Since its launch in May, 2002, Aqua MODIS has successfully operated for more than 20 years and has continuously generated a wide range of data products that have enabled and supported the remote sensing community and users worldwide for their studies of the Earth’s system by monitoring changes in its key environmental parameters. Although Aqua MODIS, designed with a lifetime requirement of 6 years, is currently operated in its extended mission phase, it continues to make high quality global observations of the Earth’s surface via its 36 spectral bands that cover wavelengths from visible to long-wave infrared. To date, all instrument on-board calibrators (OBC) remain capable of performing their design functions, providing various calibration data sets to help monitor on-orbit changes in sensor responses and performance characteristics. In addition to the OBC, regularly scheduled lunar observations and select Earth-view targets are used extensively to support sensor on-orbit calibration, especially for the calibration of the visible channels (or bands). We provide an overview of Aqua MODIS on-orbit calibration activities and methodologies for both reflective solar bands and thermal emissive bands, illustrate its on-orbit performance over the past 20 years using examples derived from OBC measurements, lunar observations, and Earth-view response trends, and describe various calibration improvements made over its entire mission. We focus on key issues identified since launch, such as solar diffuser degradation, electronic crosstalk, and on-orbit changes in sensor response versus scan-angle, along with approaches and strategies developed to mitigate their impact on sensor calibration quality. Also discussed in this paper are some of the key calibration enhancements incorporated recently in the Collection 6.1 and the upcoming Collection 7 Level-1B algorithms.