This prospective anatomical study aimed to establish an ultrasound-guided technique to the quadratus lumborum (QL) plane in sheep cadavers. Thirteen cadavers, weighing less than 117 kg, were included. In phase 1, one cadaver underwent dissection and two cadavers underwent 3D computed tomographic reconstruction for anatomical evaluation of the thoracolumbar region. In phase 2, two cadavers were used to compare two ultrasound techniques to the QL plane: lateral to the QL muscle with a transversal approach (LQL) and transmuscular between QL and psoas muscles with a longitudinal approach (TQL). For LQL, the reference was the first lumbar transverse process, whereas for TQL, it was the intertransverse region between the first and second lumbar vertebrae. The needle was advanced in-plane towards the specific target for each technique and a total of four injections were performed using 0.4 ml kg−1 of a dye-lidocaine solution. In phase 3, 10 cadavers received bilateral LQL injections ( n = 20). All cadavers were then dissected to evaluate spread of dye. In phase 2, following LQL injections, no dye was observed in undesired locations; however, the dye was noted in the retroperitoneal space (1/2) after TQL injections. In phase 3, the 13th thoracic, first, second, third lumbar nerves, and sympathetic trunk segments were stained in 80%, 95%, 100%, 45% and 35% of the injections, respectively. In conclusion, the LQL technique was feasible, allowing staining of the spinal nerves innervating the cranial abdomen in sheep cadavers. Further studies in live animals are warranted.