BACKGROUND: Vitreous biopsy is a common technique used to guide management of acute endophthalmitis and help differentiate between infectious and inflammatory conditions. Currently, in-clinic vitreous biopsy is performed with a 25-gauge needle, without the ability to cut vitreous, potentially leading to reduced diagnostic yield. Recent work demonstrated the ability to perform vitreous biopsy with an off-the-shelf vitreous cutter. However, this was limited by complexity of assembly. Here, a technique using a single peel pack vitrectomy cutter is demonstrated for in-clinic vitreous biopsy. METHODS: A 25-gauge vitreous cutter is opened from a peel pack. The drive line is identified, cut to length, and attached to a 10 mL syringe. A 1 mL syringe is attached to the aspiration line. After a trocar is used to place a cannula in the pars plana, the vitreous cutter is introduced into the eye. Cutting is performed by an assistant actuating the 10 mL syringe while the surgeon aspirates from the 1 mL syringe. After sample is collected, antimicrobials are injected if required and the cannula is removed. RESULTS: A peel pack technique simplifies assembly for an in-clinic vitreous biopsy using a manually actuated cutter. CONCLUSION: We present a novel, improved, and simplified technique for vitreous tap using a vitreous cutter provided in a single peel pack, actuated by a single syringe with minimal assembly prior to use. This technique may be more accessible for clinicians than prior techniques and does not require a surgical console.