Research on the Raman spectral character and diagnostic value of squamous cell carcinoma of oral mucosa

Early diagnosis of oral carcinomas is essential for successful treatment. The purpose of the present study is to apply near‐infrared Raman spectroscopy to detect oral squamous cell carcinoma (SCC) and leukoplakia (OLK), in order to establish the diagnostic model of the Raman spectra of oral diseases. We collected Raman spectra of normal, OLK and SCC by near‐infrared Fourier transform Raman spectroscopy. The biochemical variations between different lesions were analyzed by the characteristic bands in the subtracted mean spectra. Gaussian radial basis function support vector machines (SVM) were used to classify spectra and establish the diagnostic models. Major differences were observed in the range between 800 and 1800 cm⁻¹. Compared with normal mucosa, high contents of protein, DNA and lipid in SCC and OLK were observed, but the difference between OLK and normal tissue was not as much as that between normal and SCC. SVM displayed a powerful ability in the classifying of normal and SCC, and the specificity, sensitivity and accuracy were 100, 97.56 and 98.75%, respectively. In discriminating between the OLK and normal groups, the three parameters were 85, 68 and 72.5%. The algorithm showed good ability in grouping and modeling of OLK and SCC, and the three parameters were 95, 97.43 and 96.25%. Combined with SVM, near‐infrared Raman spectroscopy can detect biochemical variations in oral normal mucosa, OLK and SCC, and establish diagnostic models accurately. Copyright © 2009 John Wiley & Sons, Ltd.     

Diagnosis of head and neck squamous cell carcinoma using Raman spectroscopy: tongue tissues

Raman spectroscopy is a powerful optical technique capable of providing the structural information at the molecular level. Thus, the technique can be used to detect biochemical changes associated with carcinogenesis and identify the biomolecules involved in cancer. We studied the Raman spectral characteristics of normal, carcinoma in situ, and invasive squamous cell carcinoma (SCC) tissues of tongue, and identified the spectral features that can discriminate these three tissue types. We found that the intensities of Raman bands assignable to tryptophan increase while those attributable to protein keratin decrease when tissue changes from normal to invasive SCC. The variation observed in the intensity of many discriminating peaks including those of tryptophan and keratin as tissue changes from normal to carcinoma in situ and then to invasive SCC suggests that Raman spectroscopy can be used to monitor progression of the disease. We have also analyzed the data with multivariate statistical methods such as principal component analysis and discriminant function analysis. These chemometric methods clearly separate the whole data into three distinct groups consistent with results of pathology. We were able to detect with 91% success rate the normal and carcinoma in situ tissues and with 89% accuracy the invasive SCC tissues of the tongue. Copyright © 2011 John Wiley & Sons, Ltd.     

Comparison of molecular images as defined by Raman spectra between normal mucosa and squamous cell carcinoma in the oral cavity

Raman spectroscopy has been effectively applied to clinically differentiate normal and cancerous mucosal tissues. Micro‐Raman spectroscopy provides a tool to better understand the molecular basis for the Raman clinical signal. The objective of the current study was to utilize micro‐Raman spectroscopy to define the molecular/spectral differences between normal and abnormal squamous cell carcinoma (SCC) in oral mucosa (in vitro). Understanding this may help in identifying unique spectra or may be useful for in vivo application of this technology. Micro‐Raman (confocal) spectroscopy was used to obtain molecular images of normal and SCC cells of human oral mucosa. Four fresh flashed‐frozen tumor and four matched normal tongue specimens were studied. The spectra covered a wavenumber range from 300 to 4000 cm⁻¹ with a spectral resolution of 8 cm⁻¹ and a spatial resolution of 1.0 µm. The cells were located within thin sections of tongue mucosa biopsies. The excitation wavelength of 515 nm was used. We were able to obtain Raman images with rich information about the spectroscopic and structural features within the cytoplasm, cell membrane, and cell nuclei. Significant spectral differences were observed between the Raman images of normal and malignant squamous cells. The heterogeneity of tumor cells within the abnormal tissue was also demonstrated. Spectral differences demonstrated between both tissue types have provided important information regarding the origins of specific signals within the cells of each tissue type. In our search for specific spectral biomarkers, we believe that a cell surface protein, greatly upregulated in SCC cells, was discovered at 1583 cm⁻¹. Copyright © 2010 John Wiley & Sons, Ltd.     

Acute effects of sono-activated photocatalytic titanium dioxide nanoparticles on oral squamous cell carcinoma

Sonodynamic therapy (SDT) is a new treatment modality using ultrasound to activate certain chemical sensitizers for cancer therapy. In this study, effects of high intensity focused ultrasound (HIFU) combined with photocatalytic titanium dioxide (TiO₂) nanoparticles on human oral squamous cell line HSC-2 were investigated. Viability of HSC-2 cells after 0, 0.1, 1, or 3 s of HIFU irradiation with 20, 32, 55 and 73 W cm⁻² intensities in the presence or absence of TiO₂ was measured immediately after the exposures in vitro. Immediate effects of HIFU (3 s, 73 W cm⁻²) combined with TiO₂ on solid tumors were also examined by histological study. Cytotoxic effect of HIFU + TiO₂ in vitro was significantly higher than that of TiO₂ or HIFU alone with the tendency to increase for higher HIFU intensity, duration, and TiO₂ concentration in the suspension. In vivo results showed significant necrosis and tissue damage in HIFU and HIFU + TiO₂ treated samples. However, penetration of TiO₂ nanoparticles into the cell cytoplasm was only observed in HIFU + TiO₂ treated tissues. In this study, our findings provide a rational basis for the development of an effective HIFU based sonodynamic activation method. This approach offers an attractive non-invasive therapy technique for oral cancer in future.     

Nonlinear spectral imaging of human normal skin,basal cell carcinoma and squamous cell carcinoma based on two-photon excited fluorescence and second-harmonic generation

In this work, we use nonlinear spectral imaging based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) for analyzing the morphology of collagen and elastin and their biochemical variations in basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and normal skin tissue. It was found in this work that there existed apparent differences among BCC, SCC and normal skin in terms of their thickness of the keratin and epithelial layers, their size of elastic fibers, as well as their distribution and spectral characteristics of collagen. These differences can potentially be used to distinguish BCC and SCC from normal skin, and to discriminate between BCC and SCC, as well as to evaluate treatment responses.     

Fluorescence spectral properties of stomach tissues with pathology

Steady-state fluorescence and diffuse reflection spectra are measured for in vivo normal and pathological (chronic atrophic and ulcerating defects, malignant neoplasms) stomach mucous lining tissues. The degree of distortion of the fluorescence spectra is estimated taking light scattering and absorption into account. A combination of Gauss and Lorentz functions is used to decompose the fluorescence spectra. Potential groups of fluorophores are determined and indices are introduced to characterize the dynamics of their contributions to the resultant spectra as pathologies develop. Reabsorption is found to quench the fluorescence of structural proteins by as much as a factor of 3, while scattering of the light can increase the fluorescence intensity of flavin and prophyrin groups by as much as a factor of 2.     

Numerical modelling of spectral,temporal and statistical properties of Raman fiber lasers

Efficient numerical modelling of the power, spectral and statistical properties of partially coherent quasi-CW Raman fiber laser radiation is presented. XPM between pump wave and generated Stokes wave is not important in the generation spectrum broadening and XPM term can be omitted in propagation equation what sufficiently speeds-up simulations. The time dynamics of Raman fiber laser (RFL) is stochastic exhibiting events several times more intense that the mean value on the ps timescale. However, the RFL has different statistical properties on different time scales. The probability density function of spectral power density is exponential for the generation modes located either in the spectrum centre or spectral wings while the phases are distributed uniformly. The pump wave preserves the initial Gaussian statistics during propagation in the laser cavity. Intense pulses in the pump wave are evolved under the SPM influence and are not disturbed by the dispersion. Contrarily, in the generated wave the dispersion plays a significant role that results in stochastic behavior.     

茶氨酸拉曼光谱分析

采用显微共焦拉曼光谱技术测试研究了L-茶氨酸,得到了L-茶氨酸的拉曼谱图,发现在250～1 700和2 800～3 000cm-1能够观测到明显的拉曼信号;通过对谱峰进行归属和分析,得到了不同波数范围内的特征振动模式;其中,在321,900,938,1 153,1 312,1 358,1 454和1 647cm-1找到8个较强的拉曼信号,可作为L-茶氨酸的特征峰。结果表明:拉曼光谱可能成为一种直接、准确和快速的检测茶氨酸的方法。     

Detuning and the Raman spectral gap

Stimulated Raman scattering from a hot inhomogenous plasma is considered with collisional and Landau damping taken into account. For low Z targets absolute Raman scattering is dominant, these resonances being highly sensitive to frequency detuning. This is particularly so at longer wavelengths, resulting in gap in the spectrum. This sensitivity is diminished for high Z targets with the result that significant gain is possible over all wavelengths between the Landau cutoff and twice the wavelength of the laser light.     

Raman spectroscopy in experimental oral carcinogenesis: investigation of abnormal changes in control tissues

Oral cancer is a major cause of mortality in South Asian men owing to rampant tobacco abuse. Cancers are also reported in non‐tobacco habitués, especially women, attributed to chronic irritations from irregular/sharp teeth, improper fillings, and poorly fit dentures. Conventional screening approaches are shown to be effective for high‐risk groups (tobacco/alcohol habitués). Raman spectroscopy (RS) is being extensively explored as an alternate/adjunct tool for diagnosis and management of oral cancers. In a previous Raman study on sequential oral carcinogenesis using hamster buccal pouch model, misclassifications between spectra from control and carcinogen [7,12‐dimethylbenz(a)anthracene (DMBA)]‐treated tissues were observed. Histopathology of some control tissues suggested pathological changes, attributable to repeated forceps‐induced irritations/trauma during animal handling. To explore these changes, in the present study, we recorded spectra from three different types of controls – vehicle control (n = 45), vehicle contralateral (n = 45), and DMBA contralateral (n = 70) – exposed to varying degree of forceps handling, along with DMBA‐treated pouches (n = 70) using a 14‐week carcinogenesis protocol. Spectra certified on the basis of histopathology and abnormal cell proliferation (cyclin D1 expression) were used to build models that were evaluated by independent test spectra from an exclusive set of DMBA‐treated and control animals. Many DMBA‐contralateral, vehicle‐control, and vehicle‐contralateral spectra were identified as higher pathologies, which subsequently corroborated with histopathology/cyclin D1 expression. Repeated forceps‐mediated injuries/irritations, during painting and animal handling, may elicit inflammatory responses, leading to neoplasm. The findings of the study suggest that RS could identify micro‐changes. Further, RS‐based in vivo imaging can serve as a promising label‐free tool for screening even in the non‐habitué population where conventional screening is shown to be not effective. Copyright © 2016 John Wiley & Sons, Ltd.     

Distinguishing between renal oncocytoma and chromophobe renal cell carcinoma using Raman molecular imaging

Raman molecular imaging was evaluated as a tool for differentiating between kidney tumors which are sometimes confused using conventional histopathological methods. Renal oncocytoma and chromophobe renal cell carcinoma are two kidney tumors which occasionally possess overlapping histological features. There is no reliable alternative method to routine histologic examination for differentiating these neoplasms. However, a conclusive diagnosis is important, as renal oncocytoma is benign and chromophobe renal cell carcinoma has malignant potential, requiring different treatment options. In an investigation based on 88 patients, a partial least squares discriminant analysis model generated from Raman molecular images distinguished between chromophobe renal cell carcinoma and renal oncocytoma with a performance of 86% sensitivity and 81% specificity. These initial observations suggest that Raman molecular imaging may be applied not only to the differentiation of these kidney tumors but also to other applications for which conventional histological methods are not sufficient for a conclusive diagnosis. Copyright © 2014 John Wiley & Sons, Ltd.     

Hybrid Rayleigh,Raman and two‐photon excited fluorescence spectral confocal microscopy of living cells

A hybrid fluorescence–Raman confocal microscopy platform is presented, which integrates low‐wavenumber‐resolution Raman imaging, Rayleigh scatter imaging and two‐photon fluorescence (TPE) spectral imaging, fast ‘amplitude‐only’ TPE‐fluorescence imaging and high‐spectral‐resolution Raman imaging. This multi‐dimensional fluorescence–Raman microscopy platform enables rapid imaging along the fluorescence emission and/or Rayleigh scatter dimensions. It is shown that optical contrast in these images can be used to select an area of interest prior to subsequent investigation with high spatially and spectrally resolved Raman imaging. This new microscopy platform combines the strengths of Raman ‘chemical’ imaging with light scattering microscopy and fluorescence microscopy and provides new modes of correlative light microscopy. Simultaneous acquisition of TPE hyperspectral fluorescence imaging and Raman imaging illustrates spatial relationships of fluorophores, water, lipid and protein in cells. The fluorescence–Raman microscope is demonstrated in an application to living human bone marrow stromal stem cells. Copyright © 2009 John Wiley & Sons, Ltd.     

Integrated spectral and spatial information extraction in Raman spectroscopy

Raman spectroscopy has been widely used to analyze various substances quantitatively. Conventional studies are primarily focused on the spectral characteristics of Raman scattering. The spatial distribution is always ignored, which can be used to observe the physical properties, such as the particle size. In this article, the spatial information has been extracted from the Raman spectra of barium nitrate, demonstrating that the evident spatial width broadening varied with the particle size. The numerical result shows that the spatial width has a better linear correlation with the particle size, while the Raman intensity has a poor linear correlation. The integrated spectral and spatial information extracted in Raman spectroscopy has a potential application in the quantitative analysis of physical properties.     

磁几何阻挫材料天然绿盐铜矿的中红外和拉曼光谱分析

在室温情况下,利用FTIR光谱仪测得了天然磁几何阻挫材料绿盐铜矿(Atacamite,β-Cu2 (OH)3Cl)的中红外光谱4 000～400 cm-1,同时还通过激光显微拉曼光谱仪获得了它的拉曼光谱4 000～95 cm-1.结合它的晶体结构参数,确认了官能团区4 000～2 500～1 000 cm-1和指纹区1...     

Origin of spectral interferences in femtosecond stimulated Raman microscopy

In femtosecond stimulated Raman microscopy (FSRM), a spectrally narrow (Raman pump) pulse and a broad (Raman probe) laser pulse are employed to generate the Raman spectra of microscopic objects. The resulting spectra exhibit, in addition to the Raman bands, spectral modulations of comparable amplitude. Here a model is devised that attributes these modulations to a four‐wave mixing (FWM) process. Two light fields of the probe pulse and one field of the pump pulse serve as input fields. The resulting FWM field experiences a heterodyne amplification by the probe field. Simulations based on this model reproduce the appearance of the spectral modulations. Furthermore, the amplitude of the modulations exhibits dependences on the energies of pump and probe pulses as well as on the nonlinear refractive index n₂, which are in line with the model. Copyright © 2011 John Wiley & Sons, Ltd.     

Raman spectral imaging of a carbon nanotube intramolecular junction

Confocal Raman spectral imaging results are presented for a metal-to-semiconductor intramolecular junction (IMJ) on an isolated carbon nanotube. Spectra observed at the junction are consistent with the symmetry lowering expected from the occurrence of pentagon-heptagon defects to produce the chirality shift. The IMJ transition zone is sharp and preserves the nanotube diameter. These results have significant implications for the future use of IMJs as electronic devices, including how prevalent these structures are and how their growth may be rationally targeted. Raman imaging has been demonstrated to be a powerful tool for IMJ studies and provides a more accessible method for further studies of IMJ structure and growth.     

Apoptosis in tongue squamous cell carcinoma and its correlation with clinically occult cervical metastasis

Squamous cell carcinoma of the tongue is one of the most common head and neck cancer. Treatment in the early stages is still controversial. This study aims to correlate the apoptosis rate of primary tongue cancer with cervical node metastasis, found in the histopathological studies of specimens obtained from neck dissections or during clinical follow-up. Twenty patients, 65% males, mean age 64.5 years, without clinical evidence of cervical metastasis was included. Eighty-five percent were smokers and 40% were alcohol abusers. Mann-Whitney test was used for statistical analysis (p相似文献   

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.