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To evaluate intraocular pressure (IOP) fluctuation during vitrectomy, we directly monitored IOP in vivo using 2 vitrectomy machines with or without constant infusion pressure monitoring and control.Among 61 eyes of 61 consecutive patients, 32 were assigned to the Accurus system (group 1) and 29 were assigned to the Constellation system (group 2) in this prospective case series. The IOP fluctuations were evaluated during routine vitrectomy procedures.The initial IOP before vitrectomy was 20.3 ± 2.4 mm Hg in group 1 using a conventional vented gas forced infusion system and 20.0 ± 0.0 mm Hg in group 2 using active IOP control at 20 mm Hg (p = 0.532). However, the average IOP change during core vitrectomy was -8.6 ± 4.3 mm Hg in group 1 and -0.8 ± 1.1 in group 2 (p<00.001). Maximum IOP was significantly decreased in group 1 (-17.0 ± 2.6 mm Hg) compared with that in group 2 (-4.1 ± 2.2 mm Hg) (p<00.001). Partial ocular collapse was observed during vitrectomy only in group 1 (78.1%). Peak IOP significantly increased during scleral compression and gas and fluid injection but was not significantly different between the groups (all p≥0.147). The IOP fluctuation range was 50-70 mm Hg in both groups.The IOP fluctuated significantly during routine vitrectomy using both systems. Hypotony and partial ocular collapse were more frequently observed during vitrectomy with the Accurus system than with the Constellation system. Both systems were vulnerable to IOP surge during indentation and intravitreal injection.