During both embryonic development and in adulthood regulation of blood vessel homeostasis is essential for the function of all organs and tissues. As a result, there is a great importance in understanding the mechanisms that regulate vascular development. The ability to control blood vessel growth (i.e. therapeutic vasculogenesis), has the potential to provide specialised treatment for tumour progression, after an ischemic event such as stroke, or aid in wound healing. However, current knowledge of the key players and processes involved is still limited. In order to further elucidate the process involved, multiple publications utilise tube formation assays to demonstrate creation of tubular structures in vitro. New software programmes have also recently been developed to assist with analysis but does this really prove lumen development or are the cells merely aligning into vascular-like structures? Our group has endeavoured to develop a method of 3D visualisation via matrix sectioning and whole mount immunofluorescence of tube structures. This extra step of confirming lumen formation allows for greater confidence that cells are mimicking their true nature in vivo therefore, serving as a critical tool for greater exploration of cell biology and pathologies associated with disease for improved clinical outcomes.