Collaborative Research: Spectral Discrimination of Single Molecules with Photoactivatable Fluorescence

合作研究：利用光激活荧光对单分子进行光谱辨别

• 批准号: 2246548
• 负责人: Yang Zhang
• 金额: $ 20.91万
• 依托单位: North Carolina State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2246548&HistoricalAwards=false
• 关键词: Collaborative; Research; Spectral; Discrimination; Single

With support of the Chemical Structure, Dynamics & Mechanisms-B (CSDM-B) and Chemical Measurement and Imaging (CMI) Programs of the Chemistry Division, Françisco M. Raymo of the Department of Chemistry at the University of Miami and Yang Zhang of the Department of Textile Engineering at North Carolina State University are developing a new class of synthetic dyes with photoactivatable fluorescence for bioimaging applications. The goal of this research is to exploit the unique properties of these innovative fluorescent dyes to investigate cellular processes with unprecedented resolution and, ultimately, contribute invaluable chemical tools to elucidate the fundamental factors responsible for normal cellular functions and disease inception. The project lies at the interface of organic and physical chemistry with long-term implications in biology and medicine. The proposed activities are an optimal training ground for undergraduate, graduate students and postdoctoral associates as well as involve the participation of high-school students in intensive laboratory experiences through established outreach programs available at both institutions. The project will also involve participation of members of underrepresented groups at both institutions and collaborative outreach efforts at a local community college.The goal of this project is to generate a library of cell-permeable and bioconjugatable synthetic dyes with photoactivatable fluorescence and resolved single-molecule emission spectra. Their design is based on a photochemical mechanism to activate borondipyrromethene (BODIPY) fluorescence with green light as developed by the team in prior funding cycles. Their realization demands the synthesis, structural characterization, ensemble/single-molecule spectroscopic analysis and imaging of four series of photoactivatable BODIPYs, differing in the ring-fusion pattern of their polycyclic chromophoric scaffold and/or nature of their substituents, in two parallel project components. One is aimed at resolving the spectral position and/or band shape of the photoactivated fluorescence to differentiate the single-molecule emission spectra sufficiently for structural discrimination with statistical confidence. The other is aimed at introducing hydrophilic groups and targeting ligands on the chromophoric platform to label selectively and track simultaneously intracellular proteins. The overarching scientific goal is to ultimately engineer of a unique combination of structural, photochemical and photophysical properties into innovative molecular constructs to enable the unprecedented high-throughput multiplexed single-molecule tracking of proteins in live cells.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.

在化学系化学结构、动力学与机理-B（CSDM-B）和化学测量与成像（CMI）项目的支持下，迈阿密大学化学系的Françisco M. Raymo和迈阿密大学化学系的张扬北卡罗来纳州立大学纺织工程系正在开发一种具有光活化荧光的新型合成染料，用于生物成像应用。这项研究的目标是利用这些创新荧光染料的独特特性进行研究。该项目以前所未有的分辨率研究细胞过程，并最终为阐明正常细胞功能和疾病发生的基本因素提供了宝贵的化学工具，对生物学和医学活动具有长期影响。为本科生、研究生和博士后提供了一个最佳的培训场地，并通过两个机构提供的既定外展计划让高中生参与强化实验室体验。该项目还将让两个机构中代表性不足的群体成员参与其中。以及在当地的协作外展工作该项目的目标是生成一个具有可光激活荧光和解析单分子发射光谱的细胞渗透性和可生物共轭合成染料库。其设计基于用绿光激活二吡咯亚甲基 (BODIPY) 荧光的光化学机制。正如该团队在之前的资助周期中开发的那样，他们的实现需要四个系列的光活化 BODIPY 的合成、结构表征、整体/单分子光谱分析和成像，在两个平行项目组件中，其多环发色支架的环融合模式和/或取代基的性质不同，其中一个旨在解析光活化荧光的光谱位置和/或带形状以区分单分子发射。另一个目标是在发色平台上引入亲水基团和靶向配体，以选择性地标记并同时跟踪细胞内蛋白质。将结构、光化学和光物理特性独特地组合到创新的分子结构中，以实现对活细胞中蛋白质的前所未有的高通量多重单分子追踪。该奖项反映了 NSF 的法定使命，并通过使用基金会的评估进行评估，被认为值得支持。智力价值和更广泛的影响审查标准。