Retrieval of aerosol loading in vertical atmospheric columns is a common product of satellite and ground instruments that measure spectral extinction of solar radiation throughout the entire atmosphere. Here we study ground hyperspectral imaging of artificial light sources as a complementary method for retrieving fine aerosol concentrations along quazi-horizontal ambient open paths. Previously, we reported hyperspectral measurements of the aerosol optical thickness in the 500–900 nm range over urban-scale distances (180 m to 4 km), measuring the extinction of radiation emitted from a halogen source. Here we confirm in a laboratory-setup the basic premise that different accumulation-size aerosols generate distinct hyperspectral signatures in this spectral range. Measured hyperspectral attenuation signatures of fine aerosols were comparable to calculated Mie scattering signatures, suggesting that modal aerosol concentrations can be retrieved. A genetic algorithm was adapted to estimate the aerosol modal concentrations from its hyperspectral extinction signature. Retrievals of aerosol concentrations from measured and synthetic hyperspectral signatures indicated a robust algorithm, with an expected retrieval error of 0.2–22% for typical ambient concentrations along an urban-scale open path. The retrieval accuracy was found to depend on the relative aerosol modal concentrations, especially when there is a substantial overlap between the modal spectral signatures.