Respiratory motion during PET has a significant effect on the quantification of radiotracer uptake in PET images. Even when respiratory motion is considered using PET gating techniques, inaccuracies in standardized uptake values can be caused by inappropriate attenuation correction due to a spatial mismatch between PET and CT. In this study, the effect of breath-hold CT imaging on the spatial match between CT and amplitude-based respiratory-gated PET images is investigated. Methods: Wholebody 18 F-FDG PET/CT imaging was performed in 52 patients with 125 lung lesions. 18 F-FDG PET was performed using optimized, amplitude-based respiratory gating. For CT, 36 patients were randomly assigned to the free-breathing (FB) group and 16 to the restexpiratory breath-hold (BH) group. Spatial mismatch between the PET and CT images was quantified by measuring the distance between the centroids of PET and CT lesions and calculating the Jaccard similarity coefficient (JSC). Results: In the upper lobes, the average distance between the centroids of the PET and CT lesions was 4.7 ± 3.1 and 6.0 ± 3.0 mm for the FB and BH groups, respectively (P 5 0.11). For the middle and lower lobes, the distances were 5.8 ± 4.3 and 5.1 ± 2.9 mm (P 5 0.70), respectively, and for the central region 4.8 ± 4.6 and 5.6 ± 2.0 mm (P 5 0.24), respectively. The JSC for the upper lobes was 0.28 ± 0.17 and 0.28 ± 0.19, for the FB and the BH group, respectively (P 5 0.83). For the middle and lower lobes, the JSC was 0.22 ± 0.16 and 0.28 ± 0.18 (P 5 0.20), respectively, and for the central region 0.39 ± 0.17 and 0.13 ± 0.04 (P 5 0.04), respectively. Conclusion: Providing breathing instructions to the patients during the CT acquisition did not improve the spatial alignment between the respiratory-gated PET images and the CT images. The difficulty experienced in using this clinical protocol, such as patient compliance and operator dependence, emphasizes the need for other strategies.Key Words: amplitude-based optimal respiratory gating; lung tumors; standardized uptake value; breath hold CT; spatial alignment PET combined with CT is an essential multimodality molecular imaging method for accurate staging and diagnosis of a variety of diseases, particularly in oncology (1,2). The advantage of combined PET/CT imaging is that it provides both anatomic and molecular information on the patient, improving detection, localization, and characterization of disease (2). Furthermore, the CT scan can be used for PET attenuation correction. Quantitative indices in PET, such as the standardized uptake value (SUV) (3), metabolic volume, and total lesion glycolysis (4,5), can be used to diagnose the disease, to provide prognostic and predictive information, and to optimize radiotherapy planning. Furthermore, it has been established that molecular imaging with PET is valuable in the early assessment of therapy response of several tumor types (6-9).During PET acquisition, patients are instructed to breathe freely because of the relatively long image acquisition times. As a cons...