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

职业：通过功能受激拉曼散射光谱显微镜对细胞内相分离的化学性质进行成像

• 批准号: 2240092
• 负责人: Lu Wei
• 金额: $ 62.5万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2240092&HistoricalAwards=false
• 关键词: CAREER; Imaging; Chemical; Nature; Intracellular

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.

在化学测量和成像（CMI）计划的支持下，并从CHE部门的化学过程（CLP）计划（CLP）计划（CLP）计划（CLP）计划（CLP）计划，化学结构，动力学和机制B（CSDM-B）计划以及MCB MCB部门的分子生物物理学群集，Lu Wei和她的研究团队在加利福尼亚州的Internion Interrions Intivil Intivies Intivies Intivies Intivies Intivies Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive Insive in例如与神经退行性疾病有关的。这项研究将导致新的仪器，新的成像探针和新的标记方法，所有这些都针对活细胞环境中蛋白质和RNA的行为的新化学见解。高度跨学科的计划将为研究生和本科生提供研究机会。计划在WEI小组中进行外展工作，以向K-12和本科生提供科学的曝光。液态液相分离（LLP）的形成及其在生物功能中的作用已通过许多令人兴奋的新观察和发现进行了广泛的研究。然而，关于生物ac纤维分子之间基本相互作用的基本相互作用的生物物理和生化原理，尤其是蛋白质和RNA及其形成冷凝水的能力，尤其是尤其是有据学习的。为了研究活细胞中的关键细胞内分离的冷凝物，WEI实验室正在开发新的刺激的拉曼光谱图像成像策略，这些策略是无创和定量的，并提供了高空间和时间分辨率。具体而言，该研究计划的目的是（1）开发功能振动探针和标记方法，以量化组成并感知来自已鉴定的LLPS系统的体外和活细胞询问的相关性相互作用； （2）构建新的仪器模块，以实现多达20倍的灵敏度增强活细胞的化学映射； （3）对天然和复杂的细胞环境中的未知化学特征和相互作用原理进行深入的实时调查。该奖项由CMI，CLP和CSDM-B计划在数学和物理科学（MPS）以及分子生物学计划的CMI，CLP和CSDM-B计划中共同资助。对于生物科学（BIO），该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来评估值得支持的。