The term "sagittal stratum" was coined by Heinrich Sachs in 1892 to define a parasagittally oriented white matter layer at the temporo-occipital cortex. Although this term has been widely used for more than 100 years, the description, classification, borders, and involved fibers of the structure vary among authors and remain imprecise. Through fiber microdissection and tractography, the authors aimed to define the sagittal stratum and resolve the uncertainty by revealing the relationship of this structure to other cerebral white matter pathways and the orientation of fibers in it. Twenty postmortem human cerebral hemispheres were prepared according to Klingler's method. Fiber dissections were performed under a surgical microscope and with microsurgical techniques. The results of dissection at each step were photographed with 2D and 3D imaging techniques, and 3D photogrammetry techniques were used to create a 360° model. Diffusion tensor imaging and 7T high-resolution MRI were used to confirm the findings. This study revisited the 3D organization of white matter tracts in the sagittal stratum through fiber microdissection and tractography. The microneuroanatomical structure of the sagittal stratum and its special organization with fibers from all three fiber systems are demonstrated. The authors' findings revealed that the sagittal stratum has two layers consisting of four different fiber tracts. Its external layer consists of a long association fiber and a commissural fiber, while its internal layer consists of intertwined projection fibers, including temporo-parieto-occipitopontine fibers and the posterior thalamic peduncle. Detailed microdissection also showed the location of the posterior thalamic peduncle in the most medial site of all posterior hemispheric projection fibers. The structure of the sagittal stratum is distinctive in that it contains all three main fiber systems: association, commissural, and projection. Because of its expansive location in the temporal and occipital lobes, it can be damaged by most neurosurgical pathologies and procedures. The authors emphasize the significance of preserving the sagittal stratum during surgical interventions while also challenging the notion of a "silent" brain, suggesting that the current inability to fully comprehend cerebral function contributes to this misconception. Detailed knowledge of the complex white matter anatomy of the sagittal stratum can guide neurosurgeons in surgical planning and the selection of appropriate surgical approaches with intraoperative orientation for safe surgery and less comorbidity.