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This study reports on the utilization of microwave modeling, together with ground truth, and L-band (1.4-GHz) brightness temperatures to investigate the passive microwave characteristics of a conifer forest floor. The microwave data were acquired over a natural Virginia Pine forest in Maryland by a ground-based microwave active/passive instrument system in 2008/2009. Ground measurements of the tree biophysical parameters and forest floor characteristics were obtained during the field campaign. The test site consisted of medium-sized evergreen conifers with an average height of 12 m and average diameters at breast height of 12.6 cm. The site is a typical pine forest site in that there is a surface layer of loose debris/needles and an organic transition layer above the mineral soil. In an effort to characterize and model the impact of the surface litter layer, an experiment was conducted on a day with wet soil conditions, which involved removal of the surface litter layer from one half of the test site while keeping the other half undisturbed. The observations showed detectable decrease in emissivity for both polarizations after the surface litter layer was removed. A first-order radiative transfer model of the forest stands including the multilayer nature of the forest floor in conjunction with the ground truth data are used to compute forest emission. The model calculations reproduced the major features of the experimental data over the entire duration, which included the effects of surface litter and ground moisture content on overall emission. Both theory and experimental results confirm that the litter layer increases the observed canopy brightness temperature and obscure the soil emission.