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.

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.这项研究的目的是探索这些创新的荧光染料的独特特性，以研究以前所未有的分辨率研究细胞过程，并最终贡献了宝贵的化学工具，以阐明负责正常细胞功能和疾病的基本因素。该项目位于有机和物理化学的界面，对生物学和医学的长期影响。拟议的活动是本科生，研究生和博士后同事的最佳培训场，并通过在这两个机构提供既定的外展计划，参与高中生参加密集实验室经验的培训。该项目还将涉及在当地社区学院的机构和协作宣传工作的参与。该项目的目的是生成一个可渗透性和可腐烂的可渗透性和可生物偶联的合成染料的图书馆，并具有光化的光化染料，其设计基于对绿bodipipyrometheene的绿色效果的设计，以实现其设计。周期。他们的实现需要合成，结构表征，集合/单分子光谱分析以及四个光活性Bodipys的成像，在两个平行项目组件中，其多环子的彩色支架的环形融合模式和亚属性的性质不同。一个人旨在解决光活化荧光的光谱位置和/或带状形状，以使单分子发射光谱分化充分以统计置信度以结构性区分。另一个旨在在发色平台上引入亲水基团并靶向配体，以选择性地标记和跟踪细胞内蛋白质。总体科学的目标是最终将结构，光化学和光学物理特性与创新分子结构的独特组合进行设计，以使实时细胞中蛋白质的前所未有的高通量多重单分子跟踪实时奖励的奖励均通过评估良好的依据，这表明了NSF的概述。