Purpose: Endoscopic third ventriculostomy is an established method for treating hydrocephalus. The third ventriculostomy site is considered a safe area that can be disrupted during surgical endoscopic procedures. The question of the clinical consequences of an apparently unavoidable injury to the floor of the third ventricle has been sporadically addressed in the literature. The aim of this study is to describe our anatomical and operative findings during endoscopic procedures performed in fluorescent mode after intravenous fluorescein injection and address the possible role of fluorescein-enhanced visualization of the median eminence as an accessory tool in order to partially spare this functional structure when performing ventriculostomy. Methods: We prospectively administered intravenously 500 mg of fluorescein sodium in 12 consecutive endoscopic surgery cases. A flexible scope equipped with dual observation modes for both white light and fluorescence was used. Taking into account the position of the basilar apex and the need for a conveniently sized stoma, a perforation area was chosen and dilated using a Fogarty balloon, guided by fluorescein-enhanced visualization of the median eminence. Results: After a mean of 20 s in the fluorescent mode, the fluorescein enhanced the visualization of the median eminence-tuber cinereum complex. In our preliminary experience, by opening the stoma in the fluorescence mode, almost half of the visible median eminence surface can be spared from iatrogenic sacrifice. Conclusions: Tailoring fluorescence-guided ventriculostomy is a feasible way of trying to preserve the median eminence and may have implications for the site and safety of this common surgical procedure