Exploratory multimodal nonlinear microscopy: Coherent anti-stokes Raman scattering, second harmonic generation, and differential interference contrast for oral squamous cell carcinoma diagnosis

Abstract

Objective

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.

Study Design

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.

Results

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.

Conclusions

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.