Objective: To evaluate the significance of CT three-dimensional reconstruction technique for guiding tracheal intubation with rigid fiber bronchoscope in difficult airway. Methods: In this study, 44 patients undergoing selective operation of ASA (American Society of Anesthesiologists physical status) Ⅰ to Ⅱ, neck stiffness, neck trauma needed braked, or severe cervical spondylosis were selected. The patients were randomly divided into two groups: 24 cases in the experimental group (group E) and 20 cases in the control group (group C). The stylets of the rigid fiber bronchoscopes were shaped according to the CT three-dimensional reconstruction images and parameters obtained before surgery. The rigid fiber bronchoscopes shaped according to the CT three-dimensional reconstruction images were used in group E, while the rigid fiber bronchoscopes with the original angles were used in group C. Tracheal intubation operations were all performed by an anesthesiologist who had more than 10 years’ clinical experience and mastered in rigid endoscopic intubation techniques. The first attempt success rate and the total success rate of tracheal intubation, intubation time, blood pressure, heart rate and pulse oxygen saturation at different time points including pre-induction, immediately after intubation, 1-5 minutes after intubation, and intubation related complications within 24 hours were recorded. Results: The total success rate of intubation in the two groups were both 100%. The first attempt success rate of intubation was 96% in group E, and 70% in group C. The first attempt success rate of group E was higher than that of group C. The intubation time of group E was (20.7± 10.6) s, and (21.5 ± 17.6) s of group C. Group E was shorter than that of group C, but there was no significant difference (P > 0.05). RPP equaled the product of heart rate and systolic pressure, which represented the stress reaction of intubation on hemodynamics, was lower in group C at T0, T1, T2, T4 and T5 separately than that in group E, but there was no significant difference (P>0.05). There was no significant difference in tracheal intubation related complications between the two groups (P>0.05). Conclusion: CT three-dimensional reconstruction technique has certain guiding significance in difficult tracheal intubation with rigid fiber bronchoscope in patients with fixed cervical spine.