MRI-based models enable non-invasive characterization of brain microstructure, e.g. the effective axon radius ($$$r_{\text{eff}}$$$). Often, these models were validated by small-field-of-view microscopy (sFoVM) images (~10³ axons). As $$$r_{\text{eff}}$$$ is dominated by large, sparsely occurring axons, sFoVM-based estimations may miss large axons and underestimate $$$r_{\text{eff}}$$$. We employed an in-house developed pipeline to estimate $$$r_{\text{eff}}$$$ on large-scale light microscopy (lsLM) sections with similar spatial extent as the voxel sizes in human MRI systems. Taking lsLM as a baseline, we showed that sFoVM is an unsuitable reference for $$$r_{\text{eff}}$$$ and verified the potential of $$$r_{\text{eff}}$$$ to capture relevant spatial, anatomical variation.
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