Colloidal glasses (CGs) made of polymer (polymethylmethacrylate) nanoparticles are promising metamaterials for light and sound manipulation, but fabrication imperfections and fragility can limit their functionality and applications. Here, the vibrational mechanical modes of nanoparticles are probed to evaluate the nanomechanical and morphological properties of various CGs architectures. Utilizing the scanning micro‐Brillouin light scattering (µ‐BLS), the effective elastic constants and nanoparticles' sizes is determined as a function of position in a remote and non‐destructive manner. This method is applied to CG mesostructures with different spatial distributions of their particle size and degree of order. These include CGs with single‐sized systems, binary mixtures, bilayer structures, continuous gradient structures, and gradient mixtures. The microenvironments govern the local mechanical properties and highlight how the granular mesostructure can be used to develop durable functional polymer colloids. A size effect is revealed on the effective elastic constant, with the smallest particles and ordered assemblies forming robust structures, and classify the various types of mesoscale order in terms of their mechanical stiffness. The work establishes scanning µ‐BLS as a tool for mapping elasticity, particle size, and local structure in complex nanostructures.