Systems and methods are disclosed for calibrating an imaging system comprising a processing unit communicatively coupled to one or more imaging devices. The imaging system may also include a display. In an embodiment, a sequence of display features are displayed on a display, and imaged by the imaging device or devices. Spatial data is compiled related to the display features and the corresponding image features. In an embodiment, the display, imaging device or device, or both may be repositioned and another sequence of display features may be presented and the spatial data compiled. Using optimization and calibration techniques, the compiled spatial data may be used to obtain one or more internal imaging device parameters, one or more external parameters, or both.