AbstractRemote sensing with uncrewed aerial vehicles (UAVs) is increasingly being used in agriculture to provide data on the physical characteristics of plants under field conditions. Data accuracy is critical for decision making with a high degree of confidence. In this work, we compared two multispectral camera calibration methods for image data collected with a UAV: (1) an autoexposure method that relies on a single calibration panel and a post hoc calibration, and (2) a fixed‐exposure system that uses three in‐field gray calibration panels using the empirical line calibration method. Both methods were compared to reflectance data from (a) four ground calibration targets measured with a spectroradiometer and (b) a single manned aircraft image calibrated with commercial calibration tarps. In a band‐by‐band comparison, the autoexposure method produced almost twice as much radiometric error on average compared with fixed exposure. Because remote sensing data are commonly converted to spectral indices, the calibration methods were also evaluated by calculating the visible atmospherically resistant index (VARI) and comparing the resulting data to the manned aircraft image. Similarly, the autoexposure method in this case produced twice the error of the fixed‐exposure method. The effect of the error was considered in a production agriculture context by simulating a remote sensing‐based prescription map for pesticide application in a cotton (Gossypium) field and calculating the number of mislabeled management zones. The simulation showed that the autoexposure method would be more costly to the farm because of its higher error, roughly $8.00/ha based on the assumptions made.