In this study, nanoindentation was utilized to measure the local, three-dimensional properties of Kevlar 49 and Kevlar KM2 on the length scales of the fiber microstructure. First, atomic force microscopy-based methods were used to explore the extent of property changes with respect to radial position in the fibers’ axial and hoop planes. From these measurements, no significant change in response was found for Kevlar 49 fibers, consistent with transverse isotropy. However, a reduced stiffness “shell” region (up to ∼300–350 nm thick) was observed for KM2 fibers. Instrumented indentation was then used to evaluate fiber response with respect to orientation and contact size and establish a critical contact size above which the response is independent of indenter size (i.e., “homogeneous” behavior). A previously proposed analytical method for indentation of a transversely isotropic material was used to estimate the local material properties of the Kevlar fibers from the measured homogeneous response.