Actin rings are periodic cytoskeletal structures found in dendrites and neuronal cells that regulate the shape and stability of dendritic spines. They are also known to act as a semi-permeable barrier to membrane diffusion. However, detecting this effect in the diffusion of membrane-bound proteins has been challenging due to their subdiffraction-sized period and the need for simultaneous high localization precision and fast framerates. MINFLUX microscopy, a superresolution technique with unprecedented localization precision, has the potential to overcome these challenges. In this study, we used 3D MINFLUX to track fluorescent quantum dot and nanobody-labelled GPI-anchored proteins on the membrane of stem cell-derived neurons to detect anomalies in their diffusion and highlight the influence of the actin rings on the plasma membrane. Our results demonstrate the high-throughput single particle tracking capabilities of 3D MINFLUX on axon-like structures, leading to the quantification of their diffusion dynamics in 3D. The high localization precision and fast frame rates of MINFLUX microscopy enable researchers to track single fluorescent reporters in living cells and provide new insights into the dynamics of membrane-bound proteins and cytoskeletal structures within neurons. This study highlights the potential of 3D MINFLUX as a powerful single molecule tracking technique with potential applications in basic biology and infection research.