in our experiments – the best results were obtained for the z‐score normalization before subtraction of the normalized spectra, followed by zero‐centering of the difference spectra before reconstruction and a piecewise baseline correction of the pure Raman spectra. With our equipment, a wavelength shift from 784 to 785 nm provided reconstructed spectra of best quality. The analyzed specimens were different tissue types of pigs, tissue from the oral cavity of humans and a model solution of dye dissolved in ethanol. © 2015 The Authors. Journal of Raman Spectroscopy published by John Wiley & Sons Ltd.     

The determination of percentage dissociation of zircon (ZrSiO4) to plasma‐dissociated zircon (ZrO2.SiO2) by Raman spectroscopy

Raman spectroscopy and multivariate calibration techniques are used to determine the percentage conversion of zircon (ZrSiO₄) to plasma‐dissociated zircon, ZrO₂^.SiO₂ (PDZ). The integrated area of a consistent ZrO₂ (monoclinic) Raman band at 477 cm^–1 assigned to the A_g symmetry type from different conversion percentages of the PDZ spectra is used in a multivariate analysis scheme (partial least squares) to develop a predictive model for the subsequent determination of percentage dissociation of zircon to PDZ. In contrast to wet chemical methods, this approach eliminates the use of corrosive acids, e.g. hydrofluoric acid, thus leading to significant reductions in analysis time and material wastage, and it is a quick method that can be used online in a PDZ production facility. These results show best correlation with determination coefficient (R²) values in the range between 99.45 and 99.99% for zircon percentage dissociation in cross‐validation tests. This illustrates not only the versatility of the Raman technique in industrial applications but also the importance of noninvasive testing in the study of zircon and related materials. Copyright © 2011 John Wiley & Sons, Ltd.