Short FRET: Accurate short distance measurements for synthetic biology and drug discovery.

这个项目的主要目标是：1)开发一个开源的、经济的、强大的、用户友好的单分子测量仪器，任何人都可以建立和使用2)开发新的定量荧光分析，对生物分子结构的微小变化敏感（例如DNA、RNA和蛋白质）。学生将设计，建立和验证一种新的仪器（共聚焦显微镜），能够检测单个荧光分子，一次一个。与本课程中的商业仪器不同，这个新仪器将不以显微镜为基础，学生将研究如何为其他小组提供所有必要的信息来构建仪器。我们预计这种新仪器的价格将是商业版本的一小部分，因此，对许多团体来说，这将是一个有吸引力的选择。第二个目标将是适应当前基于猝灭（可猝灭FRET）的荧光分析，将其发展为完全定量分析。这将涉及分子动力学建模、光物理表征、集合和单分子实验，以确定猝灭的机制，并建立猝灭与生物分子结构之间的关系，这可以被我们和其他人用来观察生物分子的小结构变化，这是目前方法无法实现的。

Democratising Single-molecule FRET and new fluorescence assays with sub 3 nm sensitivityThe key aims of this project are:1) To develop an open-source, economic, robust, and user-friendly instrument for single-molecule measurements, that anyone can build and use2) To develop new quantitative fluorescence assays sensitive to small changes in the structure of biomolecules (e.g. DNA, RNA and proteins) The student will be designing, building and validating a new instrument (a confocal microscope) capable of detecting individual fluorescent molecules, one at a time. Unlike commercial instruments in this class, this new instrument will not be based on a microscope body, and the student will work out how to provide all the necessary information for other groups to build the instrument. We anticipate this new instrument will cost a fraction of the price of commercial versions, and will, therefore, be an attractive alternative for many groups. A second objective will be to adapt a current fluorescence assay based on quenching (quenchable FRET) to develop this into a fully quantitative assay. This will involve molecular dynamics modelling, photo-physical characterisation, and ensemble and single-molecule experiments, to determine the mechanism of quenching and establish a relationship between quenching and biomolecular structure that can be used by us and others to observe small structural changes in biomolecules, which are inaccessible to current methods.