Background Optical coherence tomography (OCT) is a high-resolution sub-surface imaging modality using near-infrared light to provide accurate and high contrast intra-vascular images. This enables accurate assessment of diseased arteries before and after intravascular intervention. This study was designed to corroborate diagnostic imaging equivalence between the Ocelot and the Dragonfly OCT systems with regards to the intravascular features that are most important in clinical management of patients with atherosclerotic vascular disease. These intravascular features were then corroborated in vivo during treatment of peripheral arterial disease (PAD) pathology using the Ocelot catheter. Methods In order to compare the diagnostic information obtained by Ocelot (Avinger Inc., Redwood City, CA) and Dragonfly (St. Jude Medical, Minneapolis, MN) OCT systems, we utilized ex-vivo preparations of arterial segments. Ocelot and Dragonfly catheters were inserted into identical cadaveric femoral peripheral arteries for image acquisition and interpretation. Three independent physician interpreters assessed the images to establish accuracy and sensitivity of the diagnostic information. Histologic evaluation of the corresponding arterial segments provided the gold standard for image interpretation. In vivo clinical images were obtained during therapeutic interventions that included crossing of peripheral chronic total occlusions (CTOs) using the Ocelot catheter. Results Strong concordance was demonstrated when matching image characteristics between both OCT systems and histology. The Dragonfly and Ocelot system’s vessel features were interpreted with high sensitivity (91.1–100 %) and specificity (86.7–100 %). Inter-observer concordance was documented with excellent correlation across all vessel features. The clinical benefit that the Ocelot OCT system provided was demonstrated by comparable procedural images acquired at the point of therapy. Conclusions The study demonstrates equivalence of image acquisition and consistent physician interpretation of images acquired by the Ocelot and the Dragonfly OCT systems in-spite of distinct image processing algorithms and catheter configurations. This represents a dramatic shift away from both fluoroscopic imaging and diagnostic-only OCT imaging during peripheral arterial intervention towards therapeutic devices that incorporate real time diagnostic OCT imaging. In the clinical practice, these diagnostic capabilities have translated to best-in-class safety and efficacy for CTO crossing using the Ocelot catheter.