Analysis of visible remote sensing data research requires removing atmospheric effects by conversion from radiance to at-surface reflectance. This conversion can be achieved through theoretical radiative transfer models, which yield good results when well-constrained by field observations, although these measurements are often lacking. Additionally, radiative transfer models often perform poorly in marine or lacustrine settings or when complex air masses with variable aerosols are present. The empirical line method (ELM) measures reference targets of known reflectance in the scene. ELM methods require minimal environmental observations and are conceptually simple. However, calibration coefficients are unique to the image containing the reflectance reference. Here we compare the conversion of hyperspectral radiance observations obtained with the NASA Glenn Research Center Hyperspectral Imager to at-surface reflectance factor using two reflectance reference targets. The first target employs spherical convex mirrors, deployed on the water surface to reflect ambient direct solar and hemispherical sky irradiance to the sensor. We calculate the mirror gain using near concurrent at-sensor reflectance, integrated mirror radiance, and in situ water reflectance. The second target is the Lambertian-like blacktop surface at Maumee Bay State Park, Oregon, OH, where reflectance was measured concurrently by a downward looking, spectroradiometer on the ground, the aerial hyperspectral imager and an upward looking spectroradiometer on the aircraft. These methods allows us to produce an independently calibrated at-surface water reflectance spectrum, when atmospheric conditions are consistent. We compare the mirror and blacktop-corrected spectra to the in situ water reflectance, and find good agreement between methods. The blacktop method can be applied to all scenes, while the mirror calibration method, based on direct observation of the light illuminating the scene validates the results. The two methods are complementary and a powerful evaluation of the quality of atmospheric correction over extended areas. We decompose the resulting spectra using varimax-rotated, principal component analysis, yielding information about the underlying color producing agents that contribute to the Ortiz et al.Vicarious Hyperspectral Reflectance Calibration Methods observed reflectance factor scene, identifying several spectrally and spatially distinct mixtures of algae, cyanobacteria, illite, haematite, and goethite. These results have implications for future hyperspectral remote sensing missions, such as PACE, HyspIRI, and GeoCAPE.