Image correlation studies of distributions and dynamics of macromolecules in cells

新兴的定量生物学和分子医学时代的主要科学挑战之一将是发展对生物大分子如何在活细胞内相互作用以控制生化过程的物理化学理解。目前，只有有限数量的方法可以让科学家直接观察活细胞，并在真实的时间内监测大分子的复杂行为和相互作用。我的研究重点是新的波动显微镜技术应用于活细胞内的大分子的性质的测量。我有兴趣推动使用高时间分辨率阵列检测器相机的基于图像的荧光波动方法的极限。我也有兴趣了解细胞如何在分子水平上调节其信号传导，粘附和迁移。细胞粘附在组织发育和创伤愈合过程中的形成中起着重要作用。它也是某些疾病状态（例如癌细胞转移）的关键过程。我将结合联合收割机荧光显微镜（共聚焦和双光子荧光显微镜）与我们最近开发的时空图像相关光谱（STICS）和空间强度分布分析（SpIDA）的方法和牵引力显微镜（TFM），同时映射力，动力学和负责在活细胞中产生的力的粘附蛋白的聚类。这种原位测量对于解开控制细胞中机械传导的复杂分子机制至关重要。

One of the major scientific challenges in the era of emergent quantitative biology and molecular medicine will be to develop a physico-chemical understanding of how biological macromolecules interact within living cells so as to control biochemical processes. Currently, there are only a limited number of methods that allow scientists to peer directly into a living cell and monitor the complex behavior and interactions of the macromolecules in real time. The focus of my research is novel fluctuation microscopy techniques applied to the measurement of the properties of macromolecules within living cells. I am interested in pushing the limits of image based fluorescence fluctuation methods using high time resolution array detector cameras. I am also interested in understanding how cells regulate their signaling, adhesion and migration at the molecular level. Cell adhesion plays an important role in the formation of tissues during development and wound healing. It is also a pivotal process in certain disease states such as metastasis of cancer cells. I will combine fluorescence microscopy (confocal and two-photon fluorescence microscopy) with our recently developed spatio-temporal image correlation spectroscopy (STICS) and spatial intensity distribution analysis (SpIDA) methods and traction force microscopy (TFM) to simultaneously map forces, dynamics and clustering of adhesion proteins responsible for generating the forces in living cells. Such in situ measurements will be essential for unraveling the complex molecular mechanisms that govern mechanostranduction in cells.