In the field of wearable robotics, assistance needs to be individualized for the user to maximize benefit. Information from muscle fascicles automatically recorded from brightness mode (B-mode) ultrasound has been used to design assistance profiles that are proportional to the estimated muscle force of young individuals. There is also a desire to develop similar strategies for older adults who may have age-altered physiology. This study introduces and validates a ResNet + 2x-LSTM model for extracting fascicle lengths in young and older adults. The labeling was generated in a semimanual manner for young (40,696 frames) and older adults (34,262 frames) depicting B-mode imaging of the medial gastrocnemius. First, the model was trained on young and tested on both young (R2 = 0.85, RMSE = 2.36 ± 1.51 mm, MAPE = 3.6%, aaDF = 0.48 ± 1.1 mm) and older adults (R2 = 0.53, RMSE = 4.7 ± 2.51 mm, MAPE = 5.19%, aaDF = 1.9 ± 1.39 mm). Then, the performances were trained across all ages (R2 = 0.79, RMSE = 3.95 ± 2.51 mm, MAPE = 4.5%, aaDF = 0.67 ± 1.8 mm). Although age-related muscle loss affects the error of the tracking methodology compared to the young population, the absolute percentage error for individual fascicles leads to a small variation of 3–5%, suggesting that the error may be acceptable in the generation of assistive force profiles.