OBJECTIVE To investigate the distribution and nerve staining of two volumes of lidocaine-dye solution after ultrasound-guided erector spinae plane (ESP) injections in canine cadavers. METHODS Experimental cadaveric study. METHODS A total of nine canine cadavers. METHODS ESP injections were performed between the longissimus thoracis muscle and the dorsolateral edge of the ninth thoracic transverse process. Two cadavers were transversally cryosectioned after unilateral ESP injections [0.6 mL kg-1; high volume (HV)]. In seven cadavers, bilateral ESP injections with HV or low volume (0.3 mL kg-1; LV) were performed. Gadodiamide was added to the injectate for two cadavers and magnetic resonance imaging (MRI) was performed pre- and post-injection. Injectate distribution and nerve staining of the branches of the spinal nerves were recorded after gross anatomical dissection. The thoracic paravertebral and epidural spaces were examined for dye solution. RESULTS Cryosections, MRI and gross dissections showed that the injectate spread dorsally to the transverse processes, over the ventromedial aspect of the longissimus thoracis muscle where the medial and lateral branches of the dorsal branches of the spinal nerves are located. LV and HV stained a median (range) of 4 (2-7) and 4 (3-8) medial branches, respectively (p = 0.52). LV and HV stained 4 (2-5) and 5 (4-7) lateral branches (p = 0.26), respectively. Ventral branches were not stained, and dye was not identified in the epidural or paravertebral spaces. CONCLUSIONS and clinical relevance Medial and lateral branches were consistently stained over several spinal segments. The number of nerves stained was not different with HV or LV, and the ventral branches of the spinal nerves were not stained in any cadaver. ESP block may find a clinical application to desensitize structures innervated by the medial and lateral branches of the dorsal branches of the thoracic spinal nerves.