EAGER: Quantum Imaging for Precise Mapping of Protein Organization during Signaling

细胞中有一种蛋白质叫做受体，它能与信号分子结合并启动生理反应。受体通常是跨膜蛋白，其与细胞外的信号分子结合，随后通过一系列分子开关将信号传递到内部信号通路。目前，还没有可靠的方法来确定信号簇中蛋白质之间的化学计量和精确距离，这对于理解控制信号的基本机制至关重要。本项目中提出的量子增强测量将允许接近精确度的物理极限，以研究蛋白质-蛋白质相互作用并确定细胞信号传导过程中的蛋白质组织。 参与该项目的本科生和研究生将在荧光显微镜、量子光学和精密测量、单光子检测技术、细胞信号和生物学等多个领域获得经验。该项目将展示量子增强测量，以实现完整细胞中蛋白质-蛋白质相互作用的超精密表征。将构建用于两点源高精度成像的图像反转干涉仪系统，并将开发用于量子成像技术所需的质心定位的快速反馈系统。 整合距离测量和传统的单分子超分辨率显微镜（SMSR）信息的分析和估计将被应用于研究完整细胞膜上的EGFR和EGFR/罗恩簇。拟议的工作将开发一种新的技术，将量子增强成像技术与最先进的SMSR相结合，为信号簇中蛋白质的精确成像提供10倍的分辨率增强，分辨率约为1 nm。该项目由生物科学理事会支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Cells have proteins called receptors that bind to signaling molecules and initiate a physiological response. Receptors are generally transmembrane proteins, which bind to signaling molecules outside the cell and subsequently transmit the signal through a sequence of molecular switches to internal signaling pathways. Currently, there are no reliable methods to determine both stoichiometry and precise distances between proteins in a signaling cluster, which is critical for understanding the fundamental mechanisms that control signaling. Quantum-enhanced measurements proposed in this project will allow to approach the physical limits in precision to study protein-protein interactions and determine protein organization during cellular signaling. Undergraduate and graduate students involved in the project will acquire experience in diverse areas including fluorescence microscopy, quantum optics and precision measurements, single-photon detection technologies, cell signaling, and biology.This project will demonstrate quantum-enhanced measurements to realize ultra-precise characterization of protein-protein interactions in intact cells. An image inversion interferometer system for two-point source imaging with high precision will be constructed, and a fast feedback system for center of mass positioning required for the Quantum Imaging technique will be developed. Analyses and estimators that integrate distance measurements and traditional single molecule super resolution microscopy (SMSR) information will be applied to study EGFR and EGFR/Ron clusters on the membrane of intact cells. The proposed work will develop a novel technology that merges quantum-enhanced imaging techniques with state-of-the art SMSR, to provide a 10-fold resolution enhancement for precise imaging of proteins in signaling clusters with approximately 1 nm resolution. This project is supported by the Biological Sciences Directorate.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.