Bridging Histology and Tractography: First In Vivo Visualization of Short-Range Prefrontal Connections Informed by...

Decades of histological research in non-human primates have revealed a dense web of short-range connections underpinning prefrontal cortex (PFC) function. However, translating this anatomical ground-truth to the living human brain has been a major challenge, leaving our understanding of the PFC's intrinsic wiring incomplete. These short-range fibers are difficult to resolve with non-invasive methods like diffusion tractography, which are often hampered by false positives. Here, we provide the first systematic in vivo visualization of these pathways in the human brain. By informing high-resolution probabilistic tractography with established tract-tracing findings, we mapped 91 histologically-defined short-range connections within and between five major PFC subdivisions in 1003 individuals (547 F, 456 M). Our anatomically-informed approach successfully reconstructed these intricate connections with high precision (> 80%) and accuracy (> 70%) relative to histological findings. The resulting tracts not only captured broad organizational principles but also replicated fine-grained patterns previously only seen in invasive studies. Furthermore, these connections showed high test-retest reliability within individuals alongside significant variability between them, highlighting a stable yet unique anatomical fingerprint. Ultimately, this study shows how linking histology to tractography provides a powerful framework to advance our understanding of the human connectome and opens avenues to investigate local circuitry that underpins cognition and disease.