This experimental work focused on the sensor selection for the development of a needle-like instrument to treat small isolated cartilage defects with hydrogels. The aim was to identify the most accurate and sensitive imaging method to determine the location and size of defects compared to a gold standard (µCT). Only intravascular ultrasound imaging (IVUS) vs. optical coherent tomography (OCT) were looked at, as they fulfilled the criteria for integration in the needle design. An in-vitro study was conducted on six human cadaveric tali that were dissected and submerged in saline. To simulate the natural appearance of cartilage defects, three types of defects were created via a standardised protocol: osteochondral defects (OCD), chondral defects (CD) and cartilage surface fibrillation (CSF), all sized between 0.1 and 3 mm in diameter. The detection rate by two observers for all diameters of OCD were 80, 92 and 100% with IVUS, OCT and µCT, for CD these were 60, 83 and 97%, and for CSF 0, 29 and 24%. Both IVUS and OCT can detect the presence of OCD and CD accurately if they are larger than 2 mm in diameter, and OCT can detect fibrillated cartilage defects larger than 3 mm in diameter. A significant difference between OCT–µCT and IVUS–µCT was found for the diameter error (p = 0.004) and insertion depth error (p = 0.002), indicating that OCT gives values closer to reference µCT. The OCT imaging technique is more sensitive to various types and sizes of defects and has a smaller diameter, and is therefore preferred for the intended application.