In-office vitreous biopsy is currently performed with a 25-gauge needle or less frequently with a specialized in-office surgical system. This article demonstrates in-office vitreous biopsy with a standard vitreous cutter, using syringes to actuate the cutter. A 79-year-old woman presented six days after intravitreal bevacizumab with endophthalmitis. After subconjunctival anesthesia, a valved 27-gauge trocar was inserted through the pars plana. Two syringes were connected to a pneumatic 27-gauge Alcon vitrectomy handpiece and manually actuated by an assistant while the physician aspirated with a third syringe to obtain the vitreous biopsy. Intravitreal vancomycin and ceftazidime were injected. A total of 0.5 cc of fluid was collected without complications. Manual actuated vitrectomy reliably collects sufficient vitreous samples for diagnostic evaluation and may be safer and more effective than needle biopsy. [Ophthalmic Surg Lasers Imaging Retina 2024;55:116-118.].