One Sentence Summary: This is the first detailed analysis of the segment-specific dorsal and ventral spinal roots spatial orientation measured and correlated to the anatomical landmarks of the spinal cord and vertebral column for human.Abstract: An understanding of spinal cord functional neuroanatomy is essential for diagnosis and treatment of multiple disorders including, chronic pain, movement disorders, and spinal cord injury. Till now, no information is available on segment-specific spinal roots orientation in humans. In this study we collected neuroanatomical measurements of the dorsal and ventral roots from C2-L5, as well as spinal cord and vertebral bone measurements from adult cadavers. Spatial orientation of dorsal and ventral roots were measured and correlated to the anatomical landmarks of the spinal cord and vertebral column. The results show less variability in rostral root angles compared to the caudal angles across all segments. Dorsal and ventral rootlets were oriented mostly perpendicular to the spinal cord at the cervical level and demonstrate more parallel orientation at the thoracic and lumbar segments. The number of rootlets was the highest in dorsal cervical and lumbar segments. Spinal cord transverse diameter and size of the dorsal columns were largest at cervical and lumbar segments. The strongest correlation was found between the length of intervertebral foramen to rostral rootlet and vertebral bone length. These results could be used to locate spinal roots and spinal cord landmarks based on bone marks on CT or X-rays. These results also provide background for future correlations between anatomy of spinal cord and spinal column structures that could improve stereotactic surgical procedures and electrode positioning for spinal cord neuromodulation.Intervertebral foramen to rostral rootlet distance: Similar to previous measurement, intervertebral foramen to rostral rootlet distance was gradually increased to the greatest values at lumbar segments (9.59 ± 2.55 cm) (p<0.001) (Fig. 1G). This distance at C2 segment was 1.55 ± 0.42 cm and it did not change significantly throughout cervical segments till C7, where C7 (2.01 ± 0.08 cm) and C8 (2.27 ± 0.22 cm) were significantly greater compare to C5. Lower cervical and upper thoracic segments did not vary significantly till T3, where there it increased (4.05 ± 0.63 cm). No other major variations were observed across the rest of thoracic segments except for T12 (5.68 ± 0.92 cm) that was significantly longer compare to T1-T10. T12, L1, and L2 segments did not vary, but L3 (10.27 ± 2.23 cm) showed significant increase compared to L1. No major variations were observed for the rest of lumbar segments, although at L5 (12.66 ± 1.50 cm) it was significantly longer than at L2 (Fig. 1I, purple line).