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

Steacie Fellowship RTI - 用于分形管状发生研究的光片显微镜

• 批准号: 461396-2014
• 负责人: Radisic, Milica
• 金额: $ 10.92万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2013
• 资助国家: 加拿大
• 起止时间: 2013-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=542602
• 关键词: Steacie; Fellowship; RTI; Light; sheet

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.

此RTI应用程序与2014年授予Radisic博士的Steacie奖学金有关。本项目拟购置一套光片显微镜系统，用于微血管组织工程中分形小管形成的研究。 这种功能强大的新技术能够动态成像和跟踪三维（3D）样品中的细胞，包括活生物体，具有最小的光漂白和光毒性。经典的细胞生物学方法限于将单一生化因子（例如生长因子）应用于局限于组织培养板表面的细胞。 更现实的方法是研究细胞如何在振荡或随时间变化的刺激下组装成复杂的、分支的3D结构。 分形被定义为在多个尺度上自相似的对象。由动脉、小动脉、小静脉和静脉组成的循环系统的分支结构被认为是分形的。在这里，我将开发组织工程工具，以揭示心脏/泵率之间的复杂关系，根据分形规则和分形几何的血管。然后，我将利用这些知识来设计形状记忆生物材料，这些材料将简单地通过它们的地形和几何形状，使适当的组织血管化。作为一个模型系统，我将使用来自多能干细胞的血管祖细胞，封装在胶原蛋白/壳聚糖水凝胶中，并使用血管芯片（我们最近描述的一种技术）连接到灌注的较大血管（动脉和静脉）。 光片显微镜将用于获得所得微血管的分形维数信息。这项工作需要重复的3D结构的在线成像，只有光片显微镜才能成功地满足这一需求。缺乏这种设备将对斯泰西研究金资助的研究的进展产生严重的不利影响。光片显微镜在加拿大和世界各地都相当稀缺。 目前，多伦多大学的主校区还没有这样的学校。它的可用性将使Radisic实验室的14名HQP，合作实验室的88名HQP以及多伦多大学和周边机构的同事受益。