Certain embodiments of the present invention provide methods and systems for calibration of an imaging camera or other image acquisition device. Certain embodiments include characterizing a transformation from a coordinate system of an imager to a coordinate system of a first sensor positioned with respect to the imager using a first off-line calibration. Certain embodiments also include characterizing a transformation from a coordinate system of an imaging camera source to a coordinate system of a second sensor positioned with respect to the imaging camera source using a second off-line calibration. Additionally, certain embodiments include quantifying intrinsic parameters of the imaging camera source based on a transformation from the coordinate system of the imager to the coordinate system of the imaging camera source based on the first and second off-line calibrations and information from the first and second sensors and a transmitter positioned with respect to an object being imaged.