Raman spectroscopic analysis of oral cells in the high wavenumber region (original) (raw)
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Biophotonics South America, 2015
Raman spectroscopy can provide a molecular-level signature of the biochemical composition and structure of cells with excellent spatial resolution and could be useful to monitor changes in composition for early stage and non-invasive cancer diagnosis, both ex-vivo and in vivo. In particular, the fingerprint spectral region (400-1,800 cm-1) has been shown to be very promising for optical biopsy purposes. However, limitations to discrimination of dysplastic and inflammatory processes based on the fingerprint region still persist. In addition, the Raman spectral signal of dysplastic cells is one important source of misdiagnosis of normal versus pathological tissues. The high wavenumber region (2,800-3,600 cm-1) provides more specific information based on N-H, O-H and C-H vibrations and can be used to identify the subtle changes which could be important for discrimination of samples. In this study, we demonstrate the potential of the highwavenumber spectral region by collecting Raman spectra of nucleoli, nucleus and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (DOK) cell lines and from normal oral epithelial primary cells, in vitro, which were then analyzed by area under the curve as a method to discriminate the spectra. In this region, we will show the discriminatory potential of the CH vibrational modes of nucleic acids, proteins and lipids. This technique demonstrated more efficient discrimination than the fingerprint region when we compared the cell cultures.
Ex vivo analysis of the oral epithelium by high-wavenumber Raman spectroscopy
International Journal of Biomedical Engineering and Technology, 2017
Raman spectroscopy at wavenumber region of 400-1800 cm-1 is an already proved promising technology for objective clinical evaluation of biological tissues to obtain their molecular fingerprint. The study of the highwavenumber region (2800-3100 cm-1) is also of interest to significantly shorten the diagnosis time due to the intense Raman signal in a narrower range. This work studies parameters dependence, Raman saturation in depth and repeatability of tissue measurements at this region and proposes three indices for quantitative diagnose in oral epithelium. Sixteen oral mucosa biopsies in different thicknesses were evaluated in varying laser powers. The nonsaturation of Raman signal up to 120 µm in depth was demonstrated. Three peaks (2874, 2926, 3056 cm-1) attributed to functional groups of interest in oral mucosa were identified from the Raman band deconvolution. The highwavenumber correspondence with the fingerprint region (400-1800 cm-1) was proven and three diagnose indices proposed to quantitatively evaluate oral epithelium tissue in routinely clinical practice.
Raman Spectroscopy Analysis for Optical Diagnosis of Oral Cancer Detection
Journal of Clinical Medicine
Raman spectroscopy (RS) is widely used as a non-invasive technique in screening for the diagnosis of oral cancer. The potential of this optical technique for several biomedical applications has been proved. This work studies the efficacy of RS in detecting oral cancer using sub-site-wise differentiation. A total of 80 samples (44 tumor and 36 normal) were cryopreserved from three different sub-sites: The tongue, the buccal mucosa, and the gingiva of the oral mucosa during surgery. Linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA) were used with principal component analysis (PCA) to classify the samples and the classifications were validated by leave-one-out-cross-validation (LOOCV) and k-fold cross-validation methods. The normal and tumor tissues were differentiated under the PCA-LDA model with an accuracy of 81.25% (sensitivity: 77.27%, specificity: 86.11%). The PCA-QDA classifier model differentiated these tissues with an accuracy of 87.5% (sensitivity: ...
Near-Infrared Raman Spectroscopy for Oral Carcinoma Diagnosis
Photomed Laser Surg, 2006
Raman spectroscopy has been applied as a diagnostic tool for the detection of cancers, due to its sensitivity to the changes in molecular composition and conformation that occurs in malignant tissues. The detection of weak Raman signals from biotissues becomes easier by FT-Raman due to fluorescence suppression. Methods: A carcinogen (7,12-dimethybenz[a]anthracene [DMBA]) was applied daily in the oral pouch of 21 hamster to induce oral carcinoma. After 14 weeks, the fragments of squamous cell carcinomas and oral normal tissue were collected and analyzed by FT-Raman spectroscopy, using a 1064-nm Nd:YAG laser line as an excitation source. A total of 123 spectra were obtained and divided in normal and malignant tissue groups, and analyzed statistically through principal components analysis (PCA) and classified using Mahalanobis distance. Results: Major differences between normal and malignant spectra seem to arise from the composition, conformational, and structural changes of proteins, and possible increase of its content in malignant epithelia. An algorithm based on PCA was able to separate the samples into two groups-normal and carcinoma. For the algorithm training group, 91% sensitivity and 69% specificity were observed, while the prospective group had 100% sensitivity and 55% specificity. Conclusion: The algorithm based on PCA has the potential for classifying Raman spectra and can be useful for detection of dysplastic and malign oral lesion. 348
Raman spectroscopy of normal oral buccal mucosa tissues: study on intact and incised biopsies
Journal of Biomedical Optics, 2011
Oral squamous cell carcinoma is one of among the top 10 malignancies. Optical spectroscopy, including Raman, is being actively pursued as alternative/adjunct for cancer diagnosis. Earlier studies have demonstrated the feasibility of classifying normal, premalignant, and malignant oral ex vivo tissues. Spectral features showed predominance of lipids and proteins in normal and cancer conditions, respectively, which were attributed to membrane lipids and surface proteins. In view of recent developments in deep tissue Raman spectroscopy, we have recorded Raman spectra from superior and inferior surfaces of 10 normal oral tissues on intact, as well as incised, biopsies after separation of epithelium from connective tissue. Spectral variations and similarities among different groups were explored by unsupervised (principal component analysis) and supervised (linear discriminant analysis, factorial discriminant analysis) methodologies. Clusters of spectra from superior and inferior surfaces of intact tissues show a high overlap; whereas spectra from separated epithelium and connective tissue sections yielded clear clusters, though they also overlap on clusters of intact tissues. Spectra of all four groups of normal tissues gave exclusive clusters when tested against malignant spectra. Thus, this study demonstrates that spectra recorded from the superior surface of an intact tissue may have contributions from deeper layers but has no bearing from the classification of a malignant tissues point of view.
Raman Micro-Spectroscopy for Rapid Screening of Oral Squamous Cell Carcinoma
Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology, 2015
Raman spectroscopy can provide a molecular-level fingerprint of the biochemical composition and structure of cells with excellent spatial resolution and could be useful to monitor changes in composition for dysplasia and early, non-invasive cancer diagnosis (carcinoma in situ), both ex-vivo and in vivo. In this study, we demonstrate this potential by collecting Raman spectra of nucleoli, nuclei and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (pre-cancerous, DOK) cell lines and from normal oral epithelial primary cell cultures, in vitro, which were then analysed by principal component analysis (PCA) as a multivariate statistical method to discriminate the spectra. Results show significant discrimination between cancer and normal cell lines. Furthermore, the dysplastic and cancer cell lines could be discriminated based on the spectral profiles of the cytoplasmic regions. The principal component loading plot, which elucidates the biochemical features responsible for the discrimination, showed significant contributions of nucleic acid and proteins for nucleolar and nuclear sites and variation in features of lipids for the cytoplasmic area. This technique may provide a rapid screening method and have potential use in the diagnosis of dysplasia and early, non-invasive oral cancer, the treatment of which involves much less extensive and complex surgery and a reduction in associated co-morbidity for the patient.
Investigation of the potential of Raman spectroscopy for oral cancer detection in surgical margins
Laboratory Investigation, 2015
The poor prognosis of oral cavity squamous cell carcinoma (OCSCC) patients is associated with residual tumor after surgery. Raman spectroscopy has the potential to provide an objective intra-operative evaluation of the surgical margins. Our aim was to understand the discriminatory basis of Raman spectroscopy at a histological level. In total, 127 pseudocolor Raman images were generated from unstained thin tissue sections of 25 samples (11 OCSCC and 14 healthy) of 10 patients. These images were clearly linked to the histopathological evaluation of the same sections after hematoxylin and eosin-staining. In this way, Raman spectra were annotated as OCSCC or as a surrounding healthy tissue structure (i.e., squamous epithelium, connective tissue (CT), adipose tissue, muscle, gland, or nerve). These annotated spectra were used as input for linear discriminant analysis (LDA) models to discriminate between OCSCC spectra and healthy tissue spectra. A database was acquired with 88 spectra of OCSCC and 632 spectra of healthy tissue. The LDA models could distinguish OCSCC spectra from the spectra of adipose tissue, nerve, muscle, gland, CT, and squamous epithelium in 100%, 100%, 97%, 94%, 93%, and 75% of the cases, respectively. More specifically, the structures that were most often confused with OCSCC were dysplastic epithelium, basal layers of epithelium, inflammation-and capillary-rich CT, and connective and glandular tissue close to OCSCC. Our study shows how well Raman spectroscopy enables discrimination between OCSCC and surrounding healthy tissue structures. This knowledge supports the development of robust and reliable classification algorithms for future implementation of Raman spectroscopy in clinical practice.
The Potential of Raman Spectroscopy in the Diagnosis of Dysplastic and Malignant Oral Lesions
Cancers
Early diagnosis, treatment and/or surveillance of oral premalignant lesions are important in preventing progression to oral squamous cell carcinoma (OSCC). The current gold standard is through histopathological diagnosis, which is limited by inter- and intra-observer errors and sampling errors. The objective of this work was to use Raman spectroscopy to discriminate between benign, mild, moderate and severe dysplasia and OSCC in formalin fixed paraffin preserved (FFPP) tissues. The study included 72 different pathologies from which 17 were benign lesions, 20 mildly dysplastic, 20 moderately dysplastic, 10 severely dysplastic and 5 invasive OSCC. The glass substrate and paraffin wax background were digitally removed and PLSDA with LOPO cross-validation was used to differentiate the pathologies. OSCC could be differentiated from the other pathologies with an accuracy of 70%, while the accuracy of the classifier for benign, moderate and severe dysplasia was ~60%. The accuracy of the cl...
Raman spectroscopic characterization of blood plasma of oral cancer
2013 IEEE 4th International Conference on Photonics (ICP), 2013
Raman Spectroscopy is a versatile technique to probe in to the vibrational or rotational transitions of a molecule and extract complete information about the biochemical composition of the sample under investigation. The metabolic end products of the cell that were released in to the circulating blood would change the biological molecules and thus alter their spectral signatures.