Wideband (38 and 50-260 kHz) target strength of organisms were measured in situ using a towed platform in mesopelagic (200-1000 m depth) layers. Organisms with a gas-inclusion are strong scatterers of sound and acoustically distinct from organisms lacking one. In the mesopelagic zone, some of the fish species and physonect siphonophores have a gas-inclusion. Trawl and multinet biological sampling as well as photographic evidence indicate that in the study area (eastern mid-Atlantic Ocean) the majority of the gas-bearing organisms were fish. Subsequently, using a two-layer viscous-elastic spherical gas backscattering model, physical characteristics such as gas-bladder features and body flesh properties were deduced from the measured backscattering signal of individual gas-bearing fish. Acoustic techniques are non-extractive, can be used for the monitoring and quantification of marine organisms in a time- and cost-effective manner, and suit studies of the mesopelagic zone, which is logistically challenging. Vessel-mounted acoustics, widely used for epipelagic studies, has limitations for mesopelagic studies as the deep organisms are inaccessible to high-frequency (≳100 kHz) acoustic pulses transmitted from the surface due to absorption. Therefore, a towed platform equipped with wideband acoustics has several features that can be utilized for monitoring the mesopelagic dense scattering layers containing mixed species.