Carlo Camerlingo

An Investigation on Micro-Raman Spectra and Wavelet Data Analysis for Pemphigus Vulgaris Follow-up Monitoring.

A wavelet multi-component decomposition algorithm has been used for data analysis of micro-Raman spectra of blood serum samples from patients affected by pemphigus vulgaris at different stages. Pemphigus is a chronic, autoimmune, blistering disease of the skin and mucous membranes with a potentially fatal outcome. Spectra were measured by means of a Raman confocal microspectrometer apparatus using the 632.8 nm line of a He-Ne laser source. A discrete wavelet transform decomposition method has been applied to the recorded Raman spectra in order to overcome problems related to low-level signals and the presence of noise and background components due to light scattering and fluorescence. This numerical data treatment can automatically extract quantitative information from the Raman spectra and makes more reliable the data comparison. Even if an exhaustive investigation has not been done in this work, the feasibility of the follow-up monitoring of pemphigus vulgaris pathology has been clearly proved with useful implications for the clinical applications.

Visible micro-Raman spectroscopy of single human mammary epithelial cells exposed to x-ray radiation.

Abstract. A micro-Raman spectroscopy investigation has been performed in vitro on single human mammary epithelial cells after irradiation by graded x-ray doses. The analysis by principal component analysis (PCA) and interval-PCA (i-PCA) methods has allowed us to point out the small differences in the Raman spectra induced by irradiation. This experimental approach has enabled us to delineate radiation-induced changes in protein, nucleic acid, lipid, and carbohydrate content. In particular, the dose dependence of PCA and i-PCA components has been analyzed. Our results have confirmed that micro-Raman spectroscopy coupled to properly chosen data analysis methods is a very sensitive technique to detect early molecular changes at the single-cell level following exposure to ionizing radiation. This would help in developing innovative approaches to monitor radiation cancer radiotherapy outcome so as to reduce the overall radiation dose and minimize damage to the surrounding healthy cells, both aspects being of great importance in the field of radiation therapy.

Micro-Raman Spectroscopy for Monitoring Changes in Periodontal Ligaments and Gingival Crevicular Fluid

Micro-Raman Spectroscopy is an efficient method for analyzing biological specimens due to its sensitivity to subtle chemical and structural changes. The aim of this study was to use micro-Raman spectroscopy to analyze chemical and structural changes in periodontal ligament after orthodontic force application and in gingival crevicular fluid in presence of periodontal disease. The biopsy of periodontal ligament samples of premolars extracted for orthodontic reasons and the gingival crevicular fluid samples collected by using absorbent paper cones; were analyzed by micro-Raman spectroscopy. Changes of the secondary protein structure related to different times of orthodontic force application were reported; whereas an increase of carotene was revealed in patients affected by periodontal inflammation.

Fructose and Pectin Detection in Fruit-Based Food Products by Surface-Enhanced Raman Spectroscopy

Surface-Enhanced Raman Spectroscopy (SERS) enables the investigation of samples with weak specific Raman signals, such as opaque samples, including fruit juices and pulp. In this paper, biological apple juices and apple/pear pulp have been studied in order to evidence the presence of fructose and pectin, which are components of great relevance for quality assessment of these kinds of products. In order to perform SERS measurements a low-cost home-made substrate consisting of a glass slide decorated with 30-nm-sized gold nanoparticles has been designed and used. By employing a conventional micro-Raman spectroscopy set-up and a suitable data treatment based on “wavelet” denoising algorithms and background subtraction, spectra of pectin and fructose with clear Raman features have been obtained. The results have confirmed the potential of SERS in the food industry for product characterization, also considering the low-cost and the relative ease of the fabrication process of the employed SERS substrate.

Data analysis in Raman measurements of biological tissues using wavelet techniques.

Raman spectroscopy of oral tissues is a promising tool for in vivo diagnosis of oral pathologies, due to the high chemical and structural information content of Raman spectra. However, measurements on biological tissues are usually hindered by low level signals and by the presence of interfering noise and background components due to light diffusion or fluorescence processes. Numerical methods can be used in data analysis, in order to overcome these problems. In this work the wavelet multicomponent decomposition approach has been tested in a series of micro-Raman measurements performed on “in vitro” animal tissue samples. The experimental set-up was mainly composed by a He-Ne laser and a monochromator equipped with a liquid nitrogen cooled CCD equipped with a grating of 1800 grooves/mm. The laser light was focused on the sample surface by means of a 50 X optical objective. The resulting spectra were analysed using a wavelet software package and the contribution of different vibration modes have been singled out. In particular, the C=C stretching mode, and the CH2 bending mode of amide I and amide III and tyrosine contributions were present. The validity of wavelet approach in the data treatment has been also successfully tested on aspirin.

Univariate and Multivariate Analysis of Raman Spectra for Quantitative Determination of Sugars in Beverage Industry

Univariate and multivariate analysis methods have been used for discriminating Raman spectra and determining glucose concentration in glucose aqueous solutions, sweetener mixtures and various commercial sport drinks from Raman spectra in the 600–1,600 cm−1 spectral region. They show well defined and separated vibrational fingerprints of the various contained sugars (glucose, fructose and sucrose). Single component (glucose or sweetener) content was quantified by using an univariate approach and the interval Partial Least Square (iPLS) analysis based on a model built by employing aqueous glucose solutions as reference/calibration samples. The good agreement between these estimations and those from a biochemical assay proves the goodness of the method. These results represent a key step towards the development of a fast, simple, cost-effective Raman-based method for glucose quantification in products of food and beverage industry.

Wavelet data analysis of micro-Raman spectra for follow-up monitoring in oral pathologies

A wavelet multi-component decomposition algorithm has been used for data analysis of micro-Raman spectra from human biological samples. In particular, measurements have been performed on some samples of oral tissue and blood serum from patients affected by pemphigus vulgaris at different stages. Pemphigus is a chronic, autoimmune, blistering disease of the skin and mucous membranes with a potentially fatal outcome. The disease is characterized histologically by intradermal blisters and immunopathologically by the finding of tissue bound and circulating immunoglobulin G (IgG) antibody directed against the cell surface of keratinocytes. More than 150 spectra were measured by means of a Raman confocal microspectrometer apparatus using the 632.8 nm line of a He-Ne laser source. A discrete wavelet transform decomposition method has been applied to the recorded Raman spectra in order to overcome related to low-level signals and the presence of noise and background components due to light scattering and fluorescence. The results indicate that appropriate data processing can contribute to enlarge the medical applications of micro-Raman spectroscopy.

Graphene-Based Raman Spectroscopy for pH Sensing of X-rays Exposed and Unexposed Culture Media and Cells

Graphene provides a unique way of sensing the local pH level of substances on the micrometric scale, with important implications for the monitoring of cellular metabolic activities where proton excretion could occur. Accordingly, an innovative biosensing approach for the quantification of the pH value of biological fluids, to be used also with small amounts of fluids, was realized and tested. It is based on the use of micro-Raman spectroscopy to detect the modifications of the graphene doping level induced by the contact of the graphene with the selected fluids. The approach was preliminarily tested on aqueous solutions of known pH values. It was then used to quantify the pH values of cell culture media directly exposed to different doses of X-ray radiation and to media exposed to X-ray-irradiated cells. The Raman response of cells placed on graphene layers was also examined.

Raman spectroscopy on dentin/resin interface of lased and unlased dental samples

Cavities in human teeth were fabricated by means of Er:YAG laser and compared with samples processed by using conventional drill. The cavities of lased and unlased samples were cured with two different composite resins ( Bis-GMA vs PEX) to investigate differences in their adhesion properties. Morphological characterization of the sample surfaces by SEM analysis was performed. The same samples were then examined by micro-Raman spectroscopy by using He-Ne laser visible light excitation, in order to characterize the dentin/resin interfaces. The Raman spectra exhibited typical peaks of dentine. Resin contributions to the Raman spectra were also detected and have been used to monitor the interface properties of the boundary region. The dependence of the Raman spectrum on spatial position has been monitored. In both lased and unlased samples the PEX diffusion in dentine is limited in a region of order of 10 μm depth. A larger interdiffusion region is observed at the boundary of dentine and Bis-GMA resin.