The characterization of complex mixtures such as cells, cell media, protein biotheraputics, API formulations, and polymeric solutions many times gives more questions than answers. Fluorescence Excitation Emission Matrices (EEMs) have been used for years now to study and even identify multiple components in such complex mixtures. With the development of 2-in-1 UV-visible absorbance and fluorescence spectrometers, the absorbance-transmittance EEM or A-TEEM enabled the acquisition of these matrices to be done more accurately and at higher fluorophore concentration ranges. We present another dimension to the study of complex mixtures with fluorescence spectroscopy using a combination of fluorescence anisotropy (FA) and A-TEEMs that can be used to study mixtures of multiple fluorophores from their local environment in terms of reorientational motion. For a solution of bovine serum albumin (BSA) protein, with multiple dye tracers, the local viscosity is measured in polar and non-polar local environments. Two dyes of varying polarity and solubility are used to track the local viscosity in both polar and non-polar regions of BSA protein in native and denatured forms as well as aggregated and non-aggregated states. Since anisotropy ratio is independent of dye concentration, the matrix maps show clearly any changes in local viscosity of both polar (water-soluble) and non-polar (hydrophobic) regions of the solution. Fluorescence Anisotropy A-TEEMs (FA-ATEEMs) quickly and easily enable the detection of proximate environment changes with respect to local viscosity and motion within protein solutions for multiple regions of the macromolecule.