OBJECTIVES: This proof-of-concept study aimed to assess the feasibility of Multispectral Optoacoustic Tomography (MSOT) in evaluating changes in oxygenated hemoglobin (HbO2) levels in muscles of the lower limb before and after lower extremity revascularization (LER). METHODS: In 26 patients, HbO2 levels were assessed before and after LER, with follow-up assessing symptom control and patency for up to six months. RESULTS: A significant difference in HbO2 levels was observed between pre- and post-LER in the muscles of the lower limb. In 10 patients, HbO2 levels did not increase following LER, and at the 6-month follow-up, 2 of these patients required target lesion revascularization (TLR) due to restenosis of ≥ 50 % stenosis. In contrast, 16 patients demonstrated increased HbO2 levels post-LER, with no patients requiring TLR at 6-months. CONCLUSION: This study demonstrates the potential of MSOT to detect changes in tissue perfusion following LER, highlighting its promise as a novel imaging modality for guiding treatment strategies.