Intracellular Force Generation Measured Through Nuclear Particle Tracking

单个分子马达在细胞内产生非常小的力，但是这些力的协调和整合负责有机体的大规模运动。几十年来，对细胞可以产生的力的研究一直受到广泛关注，但仅限于可视化细胞周围的环境如何响应细胞拉动而变形。然而，通过适当的图像分析工具来消除细胞本身的内在运动，细胞核内的波动也提供了量化细胞内产生的力的度量。这一方法尚处于起步阶段，需要努力充分描述现有技术的特点并与之进行比较。通过确定细胞产生的力，包括相邻细胞是否产生不同的力，可以理解身体的微观单元如何共同作用以形成更大的动态结构。具体而言，这在早期胚胎发育期间的组织形成以及与包括癌症和血管疾病在内的不同疾病状态相关的多细胞功能障碍中是重要的。 该项目将整合本科专业发展与K-3科学推广计划。 PI将开发一个项目，向本科生教授伦理学，重点是生物医学工程师的比较伦理学，对比患者，科学和社会的不同优先事项。通过使用图像分析后处理去除内在的细胞移位和旋转，可以将细胞核内的运动与肌动蛋白-肌球蛋白细胞骨架的力产生相关联。利用这种新开发的核内颗粒跟踪技术，可以测量细胞如何在扩散发生之前的短时间尺度上通过细胞结构传播单层内施加的机械刺激，以及在长时间尺度上相互连接的细胞开始适应细胞结构以响应应力。在多细胞系统内的这种细胞内的力转导测量将允许异质组织系统的牵引力显微镜和其他当代测量具有实验局限性的调查。具体地说，这项研究将包括检查通过一片内皮细胞生长的平滑肌细胞在不同的血管状态和细胞内的C。在胚胎发育过程中。

Individual molecular motors generate very small forces within cells, but the coordination and integration of these forces is responsible for large scale movements of an organism. The study of forces that a cell can generate has been of wide interest for decades but has been limited to visualizing how the environment around the cell deforms in response to the cell pulling. However, with proper image analysis tools for removing intrinsic movements of the cell itself, fluctuations within the cell nucleus also provide a metric for quantifying forces generated inside the cell. This methodology is in its infancy and will require effort to fully characterize and compare to established techniques. By determining the forces that cells generate, including if neighboring cells generate different forces, it is possible to understand how microscopic units of the body work together to form larger dynamic structures. Specifically, this is important in tissue formation during early embryonic development and in multicellular dysfunction associated with different disease states including cancer and vascular disease. The project will integrate undergraduate professional development with K-3 science outreach programs. The PI will develop a program to teach ethics to undergraduate students focusing on Comparative Ethics for Biomedical Engineers contrasting the different priorities of the patient, science and society.By removing intrinsic cellular translocation and rotation using image analysis post-processing, it is possible to correlate movements within the nucleus with force generation of the actin-myosin cytoskeleton. Utilizing this newly developed technique of intranuclear particle tracking it may be possible to measure how cells propagate mechanical stimuli applied within a monolayer through cell structures on short timescales before diffusion can occur and on long timescales as interconnected cells begin to adapt cellular structures in response to stress. This intracellular force transduction measurement within multicellular systems will allow investigation of heterogeneous tissue systems where traction force microscopy and other contemporary measurements have experimental limitations. Specifically, this investigation will include examining force transduction through a sheet of endothelial cells growing on smooth muscle cells in different vascular states in and inside cells of C. elegans during embryo development.