Photochemical Strategies to Activate Far-Red Fluorescence with Green Light

用绿光激活远红荧光的光化学策略

• 批准号: 1954430
• 负责人: Francisco Raymo
• 金额: $ 58万
• 依托单位: University of Miami
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1954430&HistoricalAwards=false
• 关键词: Photochemical; Strategies; Activate; Far; Red

In this project, funded by the Chemical Structure, Dynamics & Mechanisms B Program of the Chemistry Division, Professor Francisco Raymo at the University of Miami and Professor Hao Zhang at Northwestern University are developing new photochemistry and fluorescence methods to image intracellular components in live cells. This challenging objective first requires the synthesis of a series of structurally related organic dye molecules, which are designed to undergo photoreactions initiated by green light. Photoreaction breaks off a portion of the molecule, thereby generating a new fluorophore, or light emitting molecule, which emits red light. Careful structure design also permits site-specific labeling of intracellular components. The resulting fluorophores enable the imaging of many sub-cellular components in live cells by using high resolution fluorescence microscopy, with spatial resolution at the nanometer level. One goal of these fundamental studies is the identification of optimal structural designs to convert a non-fluorescent reactant into a fluorescent product under mild illumination with green light, a wavelength that is innocuous to live cells. This project is expected to have an immediate impact on photochemistry and fluorescence imaging. Creation of innovative chemical tools to investigate live-cell structures with unprecedented resolution has broader implications in biology and medicine. These research activities provide an outstanding training opportunity for the participating postdoctoral associate, graduate students and undergraduate students. They develop laboratory expertise in chemical synthesis, spectroscopic analysis and fluorescence imaging as well as learn how to culture and manipulate live cells. In the process, they can refine their communication and presentation skills, while also having the opportunity to supervise high-school and undergraduate students in intensive laboratory experiences. This project reinforces an existing student-exchange program with Miami-Dade College, which provides research experiences for students from groups underrepresented in science.Research activities in this project are aimed at the development of a general sensitization mechanism to induce photochemical reactions with green light, thereby providing access to a diverse collection of photocleavable protecting groups with optimal properties for biological applications. New photochemistry is based on modifications to BODIPY (boron-dipyrromethene)-oxazine dyes, which undergo photocleavage of the oxazine component. Dye structures include design elements for spectral tuning, enhancement of aqueous solubility and site-specific attachment to cells. Photoreactions with green light (500 – 530 nm), a wavelength harmless to biological samples, enable bright far-red light fluorescence imaging by generating a new fluorophore. These fundamental studies contribute structural designs to photoactivate fluorescence, under irradiation conditions compatible with live cells, and simultaneously localize multiple spectrally-distinct fluorophores at the single-molecule level with nanoscale precision using single molecule localization microscopy. The unique combination of structural, photochemical and photophysical properties engineered into the proposed compounds can translate into probes with superior performance over the few photoactivatable synthetic dyes developed so far for sub-diffraction imaging of live cells. Thus, this research project, predominantly aimed at the chemical synthesis and spectroscopic analysis of photoswitchable fluorescent constructs, is expected to have an immediate impact on photochemistry and transformative implications in fluorescence imaging.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.

在这个项目中，由化学结构，动力学和机制B计划资助，迈阿密大学的弗朗西斯科·雷莫教授和西北大学的霍张教授正在开发新的光化学和荧光方法，以形象现场细胞中细胞内组件的图像。这个挑战目标首先需要合成一系列与结构相关的有机染料分子，这些分子旨在经历由绿光引发的光反应。光反应会破坏分子的一部分，从而产生新的荧光团或发光分子，该分子发出红光。仔细的结构设计还允许细胞内组件的位点特异性标记。所得的荧光团通过使用高分辨率荧光显微镜将许多亚细胞成分的成像进行成像，并在纳米计水平上进行空间分辨率。这些基本研究的目标之一是确定最佳的结构设计，以将非荧光反应物转换为荧光轻度照明下的荧光产物，这是一种对活细胞无害的波长。预计该项目将对光化学和荧光成像产生直接影响。创建创新的化学工具来研究具有前所未有的分辨率的活细胞结构，对生物学和医学具有更广泛的影响。这些研究活动为参与的博士后同学，研究生和本科生提供了出色的培训机会。他们在化学合成，光谱分析和荧光成像方面发展了实验室专业知识，并学习了如何培养和操纵活细胞。在此过程中，他们可以完善自己的沟通和演示技巧，同时也有机会在密集的实验室经验中监督高中和本科生。项目通过迈阿密戴德学院（Miami-Dade College）加强了现有的学生交往计划，该计划为来自科学领域的人数不足的学生提供了研究经验。该项目的研究活动旨在开发一种具有绿光的普遍敏感性机制，从而诱导光化学反应，从而为具有光学可促进的生物学应用程序提供了绿色光，从而为具有绿色光提供了绿光。新的光化学基于对Bodipy（硼二吡咯甲烯）氧嗪染料的修改，该染料经历了恶敏成分的光丝。染料结构包括用于光谱调整的设计元素，增强水溶性的增强和特定于细胞的位点附件。具有绿光（500 - 530 nm）的光反应，对生物样品无害的波长，通过产生新的荧光团来实现明亮的远红光荧光成像。这些基本研究为在活细胞下的光激活荧光下贡献了结构设计，并在单分子的精度下使用单分子定位显微镜在单分子水平上轻松地将多个光谱荧光团定位。在提出的化合物中设计的结构，光化学和光学特性的独特组合可以转化为探针，其性能优于迄今为止迄今为止为活细胞进行亚分化成像而开发的少数​​光活性合成染料。这项研究项目主要针对可拍照荧光构建体的化学合成和光谱分析，预计将对荧光成像中的光化学和变革性含义产生直接影响。该奖项反映了NSF的法规任务，并认为通过基金会的知识优点和广泛的critia criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criter criteria crietia crietia crietia crietia cristeria crister criter criter criter crite criter corpect。奖励。

项目成果

Photoactivatable Fluorophores for Bioimaging Applications

用于生物成像应用的光活化荧光团

• DOI: 10.1021/acsaom.3c00025
• 发表时间: 2023
• 期刊: ACS Applied Optical Materials
• 作者: Zhang, Yang;Zheng, Yeting;Tomassini, Andrea;Singh, Ambarish Kumar;Raymo, Françisco M.
• 通讯作者: Raymo, Françisco M.

BODIPYs with Photoactivatable Fluorescence

• DOI: 10.1002/chem.202101628
• 发表时间: 2021-06-24
• 期刊: CHEMISTRY-A EUROPEAN JOURNAL
• 影响因子: 4.3
• 作者: Zhang, Yang;Zheng, Yeting;Raymo, Francisco M.
• 通讯作者: Raymo, Francisco M.