Bullous pemphigoid detection by micro-Raman spectroscopy and cluster analysis: Structure alterations of proteins

In this study, micro-Raman spectroscopy was used, for the first time, to detect spectral changes between healthy and diseased skin tissues with bullous pemphigoid (BP). The spectral changes provide information about the biochemical alterations between normal skin and blistered and nonblistered regions in samples diagnosed histopathologically as BP. Raman spectra, characterized by many peaks, revealed the molecular composition of the different skin layers, stratum corneum, epidermis and dermis of normal skin. Comparison of spectra monitored at the dermoepidermal junction (DEJ) of healthy skin with those of blisters caused by BP showed evidence for large variations in the amide I and III regions. The alterations of the protein content, amide I and III, are a result of the appearance of immunoglobulin G (IgG) and fibrin, characteristics of BP. Indication for the disease at early stages was obtained from changes in protein content, evidenced in the measured spectra assisted by cluster analysis. The method employed here can contribute to the nearly real-time diagnosis and to a better understanding of the physical and biomolecular processes effected by BP, and might have implications on other skin diseases.