The murine model of hindlimb ischemia (HLI) is among the most popular useful models to study vascular disease. However, from a physiological perspective, the available information regarding the murine vascular function regulation is practically inexistent. Our objective was to characterize the bilateral vascular response to a normobaric oxygen breathing (NOB) protocol in two groups (i) a control group, consisting of 16 healthy male C57BL/6 mice (8-27 weeks old), and an (ii) HLI model group, consisting of 9 male C57BL/6 mice (16 weeks old). All procedures involving animal experimentation were ethically supervised. Ischemia was surgically induced on the left limb under isoflurane anesthesia, while the right limb remained as the control. The NOB protocol included a 10 min stabilization period, a 10 min breathing a saturated oxygen atmosphere, and further 10 min for recovery. Mice were maintained under ketamine-xylazine anesthesia. The NOB procedure was applied once to the control group, and on day 0 (surgery) and 4, 6, 9, 12, 15 and 21 post-surgery days in the HLI group. Blood flow was recorded in both paws by LDF. On the HLI group perfusion was recorded with both LDF and LDI. Spectral characterization of the LDF signal was performed with Fourier and wavelet transforms. Nonparametric statistics were applied (p<0.05). Three main vascular responses were detected on the control group with NOB – bilateral perfusion decrease, bilateral perfusion increase and mixed response. For the HLI group no differences in perfusion between paws were found in day 0. In the HLI group, after surgery, the control paw consistently responded to NOB with a perfusion decrease, while for the ischemic paw, an increase in perfusion was consistently observed in all days. In these animals LDF and LDI perfusion signals were positively correlated in all recovery days in both paws. This approach also allowed the spectral characterization of the murine LDF signal and respective perfusion-regulating components frequency ranges (heart, respiration, myogenic, sympathetic and endothelial) which reinforces the potential usefulness of this animal model to look deeper into vascular physiology.