Powered lower limb prostheses have potential to improve the quality of life of individuals with amputations by enabling all daily activities. However, seamless ambulation mode recognition is necessary to achieve this goal and is not yet a clinical reality. Current intent recognition systems use mechanical and EMG sensors to estimate prosthesis and user status. We propose to complement these systems by integrating information about the environment obtained through the depth sensing. This paper presents the design, characterization, and the early validation of a novel stair segmentation system based on Microsoft Kinect. Static and dynamic tests were performed. A first experiment showed how the resolution of the depth camera affects the speed and the accuracy of segmentation. A second test proved the robustness of the algorithm to different staircases. Finally, we performed an online walking test with the stair segmentation and related measures recorded online at >5 frames/s. Experimental results show that the proposed algorithm allows for an accurate estimate of distance, angle of intersection, number of steps, stair height, and stair depth for a set of stairs in the environment. The online test produced an estimate of whether the individual was approaching stairs in real time with approximately 98.8% accuracy.