Aims The primary aim of this study is to assess the reliability of rehabilitative sonography as a non-invasive imaging technique for measuring the visible cross-sectional area (CSA) of the deep suboccipital muscles. Objectives Objectives involve determining both the intra-rater and inter-rater reliability of sonographic measurements to ensure consistent and reproducible results across different sessions and examiners. The ultimate goal is to validate rehabilitative sonography as a reliable tool for clinical and research applications in the assessment and management of musculoskeletal (MSK) conditions involving the deep suboccipital muscles. Design and setting Seventeen participants, including nine women and eight men without neck or vertebral pain, were evaluated using MSK sonography. The visible CSA of suboccipital muscles was assessed using real-time B-mode sonography in a sitting position with a linear probe (5 cm, 7.5 MHz) aligned perpendicularly to the muscle fibers of the rectus capitis posterior minor (RCPM) in one sonogram and the obliquus capitis inferior (OCI) and rectus capitis posterior major (RCPMJ) in another. Two experienced examiners performed sonography measurements, and the procedure was repeated one hour and again one day later. Results Intra-class correlation coefficient (ICC) values for Examiner 1 were 0.84 and 0.79 for RCPM, 0.84 and 0.71 for RCPMJ, and 0.86 and 0.77 for OCI at the one-hour and one-day repetitions, respectively. ICC values for Examiner 2 were 0.86 and 0.77 for RCPM, 0.77 and 0.86 for RCPMJ, and 0.61 and 0.64 for OCI at the one-hour and one-day repetitions, respectively. Inter-rater ICC values were 0.77 for RCPM, 0.82 for RCPMJ, and 0.68 for OCI. Standard error of measurement (SEM) values for Examiner 1 were 11.63 and 13.92 mm² for RCPM, 10.30 and 7.91 mm² for RCPMJ, and 22.84 and 34.61 mm² for OCI. SEM values for Examiner 2 were 11.82 and 13.40 mm² for RCPM, 11.91 and 7.04 mm² for RCPMJ, and 39.20 and 37.73 mm² for OCI. SEM values between examiners were 13.74 mm² for RCPM, 10.36 mm² for RCPMJ, and 36.03 mm² for OCI. Conclusions These findings suggest that sonography is a reliable method for measuring the CSA of the RCPM, RCPMJ, and OCI muscles, provided that the examiners are well-trained and consistent in their techniques and that the average of three measurements is used., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Ethics Committee of University of Social Welfare and Rehabilitation Sciences issued approval IR.USWR.REC.1401.237. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Masoudi et al.)