A retrospective, single-center case-series was initiated to evaluate initial clinical experience with the first commercially available intraoperative optical coherence tomography system (iOCT). The Rescan 700 iOCT (Zeiss, Oberkochen, Germany), a spectral-domain OCT system integrated into a surgical microscope, was used in 40 consecutive cases. A standardized review was used to assess whether iOCT imaging resulted in additional information and/or altered decision-making. The iOCT system was usable in conjunction with common chromovitrectomy dyes and tamponades. The surgeons reported that iOCT imaging provided additional information in 74.1% of the posterior and combined surgical cases, which resulted in altered decision-making in 41.9% of the cases. The iOCT imaging time, on average, amounted to 167 seconds. In anterior procedures, the surgeons reported gaining additional information in 22.2% of all cases, but no cases of altered decision-making were reported. Hereby, the iOCT imaging time amounted to 117 seconds, on average. The demonstrated results suggest iOCT has the potential to improve the quality of posterior and anterior segment surgery.