Atmospheric nitrous oxide (N2O) is, after carbon dioxide and methane, the third most important long-lived anthropogenic greenhouse gas in terms of radiative forcing. Since preindustrial times a rising trend in the global N2O concentrations is observed. Anthropogenic emissions of N2O, mainly from agricultural activity, contribute considerably to this trend. Sparse observational constraints have made it difficult to quantify these emissions. The few studies on top-down approaches in the U.S. that exist are mainly based on Lagrangian models and ground-based measurements. They all propose a significant underestimation of anthropogenic N2O emission sources in established inventories, such as the Emissions Database for Global Atmospheric Research (EDGAR). In this study we quantify anthropogenic N2O emissions in the Midwest of the U.S., an area of high agricultural activity. In the course of the Atmospheric Carbon and Transport – America (ACT-America) campaign spanning from summer 2016 to summer 2019, an extensive dataset over four seasons has been collected including in-situ N2O aircraft based measurements in the lower and middle troposphere onboard NASA’s C-130 and B-200 aircraft. During fall 2017 and summer 2019 we conducted measurements onboard the NASA-C130 with a Quantum-Cascade-Laser-Spectrometer (QCLS) and on both aircraft over the whole campaign flask measurements (NOAA) were collected. More than 300 joint flight hours were conducted and more than 500 flask samples were collected over the U.S. Midwest. The QCLS system collected continuous N2O data for approximately 60 flight hours in this region. The Eulerian Weather Research and Forecasting model with chemistry enabled (WRF-Chem) is being used to quantify regional agricultural N2O emissions using the spatial characteristics of these atmospheric N2O mole fraction observations. The numerical simulations enable potential surface emission distributions to be compared to our airborne measurements, and source estimates can be adjusted to minimize the differences, thus quantifying N2O sources. These results are then compared to emission rates in the EDGAR inventory.