权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Developing a label-free single molecule imaging technology: transitioning from material science to biological science

开发无标记单分子成像技术：从材料科学到生物科学的转变

基本信息

批准号：
1666540
负责人：
金额：
--
依托单位：
University of Kent
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2015
资助国家：
英国
起止时间：
2015 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-1666540
关键词：
Developing label free single molecule

项目摘要

Visualising single proteins in the absence of a label represents a vital step towards studying native protein function. In this application we propose to combine the technologies available at Danish Fundamental Metrology (DFM) with that of the Kad lab (University of Kent) to generate a new label-free imaging tool. This will be used to study DNA repair as a model biological system.The Kad lab presently uses single molecule fluorescence imaging to study nucleotide excision DNA repair (NER) in prokaryotes. This approach has led to considerable new insights into the physical search mechanisms employed by these proteins. To enable further insights we have constructed a high-speed dark-field imaging system that measures the light scattered from individual gold nanoparticle labelled proteins. One key drawback of this system is that the gold nanoparticles (~40 nm) increase the size and thus reduce the diffusion constant of the protein. Therefore we must use theoretical calculations to account for the expected contribution of the attached nanoparticle. Ideally, imaging proteins without a label would provide an accurate measure of how these proteins locate their target sites.We have teamed up with DFM who specialise in measuring the fundamental properties of materials. Such technologies are crucial for advanced optical component development (for example nanostructured gratings). One such system detects light scatter from single unlabelled molecules ordered on a surface (scatterometry). By using scatterometry we will dispense with gold nanoparticles and instead detect the scatter signal from single proteins directly. This technology requires very little hardware development; we use an open optical system that can easily be adapted to accommodate for scatterometry. A major component in this PhD project is the application of Fourier image analysis software, which is the subject of on-going development at DFM.

在没有标记的情况下可视化单个蛋白质是研究天然蛋白质功能的重要一步。在这个应用程序中，我们建议联合收割机的技术，可在丹麦基础计量（DFM）与Kad实验室（肯特大学），以产生一个新的无标签成像工具。Kad实验室目前使用单分子荧光成像来研究原核生物中的核苷酸切除DNA修复（NER）。这种方法导致了相当多的新的见解，这些蛋白质所采用的物理搜索机制。为了实现进一步的见解，我们构建了一个高速暗场成像系统，该系统测量从单个金纳米颗粒标记的蛋白质散射的光。该系统的一个主要缺点是金纳米颗粒（~40 nm）增加了尺寸，从而降低了蛋白质的扩散常数。因此，我们必须使用理论计算来解释附着的纳米颗粒的预期贡献。理想情况下，没有标记的蛋白质成像将提供这些蛋白质如何定位其靶位点的准确测量。我们与专门测量材料基本性质的DFM合作。这些技术对于先进的光学元件开发（例如纳米结构光栅）至关重要。一种这样的系统检测来自表面上有序的单个未标记分子的光散射（散射测量）。通过使用散射测量，我们将免除金纳米颗粒，而是直接检测来自单个蛋白质的散射信号。这项技术需要很少的硬件开发;我们使用一个开放的光学系统，可以很容易地适应散射测量。这个博士项目的一个主要组成部分是傅立叶图像分析软件的应用，这是DFM正在进行的开发的主题。