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Optical techniques such as fluorescence and diffuse reflectance spectroscopy are proven to have the potential to provide tissue discrimination during the development of malignancies and hence treated as potential tools for noninvasive optical biopsy in clinical diagnostics. Quantitative optical biopsy is challenging and hence the majority of the existing strategies are based on a qualitative assessment of the concerned tissue. Light–tissue interaction models as well as precise optical phantoms can greatly help in the former and here we present a pilot study to assess the optical properties of a multilayer tissue-specific optical phantom with the help of a database generated using multilayer-Monte Carlo (MCML) models. A set of optical models mimicking the properties of actual and diseased conditions of tissues associated with nonmelanoma skin cancer (NMSC) were devised and MCML simulations of fluorescence and diffuse reflectance were performed on these models to generate the spectral signature of identified biomarkers of NMSC such as hemoglobin, flavin adenine dinucleotide, and collagen. A model library was generated and with the extracted features from modeled spectra, classification of normal and NMSC conditions were tested using the K-nearest neighbor (KNN) classifier. Using an in-house assembled scan-based automated bimodal spectral imaging system with reflectance and fluorescence modalities of operation, a layered, thin, tissue equivalent phantom, fabricated with controlled optical properties mimicking normal and NMSC conditions were tested. The spectral signatures corresponding to the NMSC biomarkers were acquired from this phantom and extracted features from the spectra were tested using the KNN classifier and classification accuracy of 100% was achieved. For further quantitative analysis, the experimental and simulated spectra were compared with respect to the light intensity at the emission peak or absorption dips, spectral line width, and average intensity over a range of wavelength of interest and observed to be analogous within specified and systematic error limits. This methodology is expected to give a better quantitative approach for estimation of tissue properties by correlating the experimental and simulated data.
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