Raman spectroscopy has shown great potential in detecting nonmelanoma skin cancer accurately and quickly; however, little direct evidence exists on the sensitivity of measurements to the underlying anatomy. Here, we aimed to correlate Raman measurements directly to the underlying tissue anatomy. We acquired Raman spectra of ex vivo skin tissue from 25 patients undergoing Mohs surgery with a fiber probe. We utilized a previously developed biophysical model to extract key biomarkers in the skin from the Raman spectra. We then examined the correlations between the biomarkers and the major skin structures (including the dermis, sebaceous glands, hair follicles, fat, and two types of nonmelanoma skin cancer—basal cell carcinoma (BCC) and squamous cell carcinoma (SCC)). SCC had a significantly different concentration of keratin, collagen, and nucleic acid than normal structures, while ceramide differentiated BCC from normal structures. Our findings identified the key proteins, lipids, and nucleic acids that discriminate nonmelanoma tumors and healthy skin using Raman spectroscopy. These markers may be promising surgical guidance tools for detecting tumors in resection margins.
