Stimulated Raman histology for histological evaluation of oral squamous cell carcinoma

Abstract

Objectives: To investigate whether in patients undergoing surgery for oral squamous cell carcinoma, stimulated Raman histology (SRH), in comparison with H&E-stained frozen sections, can provide accurate diagnoses regarding neoplastic tissue and sub-classification of non-neoplastic tissues.

Materials and methods: SRH, a technology based on Raman scattering, was applied to generate digital histopathologic images of 80 tissue samples obtained from 8 oral squamous cell carcinoma (OSCC) patients. Conventional H&E-stained frozen sections were then obtained from all 80 samples. All images/sections (SRH and H&E) were analyzed for squamous cell carcinoma, normal mucosa, connective tissue, muscle tissue, adipose tissue, salivary gland tissue, lymphatic tissue, and inflammatory cells. Agreement between SRH and H&E was evaluated by calculating Cohen's kappa. Accuracy of SRH compared to H&E was quantified by calculating sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) as well as area under the receiver operating characteristic curve (AUC).

Results: Thirty-six of 80 samples were classified as OSCC by H&E-based diagnosis. Regarding the differentiation between neoplastic and non-neoplastic tissue, high agreement between H&E and SRH (kappa: 0.880) and high accuracy of SRH (sensitivity: 100%; specificity: 90.91%; PPV: 90.00%, NPV: 100%; AUC: 0.954) were demonstrated. For sub-classification of non-neoplastic tissues, SRH performance was dependent on the type of tissue, with high agreement and accuracy for normal mucosa, muscle tissue, and salivary glands.

Conclusion: SRH provides high accuracy in discriminating neoplastic and non-neoplastic tissues. Regarding sub-classification of non-neoplastic tissues in OSCC patients, accuracy varies depending on the type of tissue examined.

Clinical relevance: This study demonstrates the potential of SRH for intraoperative imaging of fresh, unprocessed tissue specimens from OSCC patients without the need for sectioning or staining.

Keywords: Microscopy; Oral squamous cell carcinoma; Pathology; Raman spectroscopy; Surgical margins.