Multimodal nonlinear microscopy of head and neck carcinoma - toward surgery assisting frozen section analysis

Background Treatment of early cancer stages is deeply connected to a good prognosis, a moderate reduction of the quality of life, and comparably low treatment costs. Methods Head and neck squamous cell carcinomas were investigated using the multimodal combination of coherent anti‐Stokes Raman scattering (CARS), two‐photon excited fluorescence (TPEF), and second‐harmonic generation (SHG) microscopy. Results An increased median TPEF to CARS contrast was found comparing cancerous and healthy squamous epithelium with a p value of 1.8·10−10. A following comprehensive image analysis was able to predict the diagnosis of imaged tissue sections with an overall accuracy of 90% for a 4‐class model. Conclusion Nonlinear multimodal imaging is verified objectively as a valuable diagnostic tool that complements conventional staining protocols and can serve as filter in future clinical routine reducing the pathologist's workload. © 2016 Wiley Periodicals, Inc. Head Neck 38: First–1552, 2016