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Background: The exoscope has been proposed as a valid tool in 5-aminolevulinic acid–guided resection of high-grade gliomas. However, it is not clear if, beyond ergonomics, the exoscope provides a real benefit over the optical microscope (OM). The aim of this study was to compare the exoscope with the OM in terms of surgical visualization and workflow in 5-aminolevulinic acid–guided brain surgery. Methods: Surgical videos of patients diagnosed with histopathologically confirmed, Shinoda stage I, high-grade gliomas who underwent surgery in from January to April 2022 were studied. Visualization under a 5-aminolevulinic acid blue filter for vessels, parenchyma, surgical instruments, and fluorescence was categorized for both superficial and deep fields. The following data were also recorded: median number of switches between white light and blue filter, average duration per switch, and amount of work under blue filter. Results: There were 5 surgeries performed under OM guidance and 5 performed under exoscope guidance. Under a blue filter, the exoscope was significantly better than the OM in visualizing vessels, parenchyma, and surgical instruments for both superficial and deep surgical fields. The median number of switches between blue and white light was lower compared with the OM. Both median switch duration and percentage of work under the blue filter were superior when using the exoscope. Conclusions: Within the limitations of a preliminary analysis, use of the exoscope in fluorescence-guided surgery for high-grade gliomas provided significant advantages in terms of visualization of the surgical field under a blue filter and linearity of surgical flow.