A precise anatomical understanding of the thoracic paravertebral space (TPVS) is essential to understanding how an injection outside this space can result in paravertebral spread. Therefore, we aimed to clarify the three-dimensional (3D) structures of the TPVS and adjacent tissues using micro-CT, and investigate the potential routes for nerve blockade in this area.Eleven embalmed cadavers were used in this study. Micro-CT images of the TPVS were acquired after phosphotungstic acid preparation at the mid-thoracic region. The TPVS was examined meticulously based on its 3D topography.Micro-CT images clearly showed the serial topography of the TPVS and its adjacent spaces. First, the TPVS was a very narrow space with the posterior intercostal vessels very close to the pleura. Second, the superior costotransverse ligament (SCTL) incompletely formed the posterior wall of the TPVS between the internal intercostal membrane and vertebral body. Third, the retro-SCTL space broadly communicated with the TPVS via slits, costotransverse space, intervertebral foramen, and erector spinae compartment. Fourth, the costotransverse space was intersegmentally connected to the adjacent retro-SCTL space.A non-destructive, multi-sectional approach using 3D micro-CT more comprehensively demonstrated the real topography of the intricate TPVS than previous cadaver studies. The posterior boundary and connectivity of the TPVS provides an anatomical rationale for the notion that paravertebral spread can be achieved with an injection outside this space.