A Method for Observation of Benign, Premalignant and Malignant Changes in Clinical Skin Tissue Samples via FT -IR Microspectroscopy

Sunlight causes various types of adverse skin changes on the sun-exposed areas of the skin, in which the most hazardous one is the induction of malignant skin tumours. FT -IR spectra were obtained from specimens excised from normal skin, BCCs, SCCs, MMs, nevi, lesions of solar keratosis and Bowen's disease. Tissue samples from freshly frozen specimens were cut into 2 sections in strictly sequential order to be stained with H & E for histopathological analysis, and then to be air-dried on CaF$_2$ slide glasses for further spectral data acquisition from defined area of interest. Intra- and inter-sample variations were estimated within grouped lesion categories according to each skin component. Mean spectra for each type of tissue pathology in the 800-1800 $cm^{-1}$ / region was interpreted using the classical group frequency approach that showed the most visible differences in spectra of benign, premalignant and malignant changes directly related to protein conformation and nucleic acid bases. The relative intensity of the nucleic acid peak was increased with progression to malignancy. In addition, PCA was able to evaluate and maximise the differences in the spectra by reducing the number of variables characterizing each patient and pathology category. This type of approach to non-destructively estimate the complexity of IR-spectra of inhomogeneous samples such as skin demonstrates the advantage of FT -IR microspectroscopy to be able to observe diseased states (benign, premalignant, malignant) and distinguish them from normal against a huge background of inter- and intra-subject variability.