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<italic>Objective:</italic> This translational study aims to investigate the clinical benefits of indocyanine green (ICG) based near-infrared window II (NIR-II) fluorescence image-guided surgery (FGS) on high-grade glioma (HGG) patients. <italic>Methods:</italic> Patients were randomly assigned to receive FGS or traditional white light image-guided surgery (WLS). The detection rate of NIR-II fluorescence was observed. Complete resection rate, progression-free survival (PFS), overall survival (OS), and neurological status were compared. Tissue samples were obtained from the FGS group, with the diagnosis based on the surgeons and the fluorescence recorded for comparison of diagnostic capability. Patients with WHO grade III gliomas or glioblastomas (GBM) were analyzed separately. <italic>Results:</italic> 15 GBM and 4 WHO grade III glioma patients in the FGS group and 18 GBM and 4 WHO grade III glioma patients in the WLS group were enrolled. The detection rate of NIR-II fluorescence was 100% for GBM. The complete resection rate was significantly increased by the FGS for GBM (FGS, 100% [95% CI 73.41-100] <italic>vs.</italic> WLS, 50% [95% CI 29.03-70.97], <italic>P</italic> = 0.0036). The PFS and OS of the FGS group were also significantly prolonged (Median PFS: FGS, 9.0 months <italic>vs.</italic> WLS, 7.0 months, <italic>P</italic> < 0.0001; Median OS: FGS, 19.0 months <italic>vs.</italic> WLS, 15.5 months, <italic>P</italic> = 0.0002). No recurrence was observed in WHO grade III glioma patients. <italic>Conclusions:</italic> NIR-II FGS achieves a much better complete resection rate of GBM than conventional WLS, leading to greatly improved survival of GBM patients. <italic>Significance:</italic> NIR-II FGS is a highly promising technique worthy of exploring more clinical applications.

Near-Infrared Window II Fluorescence Image-Guided Surgery of High-Grade Gliomas Prolongs the Progression-Free Survival of Patients