Oral squamous cell carcinoma is a widespread cancer disease whose survival rate is strongly dependent on early diagnosis and on the degree of the malignancy. The conventional histopathology methods, which are currently the standard ones for diagnosis, are very invasive so that they can be hardly proposed as screening methods for an early and accurate detection of disease. Raman microspectroscopy and Atomic Force Microscopy can be potentially considered useful tools for cancer diagnosis and detection of the malignancy degree because they provide information about biochemical cellular content and nanomechanical properties, respectively, which would be modified by the onset and progression of pathology. The present work shows that both such techniques can successfully discriminate two cellular types of cells characterized by different degree of oral squamous cell carcinoma. The discrimination by Raman microspectroscopy occurs according to a larger content of nucleic acids and minor content of protein component in cells characterized by a larger degree of disease, whereas the discrimination by Atomic Force Microscopy is achieved because of a decrease of stiffness as the degree of disease increase. Overall, both techniques could provide useful diagnostic information related to the degree of malignancy of the oral squamous cell carcinoma disease. Since these techniques need only a few cells from the patients to be investigated, they can be promoted as very interesting tools to be employed in oral cancer screening activities.