To determine the effect of intravitreal injection into different quadrants on pain score and injection comfort. Method: 304 patients were divided into 8 groups that included 38 patients in each. The right and left eyes of the patients were divided into 4 quadrants, superior temporal, inferior temporal, superior nasal, and inferior nasal. Intravitreal injections were performed in the room only used for this purpose and by an experienced ophthalmologist. To eliminate drug-related factors only intravitreal ranibizumab applied patients were included in the study. Injections were performed on one of these quadrants 3.5 mm away from the limbus using 30 gauge needle. All injections were performed under topical proparacaine HCl anesthesia. The severity of the patient's pain after the injection was evaluated by using the visual analog scale. Additionally, the patient's stinging, injection comfort and injection safety were also assessed. Results: 176 (57.9%) of the participants were male and 128 (42.1%) were female. The mean age of all participants was 61,84±12,49 (20-90). Age (p=0,793), gender (p=0,534), pain score (p=0,165), stinging score (p=0,264), patient comfort score (p=0,555), injection safety score (p=0,079) were similar between the groups. Pain (p<0,001) and stinging (p<0,001) scores were higher in females (p<0,001). Patient comfort (p=0,001) and injection safety (p=0,019) scores were lower in females. Conclusions: No difference was found between the quadrants in terms of pain score. Therefore, the physicians may prefer the most comfortable quadrant for themselves and for their patients in intravitreal injection.