CAREER: Imaging the Chemical Nature of Intracellular Phase Separation by Functional Stimulated Raman Scattering Spectro-microscopy

在化学测量和成像（CMI）项目的支持下，生命过程化学（CLP）项目和化学结构、动力学和机制- b （CSDM-B）项目在CHE部门和MCB部门分子生物物理集群的共同资助下，加州理工学院的Lu Wei和她的研究团队正在开发新的光学成像策略，旨在提高我们研究细胞内现象（如与神经退行性疾病相关的现象）的能力。这项研究将产生新的仪器、新的成像探针和新的标记方法，所有这些都是针对活细胞环境中蛋白质和RNA行为的新的化学见解。这个高度跨学科的项目将为研究生和本科生提供研究机会。魏氏小组计划向K-12和本科生提供接触科学的机会。液-液相分离（LLPS）的形成及其在生物功能中的作用已被广泛研究，有许多令人兴奋的新观察和发现。然而，关于生物大分子之间基本相互作用的生物物理和生化原理，特别是蛋白质和RNA及其形成凝聚体的能力，还有很多东西有待学习。为了研究活细胞中关键的细胞内相分离凝聚物，Wei实验室正在开发新的无创、定量的受激拉曼光谱成像策略，并提供高空间和时间分辨率。具体而言，该研究计划旨在(1)开发功能振动探针和标记方法，以量化体外和活细胞研究中鉴定的LLPS系统的组成和感知内部凝聚相互作用；(2)建立新的仪器模块，使活细胞化学制图的灵敏度提高20倍；(3)通过先进的数据分析对活细胞进行深入研究，以拼凑出原生和复杂细胞环境中未知的化学特征和相互作用原理。该奖项由数学和物理科学理事会（MPS）化学部的CMI、CLP和CSDM-B项目以及生物科学理事会（BIO）分子和细胞生物科学部（MCB）的分子生物物理学项目共同资助。该奖项反映了美国国家科学基金会的法定使命，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。

With support from the Chemical Measurement and Imaging (CMI) Program, and co-funding from the Chemistry of Life Processes (CLP) Program and the Chemical Structure, Dynamics, and Mechanisms-B (CSDM-B) Program in the CHE Division and the Molecular Biophysics Cluster in the MCB Division, Lu Wei and her research team at the California Institute of Technology are developing new optical imaging strategies that aim to improve our ability to study intracellular phenomena such as those relevant to neurodegenerative diseases. The research will result in new instrumentation, new imaging probes, and new labeling methods, all targeting new chemical insights into the behavior of proteins and RNA in live-cell settings. The highly interdisciplinary program will provide research opportunities for graduate and undergraduate students. Outreach efforts in the Wei group are planned to provide exposure to science to both K-12 and undergraduate students.The formation of liquid-liquid phase separation (LLPS) and its role in biological function has been extensively studied with many exciting new observations and discoveries. However, there is much yet to be learned especially regarding the biophysical and biochemical principles underlying the fundamental interactions among biomacromolecules, particularly protein and RNA and their ability to form condensates. To investigate key intracellular phase-separated condensates in live cells, the Wei lab is developing new stimulated Raman spectral-imaging strategies that are noninvasive and quantitative, and offer high spatial and temporal resolution. Specifically, the research program aims to (1) develop functional vibrational probes and labeling approaches to quantify the composition and sense the intra-condensate interactions from the identified LLPS systems for both in vitro and live-cell interrogations; (2) build new instrumentation modules to achieve up to 20-fold sensitivity enhancement for chemical mapping in live cells; and (3) perform in-depth live-cell investigations with advanced data analysis to piece together unknown chemical features and interaction principles in native and complex cellular settings.This award is co-funded by the CMI, CLP and CSDM-B Programs in the Division of Chemistry in the Directorate for Mathematical and Physical Sciences (MPS) and by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences (MCB) in the Directorate for Biological Sciences (BIO).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.