Understanding Rotational Dynamics of Cargo in Intracellular Transport with Plasmonic Nanoparticle Probes

方1004612A细胞可以被设想为一个包含纳米机器网络的工厂。许多这样的纳米机器以循环的方式将化学能转化为机械功。这些纳米机器中的任何一个发生故障都可能导致严重的疾病。卫生和医药领域迫切需要新的创新技术来研究活细胞中纳米机器的组成、动力学和工作机制。平移运动可以很容易地通过各种传统的单粒子跟踪(SPT)方法来揭示。然而，由于具有挑战性的技术限制，活细胞中的旋转运动更难解决，而且在很大程度上是未知的。拟议的研究结果将为开发治疗细胞纳米机械故障引起的疾病奠定基础，这些疾病包括某些类型的失明和肾脏疾病，以及神经退行性疾病和寄生虫病。这项研究将被整合到课程中，并将包括本科光学成像实验室实验。PI建议开发一种基于单粒子定向和旋转跟踪(SPORT)的新成像系统，这是PI发明的一项技术。该系统将提供有关粒子的5维信息(xyz位置加上两个角方向)，以更好地了解细胞内的货物运输。PI将开发光学显微镜方法来跟踪附着在细胞内货物上的单个金纳米棒的旋转，如囊泡和马达蛋白，当与x、y和z平移相结合时，将提供更全面的了解它们在活细胞中的动力学。

Fang1004612A cell can be conceived as a factory containing a network of nanomachines. Many of these nanomachines transduce chemical energy into mechanical work in a cyclic manner. The malfunction of any of these nanomachines can lead to severe disease. There is an urgent need from the health and medicine sector for new innovative techniques to investigate the composition, dynamics, and working mechanism of the nanomachines in live cells. Translational motion can be readily revealed by a variety of conventional single particle tracking (SPT) methods. However, rotational motion in live cells is much more difficult to resolve and largely unknown due to challenging technical limitations. The outcomes of the proposed research will lay the groundwork for development of treatments for conditions caused by the malfunction of the cellular nanomachinery, ranging from certain kinds of blindness and kidney disease to neurodegenerative disorders and parasitic diseases. The research will be integrated into the curriculum and will include undergraduate optical imaging laboratory experiments. The PI proposes to develop a new imaging system based on single particle orientation and rotational tracking (SPORT), a technology invented by the PI. This system will provide 5 dimensional information on particles (xyz location plus two angular orientation) to better understand intracellular cargo transport. The PI will develop optical microscopy methods to track the rotation of single gold nanorods attached to intracellular cargo, such as vesicles and motor proteins, which when combined with x,y, and z translations will offer more complete insight into their dynamics in live cells.