The Moderate Resolution Imaging Spectroradiometer (MODIS) is one of the key instruments for NASA’s Earth Observing System (EOS), currently operating on both the Terra and Aqua satellites. The MODIS is a major advance over the previous generation of sensors in terms of its spectral, spatial, and temporal resolutions. It has 36 spectral bands: 20 reflective solar bands (RSB) with center wavelengths from 0.41 to 2.1 µm and 16 thermal emissive bands (TEB) with center wavelengths from 3.7 to 14.4 µm, making observations at three spatial resolutions: 250 m (bands 1–2), 500 m (bands 3–7), and 1km (bands 8–36). MODIS is a cross-track scanning radiometer with a wide field-of-view, providing a complete global coverage of the Earth in less than 2 days. Both Terra and Aqua MODIS went through extensive pre-launch calibration and characterization at various levels. In orbit, the calibration and characterization tasks are performed using its on-board calibrators (OBCs) that include a solar diffuser (SD) and a solar diffuser stability monitor (SDSM), a v-grooved flat panel blackbody (BB), and a spectro-radiometric calibration assembly (SRCA). In this paper, we present au overview of MODIS calibration and characterization activities, methodologies, and lessons learned from pre-launch characterization and in-orbit operation. Key issues discussed in this paper include in-orbit efforts of monitoring the noise characteristics of the detectors, tracking the solar diffuser and optics degradations, and updating the sensor’s response versus scan angle. The experiences and lessons learned through MODIS have played and will continue to play major roles in the design and characterization of future sensors.