Steacie Fellowship RTI - Light sheet microscopy for studies of fractal tubulogenesis

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This RTI application is related to 2014 Steacie Fellowship awarded to Dr. Radisic. The funds are requested to purchase a light sheet microscopy system for the studies of fractal tubulogenesis in tissue engineering of microvasculature. This powerful new technique enables dynamic imaging and tracking of cells in three-dimensional (3D) samples, including living organisms, with minimal photobleaching and phototoxicity. Classical cell biology approaches are limited to the application of single biochemical factors (e.g. growth factors) to cells confined to the surface of a tissue culture plate. A more realistic approach would be to study how cells assemble into complex, branching, 3D structures under stimuli that oscillate or vary in time. Fractals are defined as objects that are self-similar over multiple scales. The branching structure of the circulatory system, consisting of arteries, arterioles, venules and veins, is considered to be fractal. Here, I will develop tissue engineering tools to uncover the complex relationships between heart/pumping rate that varies according to the fractal rules and the fractal geometry of the vasculature. I will then use this knowledge to design shape-memory biomaterials that will simply by their topography and geometry, enable appropriate tissue vascularization. As a model system, I will use vascular progenitors derived from pluripotent stem cells, encapsulated in a collagen/chitosan hydrogel and connected to perfused larger vessels (an artery and a vein) using angiochip, a technology we recently described. Light sheet microscopy will be used to obtain information on fractal dimension of the resulting microvasculature. Repeated on-line imaging of 3D structures is needed for this work, a demand that can be successfully satisfied only by light sheet microscopy. Lack of this equipment will have a serious detrimental effect on the progress of the Steacie Fellowship funded research. Light sheet microscopes are rather scarce in Canada and around the world. At this point, none are available on the main campus of the University of Toronto. Its availability will benefit the 14 HQP in Radisic laboratory, 88 HQP in the collaborating laboratories and colleagues at the University of Toronto and surrounding institutions.