Objective To test the feasibility and reproducibility of a handheld ultrasound device (HUD) compared to a standard ultrasound machine for muscle thickness measurements in healthy participants. Methods A prospective cross‐sectional study was designed where two novice operators tested the thickness of the vastus lateralis, rectus femoris, and vastus intermedius muscles on recruited asymptomatic participants with no history of muscle diseases. The anterior–posterior thickness of each muscle was measured three times per operator to evaluate intra‐operator reproducibility and using two machines to evaluate inter‐system reproducibility. Scanning started using the HUD followed by the standard system. Intraclass correlation coefficients (ICC) and simple linear regression were used to test for reproducibility and proportional bias respectively. Results A total of 33 male participants volunteered to take part in this study with a mean age of 22.7 years (6.8). Intra‐operator reproducibility was almost perfect for both operators on both machines (ICC > 0.80). The measurements difference percentage between the machines ranged from 1.8% to 6.6% and inter‐system reproducibility ICC ranged from 0.815 to 0.927 showing excellent reproducibility. Inter‐operator reproducibility was poor to moderate on both machines (ICC: 0.522–0.849). Regression analysis showed no proportional bias in the measurements. All measurements were completed successfully using the HUD. Conclusion The HUD demonstrated excellent accuracy compared to the standard ultrasound machine for measuring thigh muscle thickness.