The NASA Curiosity rover Mast Camera (Mastcam) system is a pair of fixed-focal length, multispectral, color CCD imagers mounted ~2 m above the surface on the rover's remote sensing mast, along with associated electronics and an onboard calibration target. The left Mastcam (M-34) has a 34 mm focal length, an instantaneous field of view (IFOV) of 0.22 mrad, and a FOV of 20° × 15° over the full 1648 × 1200 pixel span of its Kodak KAI-2020 CCD. The right Mastcam (M-100) has a 100 mm focal length, an IFOV of 0.074 mrad, and a FOV of 6.8° × 5.1° using the same detector. The cameras are separated by 24.2 cm on the mast, allowing stereo images to be obtained at the resolution of the M-34 camera. Each camera has an eight-position filter wheel, enabling it to take Bayer pattern red, green, and blue (RGB) “true color” images, multispectral images in nine additional bands spanning ~400–1100 nm, and images of the Sun in two colors through neutral density-coated filters. An associated Digital Electronics Assembly provides command and data interfaces to the rover, 8 Gb of image storage per camera, 11 bit to 8 bit companding, JPEG compression, and acquisition of high-definition video. Here we describe the preflight and in-flight calibration of Mastcam images, the ways that they are being archived in the NASA Planetary Data System, and the ways that calibration refinements are being developed as the investigation progresses on Mars. We also provide some examples of data sets and analyses that help to validate the accuracy and precision of the calibration.