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On June 11, 2000, the first vicarious calibration experiment in support of the Multi-angle Imaging SpectroRadiometer (MISR) was conducted. The purpose of this experiment was to acquire in situ measurements of surface and atmospheric conditions over a bright, uniform area. These data were then used to compute top-of-atmosphere (TOA) radiances, which were correlated with the camera digital number output, to determine the in-flight radiometric response of the on-orbit sensor. The Lunar Lake Playa, Nevada, was the primary target instrumented by the Jet Propulsion Laboratory for this experiment. The airborne MISR simulator (AirMISR) on board a NASA ER-2 acquired simultaneous observations over Lunar Lake. The in situ estimations of top-of-atmosphere radiances and AirMISR measurements at a 20-km altitude were in good agreement with each other and differed by 9% from MISR measurements. The difference has been corrected by adjusting the gain coefficients used in MISR standard product generation. Data acquired simultaneously by other sensors, such as Landsat, the Terra Moderate-Resolution Imaging SpectroRadiometer (MODIS), and the Airborne Visible and Infrared Imaging Spectrometer (AVIRIS), were used to validate this correction. Because of this experiment, MISR radiances are 9% higher than the values based on the on-board calibration. Semiannual field campaigns are planned for the future in order to detect any systematic trends in sensor calibration.