Background Scleral cross-linking can enhance the biomechanical strength of the sclera and is expected to be a new operative method for the prevention of myopia. However, studies investigating the changes in intraocular pressure (IOP) and ocular pulse amplitude (OPA) after blue light-riboflavin induced scleral collagen cross-linking (SXL) in rhesus monkeys are limited. This study aimed to investigate the changes in IOP and OPA in three-year-old rhesus macaques 1 week, 1 month, and 3 months after blue light-riboflavin SXL. Methods Seven three-year-old rhesus macaques (14 eyes) were randomly divided into two groups, with 4 monkeys in group A (8 eyes) and 3 monkeys in group B (6 eyes). The right eye of each rhesus macaque was used as the experimental eye, whereas the left eye was used as the control. In group A, one quadrant of each right eye was irradiated. In group B, two quadrants of each right eye and one quadrant of each left eye were irradiated. The IOP and OPA of both eyes were measured in all seven rhesus macaques before SXL and 1 week, 1 month, and 3 months postoperatively, and differences in the IOP and OPA between the experimental and control eyes were evaluated via the paired t test. Results In groups A and B, there were no significant differences between the experimental and control eyes in the IOP or OPA before SXL or 1 week, 1 month, or 3 months postoperatively (P > 0.05). Conclusions The IOP and OPA are not significantly affected in 1 vs 0 or in 1 vs 2 quadrants of blue light SXL.