Unravelling the Working Mechanisms of Fluorescent Molecular Rotors for Bioimaging

揭示生物成像荧光分子转子的工作机制

• 批准号: 1948409
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1948409
• 关键词: Unravelling; Working; Mechanisms; Fluorescent; Molecular

The overall aim of the thesis will be to gain a detailed understanding of the working mechanism of fluorescent molecular rotors, in particular those based on BODIPY. These rotors can be used as probes to evaluate the local viscosity in cell membranes and lipid bilayers. Our goal will be achieved by a combination of multiscale atomistic simulations in combination with advanced fluorescence imaging. The insight gained will allow to optimise the use of fluorescent molecular rotors as rapid and reliable viscosity sensors in fluorescence microscopy, and other biotechnologically relevant viscosity-sensing and imaging assays.The first stage of my project can be summarised by three questions: Is it possible to accurately characterise the potential energy surface of a prototypical BODIPY-based rotor (BODIPY-Phe)? What can we learn about its excited state characteristics? Which simulation methods are the most efficient and effective tools for studying BODIPY-Phe? The aim of this stage of work is, therefore, to refine and compare the simulation techniques which will likely be needed for detailed future studies of the rotor, whilst gaining a qualitative insight into the radiative decay mechanisms that control its photophysical properties.The second stage of the project will build on the first by performing more complex molecular dynamics simulations, both in vacuo and in solvents. These will be done in conjunction with experimental absorption, emission and anisotropy measurements of the BODIPY rotor in solvents of varying viscosities. The aim of these tasks is to gain a more validated understanding of the behaviour of the molecule in real environments. This will all be complemented by computational characterisation of the ground and excited state of two other molecular rotors: DCVJ and ThT. These rotors are thought to have different working mechanisms to BODIPY, so should be good comparative tools.

本论文的总体目标是深入了解荧光分子转子的工作机理，特别是基于BODIPY的荧光分子转子。这些转子可以用作探针，以评估在细胞膜和脂质双层的局部粘度。我们的目标将通过多尺度原子模拟与先进的荧光成像相结合来实现。所获得的洞察力将允许优化使用的荧光分子转子作为快速和可靠的粘度传感器在荧光显微镜，和其他生物技术相关的粘度传感和成像assays.The第一阶段我的项目可以概括为三个问题：是否有可能准确地计算的势能表面的原型BODIPY为基础的转子（BODIPY苯丙氨酸）？关于它的激发态特征，我们可以了解到什么？哪些模拟方法是研究BODIPY-Phe最高效、最有效的工具？因此，这一阶段工作的目的是改进和比较未来详细研究转子可能需要的模拟技术，同时获得控制其物理性质的辐射衰变机制的定性见解。该项目的第二阶段将在第一阶段的基础上进行更复杂的分子动力学模拟，包括在真空和溶剂中。这些将结合实验吸收，发射和各向异性测量的BODIPY转子在不同粘度的溶剂。这些任务的目的是获得对分子在真实的环境中的行为的更有效的理解。这一切都将通过计算表征的基态和激发态的其他两个分子转子：DCVJ和ThT。这些转子被认为具有与BODIPY不同的工作机制，因此应该是很好的比较工具。

项目成果

Fluorescence lifetime imaging for viscosity and diffusion measurements

用于粘度和扩散测量的荧光寿命成像

• DOI: 10.1117/12.2508744
• 发表时间: 2019
• 作者: Economou A

Time-Resolved Fluorescence Anisotropy of a Molecular Rotor Resolves Microscopic Viscosity Parameters in Complex Environments.

• DOI: 10.1002/smll.201907139
• 发表时间: 2020-05
• 期刊: Small
• 影响因子: 13.3
• 作者: I. E. Steinmark;P. Chung;Robert M. Ziolek;Bethan Cornell;Paul E. Smith;J. Levitt;C. Tregidgo