Cumulative shear strain of the subsynovial connective tissue (SSCT) surrounding finger flexor tendons plays a significant role in the development and progression of carpal tunnel syndrome. Biomechanical risk factors can alter tendon-SSCT shear strain but the effects of external mechanical compression and localized ischemia have yet to be investigated. In a laboratory study with 19 healthy participants, color Doppler ultrasound imaging was used to quantify relative motion between the flexor digitorum superficialis tendon and SSCT during repetitive finger flexion-extension under various conditions of external mechanical compression (palmar and forearm compression), ischemia and different movement speeds (0.75 and 1.25 Hz). Forearm compression reduced tendon displacement (baseline = 28.5 ± 4.1 mm vs. forearm = 27.0 ± 4.6 mm; p = 0.043) and showed a trend for reduced SSCT displacement, while palmar compression had no significant effects on tendon-SSCT motion. Compared with baseline, partial ischemia decreased SSCT displacement (baseline = 22.9 ± 3.3 mm vs. ischemia = 22.0 ± 3.3 mm; p = 0.015), while tendon displacement remained unchanged. In all experimental conditions, faster movements elicited greater tendon-SSCT relative motion. Our findings suggest that palmar compression may not negatively impact tendon-SSCT relative motion, but forearm compression may require further investigation. Localized ischemia in the forearm may alter the gliding conditions within the carpal tunnel and affect tendon-SSCT relative motion, which bridges an important gap between blood flow in the carpal tunnel and shear injury risk. These findings contribute to the growing body of literature, supporting the role that cumulative tendon-SSCT shear injury may have on the pathomechanics of carpal tunnel syndrome.