Decipher membrane patterns in situ with super-resolution and dynamic microscopy

描述（由申请人提供） 翻译后摘要：目前的超分辨率方法无法生成多色原位图像和动态信息限制了它们的影响。目前的挑战是推进这种纯粹描述性的技术，以研究高度动态和罕见的分子过程的机制。我们将通过不同的荧光团和新的采集策略的组合来实现活标本中两种不同颜色的可视化。分子分布和运动之间的相互作用的信息将来自荧光互相关光谱的分子动力学的同时分析。利用这种新技术，我们将研究质膜的结构及其对膜相关信号的影响。这项研究将提供必要的视觉和机械的见解，发展一个统一的质膜理论。具体来说，我们将探讨所有膜蛋白到域的隔离和其独特的限制的基础。此外，本研究还将探讨不同信号通路（TCR、EGF和细胞因子信号通路）的时空机制。我们希望发现新的控制原则，在信号是植根于质膜的架构。因此，我们的研究有可能彻底改变我们对调节膜相关信号的分子基础的理解。鉴于大量疾病与膜信号传导缺陷相关，这是至关重要的。我们的研究将验证通过质膜组织的变化来调节膜相关信号的潜力。 公共卫生相关性：研究质膜结构和植根于其中的信号传导机制为深入了解由膜信号传导缺陷引起的大量疾病奠定了基础。拟议的研究旨在建立一种技术，可以基于超分辨率和互相关光学显微镜确定膜相关信号的空间和动态基础。它将提供新的途径，通过改变其环境来调节细胞信号，这是一种更“温和”的调节，而不会引起当前方法的显著副作用。

DESCRIPTION (Provided by the applicant) Abstract: The inability of current super-resolution methods to generate multi color in situ images and dynamic information limits their impact. The challenge at hand is to advance this purely descriptive technology to study the mechanisms of highly dynamic and rare molecular processes. We will approach the task of visualizing two different colors in live specimens through the combination of different fluorophores and new acquisition strategies. Information on the interplay between molecule distributions and movement will be derived from simultaneous analyses of molecule dynamics by fluorescence cross correlation spectroscopy. With this novel technology, we will study the architecture of the plasma membrane and its effects on membrane associated signaling. This research will provide visual and mechanistic insights necessary to develop a unified plasma membrane theory. Specifically, we will explore the segregation of all membrane proteins into domains and the basis of their distinct confinement. In addition, the proposed research wil disect the spatio-temporal mechanisms of different signaling pathways (TCR, EGF and cytokine signaling). We expect to discover novel control principles in signaling that are rooted in the architecture of the plasma membrane. Thus, our studies have the potential to revolutionize our understanding of the molecular underpinnings regulating membrane- associated signaling. This is crucially important in view of the large number of diseases associated with membrane signaling defects. Our studies will verify the potential for the modulation of membrane associated signaling through changes in the plasma membrane organization. Public Health Relevance: Studying the plasma membrane structure and signaling mechanisms rooted in it lays the foundation for insights into a large number of diseases that are caused by defects in membrane signaling. The proposed research aims to establish technology that can determine the spatial and dynamic underpinnings of membraneassociated signaling based on super-resolution and cross-correlation optical microscopy. It will provide new avenues for modulating cellular signaling through changes in its environment, a more 'gentle' adjustment without causing the dramatic side effects of current approaches.