Purpose Present study compares two routes in nasal passage for transnasal flexible fiberoptic laryngoscopy (TNFFL). Methods Included in the study were 60 patients who were split equally into Group 1 and Group 2 and all underwent TNFFL. The tip of endoscope was introduced between the septum and inferior turbinate along the nasal floor in Group 1 and between the inferior turbinate and middle turbinate in Group 2. O-2 saturation, heart rate, systolic blood pressure, diastolic blood pressure of patients were taken 30 min before and immediately after procedure. Patients were asked to rate the pain-irritation, gag reflex, dyspnea they experienced during procedure using visual analogue scale. Two routes were compared in terms of physiological effects and patient comfort. Results No statistically significant difference was found between two groups in terms of physiological effects or patient comfort. When values recorded immediately after procedure were compared with baseline values, a significant decrease was observed in O-2 saturation and heart rate in Group 1 and Group 2, respectively. No clinically detectable symptoms were observed in the patients. Conclusion Decreases in O-2 saturation and heart rate may be regarded as physiological effects of TNFFL. No significant difference between two routes was noted in the present study in terms of physiological effects and patient comfort during TNFFL. Introducing the endoscope between the inferior and middle turbinates may be a comfortable and safe option, particularly if the endoscope cannot be advanced along the nasal floor due to septum deviation, septal crest, inferior turbinate hypertrophy.