Near-infrared diffuse correlation spectroscopy (NIR-DCS) is an emerging technology for simultaneous measurement of skeletal muscle microvascular oxygen delivery and utilization during exercise. The extent to which NIR-DCS can track acute changes in oxygen delivery and utilization has not yet been fully established. To address this knowledge gap, 14 healthy men performed rhythmic handgrip exercise at 30% maximal voluntary contraction, with and without isolated brachial artery compression, designed to acutely reduce convective oxygen delivery to the exercising muscle. Radial artery blood flow (Duplex Ultrasound) and NIR-DCS derived variables [blood flow index (BFI), tissue oxygen saturation ([Formula: see text]), and metabolic rate of oxygen ([Formula: see text])] were simultaneously measured. During exercise, both radial artery blood flow (+51.6 ± 20.3 mL/min) and DCS-derived BFI (+155.0 ± 82.2%) increased significantly ( P < 0.001), whereas [Formula: see text] decreased −7.9 ± 6.2% ( P = 0.002) from rest. Brachial artery compression during exercise caused a significant reduction in both radial artery blood flow (−32.0 ± 19.5 mL/min, P = 0.001) and DCS-derived BFI (−57.3 ± 51.1%, P = 0.01) and a further reduction of [Formula: see text] (−5.6 ± 3.8%, P = 0.001) compared with exercise without compression. [Formula: see text] was not significantly reduced during arterial compression ( P = 0.83) due to compensatory reductions in [Formula: see text], driven by increases in deoxyhemoglobin/myoglobin (+7.1 ± 6.1 μM, P = 0.01; an index of oxygen extraction). Together, these proof-of-concept data help to further validate NIR-DCS as an effective tool to assess the determinants of skeletal muscle oxygen consumption at the level of the microvasculature during exercise.