Abstract: In robot-assisted eye surgery, such as retinal vascular bypass surgery, precise positioning of operating points is required. In this study, a binocular vision-based 3D reconstruction method is proposed to locate the incision points on retinal vessels. Vessels in the image were extracted by CLAHE algorithm to remove the influence of background, then stereo matching was performed. Finally, the retinal vessel image was reconstructed by using the principle of parallax in binocular vision. Experimental results show that this method can accurately locate the incision points on retinal vessels and meet the requirements of ophthalmic surgery.