3-D Visualisation of Chicken Embryo Vasculature

Shear stress responsive genes in the vessel walls of§the developing chicken embryo have a large impact on§the development of the vasculature. To investigate§the effect of altered blood flow on vessel formation,§blood velocity profiles have to be known close to the§vessel walls. However, the determination of the exact§vessel geometry is difficult, because the vessels are§very small (10 to 250 micrometres) and almost§transparent. This thesis investigates several§possibilities to stain the endothelium (the cell§layer at the inner boundary of a vessel) of chicken§embryo vasculature, using animals after 2-4 days of§incubation (HH14-17), in order to make it visible§with fluorescence microscopy. The most suitable§staining methods are subsequently used to stain the§endothelium of HH14 chicken embryo hearts and 3D§datasets are acquired by means of confocal§microscopy. Based on these datasets, geometric modelsare reconstructed and, using a numerical flow§simulation package, blood flow-induced wall shear§stress distributions are determined. Subsequently,§the results of the simulations are related to recent§research data about expressions of shear stress§responsive genes of the endothelium.