Current image-guided neuronavigation techniques, such as intraoperative MRI and ultrasound, are limited by high costs, limited sensitivity and operational complexity. There is a clear need for more efficient systems to enhance intra-operative tumor detection and improve patient outcomes for intrinsic brain tumor resection. Whereas previous intraoperative brain HSI studies have predominantly investigated high-grade gliomas, glioblastomas, or mixed-grade cohorts, this clinical study focuses specifically on low-grade gliomas (LGGs). LGGs strongly resemble normal brain under white light conditions on the operative microscope. This study characterizes spectral differences between healthy and tumor tissue and provides a paired comparison of HSI- and RGB-based classification at matched tissue locations and under identical patient-level data splits. Tumor regions showed consistently lower reflectance, which may be associated with differences in hemoglobin concentration and oxygenation. A range of classification models was evaluated. The 3D CNN, which integrates spectral and spatial features, achieved the highest median ROC-AUC of 74.5%, compared with 67.9% for the spectral-only XGBoost model and 64.0% for the RGB-based model. These results support further investigation of HSI for low-grade glioma classification and quantify its performance relative to conventional RGB imaging in this cohort.

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