In the biomedical field, Near-Infrared Spectroscopy (NIRS) is often used to determine the physiological information by the noninvasive detection of the regional tissue oxygen saturation (rSO 2 ). Because the fatigue degree of body could be obviously demonstrated by the oxygen saturation under different vibration conditions, an effective technology to evaluate the vibration comfortability by rSO 2 is innovatively proposed in this paper. Without losing generality, the vibration comfortability is analyzed by taking the driving position for example. Different from much more factors in previous approaches, only four main body factors, i.e., brain, musculus biceps brachii, erector spinae and biceps femoris, and three main influence factors on vibration comfortability, i.e., acceleration, frequency, and exposure time, are taken into account in our technology. Under different conditions of the three vibration factors, the rSO 2 for the four body factors is detected one by one. Then the relationship among the rSO 2 , subjective feeling, and the vibration parameters can be obtained by the least square method, and the quantitative evaluation model on vibration comfortability is established. It is found experimentally that the rSO 2 trends of musculus biceps brachii, erector spinae and biceps femoris are all greatly increased with the acceleration and frequency, but brain is decreasing obviously. Through our experiment it is verified that this technology can be used in objectively and effectively evaluating under different vibration conditions. This novel technology could provide theoretical support for vibration comfortability assessment and have potential applications in other relative fields.