The low-cost Multispectral Airborne Video System (MAVS), developed using off-the-shelf components, is predominantly being used for agricultural and environmental monitoring applications in Australia. In order to guarantee quality of operational imagery, regular radiometric calibration checks, under both preflight and in-flight environments, are a necessity. While preflight calibration is conducted in the field with relative ease, to conduct in-flight calibration checks during operational missions is difficult, especially when onboard calibration instrumentation is not available. The calibration of MAVS imagery in flight requires a radiative transfer code such as the 6S to simulate the radiation signal at the operational altitude. A comparison of the simulated in-flight calibration equation with the corresponding preflight calibration equation showed a statistically significant agreement for every MAVS band, demonstrating the stability of response of the instrument in the air. This MAVS response stability indicates a potential for calibrating operational airborne imagery using preflight calibration equations. A few simulated in-flight calibration checks, in addition to regular preflight calibrations, could still be carried out during the flight season to verify system radiometric response in the air.