The Tono-Pen XL (Bio-Rad, Santa Ana, CA) was compared with manometer readings for intraoperative measurement of intraocular pressure (IOP) in 40 eyes of 40 consecutive patients after vitrectomy, lensectomy, and fluid-air exchange. Tono-Pen readings corresponding to manometer readings of 10, 20, 30, 40, and 50 mmHg were obtained in a masked fashion with a randomized sequence of manometer readings. A correlation was obtained between the manometer and Tono-Pen readings (r = 0.96 in emmetropic eyes and r = 0.93 in myopic eyes). The regression curve that represents the calibration curve of Tono-Pen in terms of the manometer readings for air-filled vitrectomized eyes was obtained. Any Tono-Pen reading can be easily translated into the corresponding manometer reading by referring to the curve. The Tono-Pen can therefore be effectively used to accurately determine intraoperative IOP in eyes undergoing vitrectomy, lensectomy, and fluid-air exchange.