Objective: Work-related discomfort is a pervasive issue among ultrasonography users. The Sonography Work Systems (SWS) framework was constructed as a means of examining relationships within and across components of the sonography work systems, work processes, and work/worker outcomes. A database of ultrasonography users was established as a foundation for a longitudinal survey study to examine worker health and well-being and explore the most salient work systems and process factors associated with work-related discomfort. Materials and Methods: An estimated 100 000 unique ultrasonography users were invited to complete the online questionnaire through an e-mail campaign. Snowball sampling occurred through social media posts and encouragement for respondents to share the survey link with colleagues. The questionnaire included items that examined participant demographics, selected constructs from the SWS, and the prevalence of work-related musculoskeletal discomfort, visual discomfort, and headaches. Individual and multi-factorial regression models were conducted to examine SWS factors associated with the likelihood of experiencing the three types of work-related discomfort. Results: A total of 3659 valid responses were included in the analysis, with 86% of respondents reporting that they regularly experienced musculoskeletal discomfort that they directly attributed to their work. About half (54.2%) of the respondents have engaged in sonography-related ergonomics training, and respondents indicated using adjustable equipment approximately 74% of the time. Workplace culture was rated as primarily positive, but respondents indicated that employers implement only two of seven commonly recommended ergonomic policies and procedures. Working in an organization with more policies, using adjustable equipment more frequently, taking more work breaks, engaging in a positive work culture, and minimizing interruptions to workflow were key factors associated with reduced likelihood of work-related discomfort. Conclusion: This study provides a new framework for examining and addressing factors that contribute to ultrasonography users’ experience of work-related discomfort. Despite increased participation in ergonomics training and the use of adjustable equipment, the prevalence of work-related discomfort remains high among ultrasonography users. The findings highlight the need for attention to be directed at organizational factors and work processes to identify and implement evidence-based solutions to improve the health and well-being of medical ultrasound users.