Blood vessel staining in the myocardium for 3D visualization down to the smallest capillaries

Blood vessels formed after medical interventions such as radiofrequency treatment have to be visualized down to the capillary level with diameters of about 5 μm to validate neo-vascularization. Synchrotron radiation-based micro-computed tomography (SRμCT) provides the necessary spatial resolution. Since both the vessels and the surrounding tissue mainly consist of water the difference in absorption is extremely weak. Therefore, it is necessary to search for appropriate contrast agents and to develop suitable staining protocols, which finally allow segmenting the vessel tree. Among the contrast agents used in medicine lyophilic salts with a mean particle diameter of 1.5 μm such as CaSO4, SrSO4 and BaSO4 are most appropriate to stain the vessels. The combination of these salts with a commercially available embedding kit (JB-4, Polysciences Inc.) allows tissue fixation and long-term storage in solid state. Intensity-based segmentation algorithms enable the vessel tree extraction in selected parts of the stained myocardium using the SRμCT data.