Instantaneous shifted‐excitation Raman difference spectroscopy (iSERDS)

Shifted‐excitation Raman difference spectroscopy (SERDS) is an experimental method to recover spontaneous Raman spectra despite the presence of strong fluorescence interference. The common scheme requires a tunable laser source and recording two spectra after each other. In this paper, an approach for instantaneous SERDS (iSERDS) is presented utilizing a broadband light source. The broadband radiation is spatially dispersed in the focal plane inside the object of investigation. The generated scattering signal is imaged onto the slit of an imaging spectrograph. The individual pixel lines on the detector represent Raman spectra with slightly shifted excitation wavelength and hence allow SERDS spectra to be derived. The proposed iSERDS technique is a suitable approach for obtaining Raman spectra from fluorescing samples provided they are homogenous on the length scale of the measurement volume. Copyright © 2014 John Wiley & Sons, Ltd.     

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.     

Resonance Raman detection of carotenoid antioxidants in living human tissues

We have used resonance Raman scattering as a novel noninvasive optical technology to measure carotenoid antioxidants in living human tissues of healthy volunteers. By use of blue-green laser excitation, clearly distinguishable carotenoid Raman spectra superimposed on a fluorescence background are obtained. The Raman spectra are obtained within less than a minute, and the required laser light exposure levels are well within safety standards. Our technique can be used for rapid screening of carotenoid levels in large populations and may have applications for assessing antioxidant status and the risk for diseases related to oxidative stress.     

Quantitative analysis of sugar composition in honey using 532‐nm excitation Raman and Raman optical activity spectra

Raman spectroscopy based on the 1064‐nm laser excitation was suggested as a handy non‐invasive technique allowing to quickly determine sugar content in honey and similar food products. In the present study, the green 532‐nm laser radiation is explored instead as it provides higher‐quality spectra in a shorter time. The sample fluorescence was quenched by purification with activated carbon. For control mixture decomposition of Raman spectra to standard subspectra led to a typical error of the sugar content of 3%. Raman optical activity (ROA) spectra that could be measured at the shorter excitation wavelength as well provided a lower accuracy (~8%) than the Raman spectra because of instrumental sensitivity and noise limitations. The results show that Raman spectroscopy provides elegant and reliable means for fast analyses of sugar‐based food products. Copyright © 2016 John Wiley & Sons, Ltd.     

Raman spectroscopy and microscopy of individual cells and cellular components

Raman spectroscopy provides the unique opportunity to nondestructively analyze chemical concentrations in individual cells on the submicrometer length scale without the need for optical labels. This enables the rapid assessment of cellular biochemistry inside living cells, and it allows for their continued analysis. Here, we review recent developments in the analysis of single cells, subcellular compartments, and chemical imaging based on Raman spectroscopy. Spontaneous Raman spectroscopy provides for the full spectral assessment of cellular biochemistry, while coherent Raman techniques, such as coherent anti‐Stokes Raman scattering is primarily used as an imaging tool comparable to confocal fluorescence microscopy. These techniques are complemented by surface‐enhanced Raman spectroscopy, which provides higher sensitivity and local specificity, and also extends the techniques to chemical indicators, i.e. pH sensing. We review the strengths and weaknesses of each technique, demonstrate some of their applications and discuss their potential for future research in cell biology and biomedicine.     

FT-Raman光谱在高分子结构研究中的应用

近年来，继在红外、核磁共振技术领域成功地采用傅立叶变换及计算机技术之后，八十年代后期，傅立叶变换拉曼（ＦＴ－Ｒａｍａｎ）技术突破了原有实验条件的限制，得到了迅速发展［１－５］。九十年代初已进入商品市场。ＦＴ－Ｒａｍａｎ光谱仪的基本结构与普通可见光激光...     

稀土化合物Raman谱中荧光带的辨认

研究了三价稀土氧化物及退火结晶褐钇铌矿和褐铈铌矿在４８８０和５１４５ｎｍ激光激发下所得Ｒａｍａｎ光谱中的荧光带。结果表明,在这两种激光线激发下,Ｓｍ３＋、Ｅｕ３＋和Ｅｒ３＋的荧光对稀土化合物的Ｒａｍａｎ光谱有很大干扰。提出几种在稀土化合物Ｒａｍａｎ光谱中辨认荧光带的方法。     

Background removal from polarized Raman spectra of tooth enamel using the wavelet transform

A wavelet transformation method is introduced to remove the large fluorescence background from polarized Raman spectra of stained tooth enamel. This method exploits the wavelet multiresolution decomposition where the experimental Raman spectrum is decomposed into signals with different frequency components, and where the lowest frequency background and highest frequency noise are removed. This method is optimized using a simulated collection of parallel‐polarized and cross‐polarized Raman spectra of the enamel and then applied to a set of experimental data. The results show that the wavelet transform technique can extract the pure spectra from background and noise, with the depolarization ratio used to discriminate between early dental caries and sound enamel preserved. Copyright © 2010 Crown in the right of Canada. Published by John Wiley & Sons, Ltd.     

Rapid screening of sildenafil and tadalafil adulterated in healthcare products by Micro‐Raman spectroscopy

Sildenafil and tadalafil are inhibitors of phosphodiesterase type 5, which are frequently added into healthcare products. The objective of this study was to evaluate the possibility of using micro‐Raman spectroscopy as a non‐destructive technique to screen for sildenafil and tadalafil in adulterated healthcare products. Using a viewing microscope, the suspect area of healthcare products was selected, which had a discernable crystal form or shape from the surrounding zone. Optimization of instrumental parameters of the Raman spectrometer was chosen to reduce the background fluorescence, and the Raman spectra were collected. The spectra collected were compared with the standard Raman spectra of pure sildenafil and tadalafil. Samples with an identifiable Raman signature to that of sildenafil or tadalafil could be confirmed using liquid chromatography–mass spectrometry (LC/MS). Additionally, wavelet denoising combined with similarity calculation was used to establish an automated approach for discrimination of adulterated healthcare products. Correlation coefficient was chosen for similarity calculation based on the spectra collected and the standard Raman spectra of pure sildenafil and tadalafil. We compared ten samples, secured by administrative authorities in Shanghai, to analyse and demonstrate the capabilities of our proposed method. We established six samples containing sildenafil or tadalafil warranting analysis using LC/MS. Thus, the use of micro Raman spectroscopy provides a quick, convenient and non‐destructive method for screening adulterated chemicals in healthcare products. Raman spectroscopy combined with similarity calculation requires little training after spectra library is developed, thus showing great promise to identify the adulterated healthcare products in the future. Copyright © 2012 John Wiley & Sons, Ltd.     

Approximate chemical analysis of volcanic glasses using Raman spectroscopy

The effect of chemical composition on the Raman spectra of a series of natural calcalkaline silicate glasses has been quantified by performing electron microprobe analyses and obtaining Raman spectra on glassy filaments (~450 µm) derived from a magma mingling experiment. The results provide a robust compositionally‐dependent database for the Raman spectra of natural silicate glasses along the calcalkaline series. An empirical model based on both the acquired Raman spectra and an ideal mixing equation between calcalkaline basaltic and rhyolitic end‐members is constructed enabling the estimation of the chemical composition and degree of polymerization of silicate glasses using Raman spectra. The model is relatively insensitive to acquisition conditions and has been validated using the MPI‐DING geochemical standard glasses 1 as well as further samples. The methods and model developed here offer several advantages compared with other analytical and spectroscopic methods such as infrared spectroscopy, X‐ray fluorescence spectroscopy, electron and ion microprobe analyses, inasmuch as Raman spectroscopy can be performed with a high spatial resolution (1 µm²) without the need for any sample preparation as a nondestructive technique. This study represents an advance in efforts to provide the first database of Raman spectra for natural silicate glasses and yields a new approach for the treatment of Raman spectra, which allows us to extract approximate information about the chemical composition of natural silicate glasses using Raman spectroscopy. We anticipate its application in handheld in situ terrestrial field studies of silicate glasses under extreme conditions (e.g. extraterrestrial and submarine environments). © 2015 The Authors Journal of Raman Spectroscopy Published by John Wiley & Sons Ltd     

Laser Excitation Matrix-Isolation Spectroscopy

Perhaps the most significant recent development in the field of matrix isolation spectroscopy has proven to be the laser-Raman matrix technique. Infrared absorption and Raman scattering spectroscopic methods provide complementary information on vibrational data for the large body of interesting molecular species which require matrix synthesis and stabilization. The laser-excitation technique offers several advantages over the IR experiment: (1) the possibility of obtaining new chemical species by laser photolysis, (2) the observation of resonance Raman spectra which provide harmonic and anharmonic vibrational constants from overtone bands, and (3) the possible observation of fluorescence from new chemical species. Specific experimental examples of complementary IR and Raman studies, laser photolysis, resonance-Raman, and flouorescence spectra are discussed.     

稀土离子光谱探针技术研究——稀土离子与环境配体耦合边带的观测

用纳秒脉冲激光作为激发光源，在激发谱中观察到了Ｔｂ＾３＋的４ｆ－４ｆ电子跃迁与ＯＨ，ＯＤ键伸缩振动的耦合边带。实验上采用了时间分辨技术及扣除背影的计算机数据处理方法，有效地克服 杂质荧光和拉曼散射的背景干扰，获得了清晰的Ｔｂ＾３＋振动谱带。     

紫膜菌紫质的拉曼光谱测试

菌紫质是一种可进行光信息存储与能量转换的生物膜蛋白分子。在光化学循环中 ,它包含的视黄醛产生快速的全反——— 1 3顺式异构化并去质子 ,然后又重新质子化并在反应循环终了转为全反式构型。了解菌紫质的分子构造及光化学反应机制是很有意义的事 ,但由于菌紫质是紫色膜蛋白 ,加之光反应时的荧光问题 ,用普通拉曼技术获得菌紫质的特征峰有一定困难。一般都用时间分辨付里叶变换红外光谱来进行它的测试工作。文本采用传统拉曼技术 ,分别用 5 3 2、785和 1 0 64nm波长激光作激发光源 ,测试了菌紫质的拉曼光谱 ,并得到了菌紫质的大部分特征拉曼谱带。     

Simultaneous Raman micro–spectroscopy of optically trapped and stacked cells

The combination of Raman spectroscopy and optical trapping holds great promise for single‐cell studies and is an emergent theme in microfluidic environments. Here, the evolution of the Raman signal intensity with an axial increment of the mass of the substance of interest inside a specific Raman excitation volume is investigated. Whilst Raman spectroscopy may be applied to tissue samples, solutions and single cells, there are no easily available methods to rapidly acquire signals from small cell populations. We show a simple but powerful method to record the Raman intensity signal simultaneously from a small number of trapped cells or colloidal particles using the technique of optical stacking. The Raman spectra of stacks of red blood cells and yeast cells show that this method can be applied to biological systems. We demonstrate how we may reveal biochemical fingerprints that would otherwise require long integration times for each single cell or averaging over many sequentially acquired cell spectra. There is potential to apply this method to directly attain Raman spectra from sorted sub‐populations of normal, abnormal and tumour cell lines. Copyright © 2007 John Wiley & Sons, Ltd.     

Application of surface‐enhanced Raman spectroscopy (SERS) to the analysis of natural resins in artworks

Surface‐enhanced Raman spectroscopy (SERS) is rapidly growing as an analytical technique for the detection of extremely low concentrations of analytes. The analysis of natural resins from artworks is often restricted by sample size constraints in general, and Raman spectroscopy in particular is hampered by fluorescence when using visible irradiation wavelengths. This work demonstrates that SERS is able to overcome interference from fluorescence in such samples using the incident wavelength 514.5 nm, to allow collection of SERS spectra from extremely small samples. Characterisation of the natural resin surface coating from a painting by Tiepolo is discussed. Copyright © 2008 John Wiley & Sons, Ltd.     

利用特征光谱荧光标记与高斯拟合分析水体中有机物的特性

讨论了一种自然水体中有机物的快速分析方法,以激光诱导荧光(LIF)方法测量了水体的总荧光光谱(TLS);利用特征光谱荧光标记(SFS)技术对水体中溶解有机物(DOM)及叶绿素a(Chl a)的荧光光谱进行了正确的指认和提取。以最小二乘法-Gaussian拟合对水体的TLS进行了拟合,可有效地分离出拉曼散射及荧光光谱,拟合的相关系数优于0.996 4。结果表明,通过在TLS中利用SFS技术正确的指认特定污染物,并以Gaussian拟合进行有效地提取,可以进行水体污染物的快速、实时和在线监测。     

Improved methods for fluorescence background subtraction from Raman spectra

Raman spectroscopy has attracted interest as a non‐invasive optical technique to study the composition and structure of a wide range of materials at the microscopic level. The intrinsic fluorescence background can be orders of magnitude stronger than the Raman scattering, and so, background removal is one of the foremost challenges for quantitative analysis of Raman spectra in many samples. A range of methods anchored in instrumental and computational programming approaches have been proposed for removing fluorescence background signals. An enhanced adaptive weighting scheme for automated fluorescence removal is reported, applicable to both polynomial fitting and penalized least squares approaches. Analysis of the background fitting results for ensembles of simulated spectra suggests that the method is robust and reliable and can significantly improve the background fit over the range of signal, shot noise and background parameters tested, while reducing the subjective nature of the process. The method was also illustrated by application to experimental data generated from aqueous solutions of bulk protein fibrinogen mixed with dextran. Copyright © 2013 John Wiley & Sons, Ltd.     

Raman spectroscopy of proteins: a review

In this work, 26 proteins of different structure, function and properties are investigated by Raman spectroscopy with 488, 532 and 1064 nm laser lines. The excitation lines were chosen in NIR and Vis range as the most common and to show the difference due to normal and resonance effect, sometimes accompanied by the fluorescence. The selected proteins were divided, according to the Structural Classification of Proteins, into four classes according to their secondary structure, i.e. α‐helical (α), β‐sheet (β), mixed structures (α/β, α + β, s) and others. For all compounds, FT‐Raman and two Vis spectra are presented along with the detailed band assignment. To the best of our knowledge, this is the first review showing the potential of Raman spectroscopy for the measurement and analysis of such a large collection of individual proteins. This work can serve as a comprehensive vibrational spectra library, based on our and previous Raman measurements. Copyright © 2013 John Wiley & Sons, Ltd.     

Raman sorting and identification of single living micro-organisms with optical tweezers

We report on a novel technique for sorting and identification of single biological cells and food-borne bacteria based on laser tweezers and Raman spectroscopy (LTRS). With this technique, biological cells of different physiological states in a sample chamber were identified by their Raman spectral signatures and then they were selectively manipulated into a clean collection chamber with optical tweezers through a microchannel. As an example, we sorted the live and dead yeast cells into the collection chamber and validated this with a standard staining technique. We also demonstrated that bacteria existing in spoiled foods could be discriminated from a variety of food particles based on their characteristic Raman spectra and then isolated with laser manipulation. This label-free LTRS sorting technique may find broad applications in microbiology and rapid examination of food-borne diseases.     

菠菜和苏州青类囊体溶液的光谱特性

通过分析不同波长光激发下菠菜(Spinach)和苏州青(Suzhou Green)类囊体溶液的发射光谱和荧光光谱,以及同一波长下磁场处理前后类囊体溶液的光谱变化,探讨了低强度稳恒磁场对类囊体溶液的作用机理.结果表明,类囊体溶液中形成了大量的氢键,磁场处理前菠菜和苏州青的氢键喇曼散射峰为531nm和533.2nm,两者光系统II(PSII)荧光分别在689.8nm和686nm;磁场处理后两者的喇曼散射峰变为532.8nm和532.4nm,PSII荧光变为686.8nm和685.4nm.磁场改变了两样品中氢键键能,使其趋于相等.