Background: Tissue necrosis, a consequence of inadequate tissue oxygenation, is a common post-operative complication. As current surgical assessments are often limited to visual and tactile feedback, additional techniques that can aid in the interrogation of tissue viability are needed to improve patient outcomes. In this bi-institutional pilot study, the performance of a novel snapshot hyperspectral imaging camera to detect superficial cutaneous oxygen saturation (StO 2 ) was evaluated.Methods: Healthy human volunteers were recruited at two participating centers. Cutaneous StO 2 of the forearm was determined by a snapshot hyperspectral camera on two separate study days during occlusionreperfusion of the brachial artery and after induction of local vasodilation. To calculate the blood StO 2 at each pixel in the multispectral image, spectra were selected, and fitting was performed over wavelengths ranging from 470 to 950 nm. Results: Quantitative detection of physiological changes in cutaneous StO 2 levels was feasible in all sixteen volunteers. A significant (P<0.001) decrease in cutaneous StO 2 levels from 78.3% (SD: 15.3) at baseline to 60.6% (SD: 19.8) at the end of occlusion phase was observed, although StO 2 levels returned to baseline after five minutes. Mean cutaneous StO 2 values were similar in the same subjects on separate study days (Pearson R2: 0.92 and 0.77, respectively) at both centers. Local vasodilation did not yield significant changes in cutaneous StO 2 values.Conclusions: This pilot study demonstrated the feasibility of a snapshot hyperspectral camera for detecting quantitative physiological changes in cutaneous StO 2 in normal human volunteers, and serves as a precursor for further validation in perioperative studies.