Background <p>The superior longitudinal fasciculus/arcuate fasciculus (SLF/AF) connects the temporal and frontal lobes through the parietal region. Although well-studied, the SLF/AF arterial supply and 3D-relationship with the middle cerebral artery (MCA) have not been reported. We examined this using cadaveric fiber dissection and tractography to provide neurosurgical-relevant 3D information.</p> Methods <p>Ten cadaveric hemispheres were removed, vascular tissues cleaned, and fiber dissection performed. In another 10 hemispheres, artery-preserving dissections were carried out in situ. Diffusion Tensor Imaging tractography and Magnetic Resonance Angiography (MRA) of 10 healthy individuals were 3D reconstructed with consistent nomenclature.</p> Results <p>Fiber dissections revealed the superior longitudinal complex (SLF-C), the superior longitudinal fasciculus II (SLF-II), the superior longitudinal fasciculus III (SLF-III), and the arcuate fasciculus (AF) fiber tracts in vascular and meningeal cleared hemispheres. These fiber tracts were also revealed in five heads with intact MCA arterial structures from the MCA bifurcation to the medullary branches. This provided visualization of the SLF/AF and its 3D relationship with the MCA. SLF-C, SLF-II, SLF-III, and AF were shown individually and together with tractography of healthy individuals. The 3D relationship between these fiber tracts, the MCA and its branches was demonstrated by overlaying MRA vascular segmentations onto tractography images.</p> Conclusion <p>This is the first report showing the SLF/AF and MCA relationship as a whole, from its cortical to its medullary branches, by overlaying tractography with MRA reconstructions. This 3D analysis provides valuable insights into their spatial organization to enhance surgical practice and understand the vascular associations of the SLF/AF.</p>