Objective To analysis the correlation among the intraocular pressure readings of Tono-Pen obtained from various corneal and scleral locations. Methods The intraocular pressure of forty New Zerland white rabbits (eighty eyes) were examined with Tono-Pen and schiotz tonometer under general anesthesia. Four different locations of Each eye were taken with Tono-Pen tonometer, that is central cornea, pericentral cornea, limbus and sclera four minimeters behind the limbus. The IOP of central cornea were also taken with schiotz tonometer The data were analyzed with SPSS software. Results The Tono-Pen readings obtained from various locations were (19.44±2.33)mmHg、(19.44±1.95)mmHg、(21.66±4.26)mmHg、(21.83±4.84)mmHg separately. There was correlation between the IOP of pericentral cornea and central cornea (r=0.615, P=0.007); While the correlation rate between the IOP of limbus and central cornea was 0.537 (P=0.02), the correlation rate between the IOP of between sclera four minimeters behind the limbus and central cornea was 0.215 (P=0.391). Conclusion The IOP taken by Tono-Pen tonometer of pericentral cornea or limbus may be the real IOP of rabbit.