Background: Cardiovascular disease (CVD) is associated with impairments in microvascular responsiveness. Therefore, reliably assessing microvascular function is clinically relevant. Thus, this study aimed to examine the reliability of the near-infrared spectroscopy (NIRS)-derived oxygen saturation (StO(2)) reperfusion slope, a measure of microvascular responsiveness, to four different vascular occlusion tests (VOT) of different durations in young and older participants. Methods: Eight healthy young (29 +/- 5 yr) and seven older (67 +/- 4 yr) men participated in four NIRS combined with VOT (NIRS-VOT; 30 s, 1, 3, and 5 min) in the leg microvasculature on two visits separated by 1-2 weeks. Vascular responsiveness was determined by the StO(2) reperfusion slope. The coefficient of variation (CV), repeatability, reliability (ICC), and the limits of agreement (LOA) were calculated for the NIRS-derived reperfusion slopes for each occlusion duration and visit. Results: CV for the StO(2) reperfusion slope following 30 s, 1, 3 and 5 min of occlusion were 33 +/- 29%, 19 +/- 21%, 14 +/- 12%, and 12 +/- 10%, respectively. Repeatability values following 30 s, 1, 3 and 5 min occlusions were 20%, 1%, 4% and 21%, respectively. The ICC for the St0 2 reperfusion slopes for each occlusion duration were 0.29, 0.42, 0.84, and 0.88 following 30 s, 1, 3 and 5 min of occlusion, respectively. LOA values between visit 1 and 2 for occlusions were not different from zero. There were no age-related differences for all variables of the study. Conclusion: NIBS-derived StO(2) reperfusion slope, has good reliability across a range of occlusion durations with the strongest reliability during longer occlusion durations.