Purpose: In dynamic musculoskeletal sonography, probe fixation can contribute to field of view (FOV) consistency, which is necessary for valid analysis of architectural parameters. In this volunteer study, the achieved FOV consistency in fixated ultrasonography was quantified and compared with freehand acquisition. Methods: During five resting periods during cycling exercise, longitudinal B-mode images of the vastus lateralis (VL) muscle were acquired on one thigh with a fixated probe, and by two trained observers on the other thigh. In each acquisition, the structural similarity compared to the first resting period was determined using the complex wavelet structural similarity index (CW-SSIM). Also, the pennation angle of the VL was measured. Both CW-SSIM and pennation angle were compared between fixated and freehand acquisition. Furthermore, the compression of tissue by the probe fixation was measured. Results: In fixated acquisition, a significantly higher structural similarity (p < 0.05) and an improved repeatability of pennation angle measurement were obtained compared to freehand acquisition. Probe fixation compressed muscle tissue by 12% on average. Conclusions: Quantification of the structural similarity showed an increase in FOV consistency with sonography compared to freehand acquisition. The demonstrated feasibility of long-term fixated acquisition might be attractive in many medical fields and sports, and for reduction of work-related ergonomic problems among sonographers.