An unresolved challenge in osteoarthritis research is characterising the localised intra-tissue mechanical response of articular cartilage. The aim of this study was to explore whether laboratory micro-computed tomography (micro-CT) and digital volume correlation (DVC) permit non-destructive quantification of three-dimensional (3D) strain fields in human articular cartilage. Human articular cartilage specimens were harvested from the knee, mounted into a loading device and imaged in the unloaded and loaded states using a micro-CT scanner. Strain was measured throughout the cartilage volume using the micro-CT image data and DVC analysis. The volumetric DVC-measured strain was within 5% of the known applied strain. Variation in strain distribution between the superficial, middle and deep zones was observed, consistent with the different architecture of the material in these locations. These results indicate DVC method may be suitable for calculating strain in human articular cartilage.