In cancer research, well characterized small animal models of human cancer, such as transgenic mice, have greatly accelerated the pace of development of cancer treatments. The goal of the Small Animal Radiation Research Platform (SARRP) is to make those same models available for the development and evaluation of novel radiation therapies. SARRP can deliver high resolution, sub millimeter, optimally planned conformal radiation with on-board cone-beam CT (CBCT) guidance. This system requires accurate calibration of the x-ray beam for both imaging and radiation treatment. In this paper, we present a novel technique using an x-ray camera for calibration of the treatment beam. This technique does not require precise positioning or calibration of the x-ray camera.