Exploratory multimodal nonlinear microscopy: Coherent anti-stokes Raman scattering, second harmonic generation, and...

We applied multimodal nonlinear optical microscopy and Raman Spectroscopy to rapidly image Oral Squamous Cell Carcinoma (OSCC), comparing results with Hematoxylin-Eosin (H&E) stained to explore supplementary diagnostic methods.

With ethical approval and patient informed consent, we processed primary tumor resection tissues, preparing frozen sections. Slice 1 was stained with H&E-stained, while Slice 2 was unstained and imaged using Coherent anti-Stokes Raman Scattering (CARS), Second-Harmonic Generation (SHG), Differential Interference Contrast (DIC), and Raman spectroscopy. These were analyzed alongside H&E-stained sections to assess diagnostic capabilities.

Raman spectroscopy revealed distinct differences at 646, 798, 1006, 1091, 1157, 1190, 1450, 1520, and 1660 cm^-1. CARS imaging showed unique cancer nest structures, and SHG highlighted collagen fiber distribution.

Multimodal nonlinear imaging, combining CARS, SHG, and DIC, shows potential as a supplementary diagnostic tool for oral squamous cell carcinoma, aiding in the characterization of pathological features and providing valuable clinical diagnostic information.