Desmopathy of the collateral ligament of the distal interphalangeal joint is a common cause of lameness in the horse and carries a poor prognosis for soundness. Intralesional treatment has been suggested as a way to improve outcome; however, limited reports describe methods for injecting this ligament. This study compares the accuracy of injecting the collateral ligament of the distal interphalangeal joint using magnetic resonance imaging (MRI) versus radiographic guidance. Equine cadaver digit pairs (n=10) were divided by random assignment to injection of the ligament by either technique and assessed using post-injection MRI or gross sections. Images from the proximal, middle, and distal portions of the ligament were blindly evaluated for successful injection. McNemar's test was performed to determine statistical difference between injection techniques. Fisher's exact test was used to evaluate number of injection attempts and injection of the medial or lateral collateral ligament. MRI-guided injection was successful more frequently than radiographic-guided injection on post-injection MRI (24 of 30 versus 9 of 30; p=0.0006) and gross sections (26 of 30 versus 13 of 30; p=0.0008). At each level of the ligament (proximal, middle, and distal), MRI-guided injection resulted in more successful injections than radiographic guidance. Statistical significance occurred at the proximal aspect of the collateral ligament based on post-injection MRI (p=0.0143) and the middle portion of the ligament based on gross sections (p=0.0253). Based on these results, injection guided by standing, low-field MRI should be considered an option for delivering intralesional regenerative therapy to horses with desmopathy of these collateral ligaments.