Background: Ambulation devices may differ in their utility, muscle activation patterns, and how they affect regional blood flow. This study aimed to evaluate popliteal blood flow and vessel dimensions in response to ambulation with a hands-free crutch (HFC), axillary crutches (AC), a medical kneeling scooter (MKS), and regular walking in healthy adults. Methods: HFC, AC, MKS, and regular walking were completed in a random order by 40 adults aged 18-45 years. Participants ambulated at a comfortable pace for 10 minutes with each device. At baseline and immediately following each trial, a trained operator used diagnostic ultrasonography to capture popliteal vein and artery dimensional and flow characteristics. Results: Significant increases were observed from baseline (0.65 ± 0.23 cm) in venous diameter following walking (0.71 ± 0.21 cm, P = .012) and MKS (0.73 ± 0.21 cm, P = .003). Venous blood flow was also significantly different between conditions ( P = .009) but was only greater following walking (124 ± 79 mL/min) compared to MKS (90 ± 64 mL/min, P = .021). No differences were observed in arterial dimensions between ambulation conditions. Significant increases were found in arterial blood flow from baseline (107 ± 69 mL/min) following walking (184 ± 97 mL/min, P < .001) and HFC (163 ± 86 mL/min, P < .001). Arterial blood flow following walking was greater than AC (132 ± 72 mL/min, P = .016) and MKS (128 ± 74 mL/min, P = .003). Conclusion: We found an average decrease in venous time-averaged mean velocity between walking and use of the MKS, but no such decrease with either HFCs or use of ACs in this healthy experimental cohort. Level of Evidence: Level III, diagnostic comparative study.