Soft tissue injury screening faces two main diagnostic challenges. One is the perceptual bias of the athlete in terms of referred pain and second injury assessment tools are not only in need of highly specialized personal but they are also financially demanding. Since ankle sprains is one of the leading soft tissue injuries, the current study was set to examine the suitability of functional infrared thermal imaging (fITI) in evaluating physiological alteration on the ankle as a result of exercise or injury. The current study consists of a case report of a patient with an ankle sprain and the behavior of temperature after a series of physiotherapy sessions. Moreover to strengthen the communication of the report, results from a healthy population sample were added to draw a deeper understanding on physiological temperature manifestations on soft tissue. Twenty participants underwent a 30-minute treadmill run with pictures of their ankles being taken during rest and after exercise. In addition the case of a patient is reported that has suffered an ankle sprain followed for a period of over a month. It was observed that the temperature of the ankles of participants that underwent physical exercise rose on average by 2.4 °C after taking into account both the medial and lateral sides of the both ankles. In addition the patient’s left ankle appeared to be 2.5 °C above the temperature of the non-affected right ankle. This phenomenon of unilateral hyperthermia of the left injured side seems to start to dissipate by the 21st day following the injury, completely resolving by the 42nd day achieving bilateral isothermia in both ankles. Thermal imaging provides a reliable tool for the screening of soft tissue strain and injury. The current study further expands the literature on soft tissue screening with the use of thermal imaging, adding a quantifiable way for assessing the extend of tissue damage. The implemented method of analyses offers a suggested simple way not only in visualizing trauma but also physical strain. Nevertheless further investigations with a variety in the severity of ankle sprains need to be applied in order for thermal imaging to be used as a first line tool for the assessment and recovery of ankle sprains.