Super Resolution Microscopy in Total Internal Reflection Fluorescence (SR-TIRF)

全内反射荧光超分辨率显微镜 (SR-TIRF)

• 批准号: 104422
• 金额: $ 30.62万
• 依托单位: VISITECH INTERNATIONAL LIMITED
• 依托单位国家: 英国
• 项目类别: Collaborative R&D
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=104422
• 关键词: Super; Resolution; Microscopy; Total; Internal

One of the key tools of bio-medical research are light microscopes and to be more specific fluorescent microscopes. A fluorescent microscope allows the scientist to view tagged parts of a Cell, Virus, etc... at a molecular level so it's behaviour can be monitored.Such fluorescent microscopes are driving modern bio-medical research to aid in our understanding of illness, disease and infection and in turn allow the development of improved treatments and possible cures.However, the performance of these light microscopes has been hampered by the previous limit of optical resolution which was defined by a German Physicist in the 19th Century, Ernst Abbe, and as such carries his name, Abbe's Law.Abbe's Law determined that the maximum achievable resolution of a light microscope is given by the wavelength of the light being used to view it (in many cases around 550nm) divided by twice the numerical aperture of the lens used for imaging (in modern microscopes the Maximum NA = 1.4).This limited the resolution at which light microscope could observe biological systems and interactions to >200nm, as scientists developed an improved understanding of biology this was becoming a frustrating bottle neck for furthering research, until that is the development of super-resolved or Nanoscopy techniques and applications. This new emerging field was highlighted by the 2016 Nobel Price in Chemistry which was awarded for the development of super-resolved fluorescent microscopy.However, whilst these new techniques did allow light microscope to resolve as low as 20nm the techniques were quite complex and could not always be used for every tool within the field of light microscopy which the scientist may have wished to use.One such tool is Total Internal Reflection Fluorescence (TIRF) Microscopy. TIRF is currently used in Cell biology to view events which happen at the surface of cells, such events can play a major roll in the behaviour of the cell and scientists who research cancer (for example) are trying to understand the movement and spread of cancer cells by looking at events at the cellular surface using TIRF microscopes.The instrument we propose to develop within this project will double the spatial resolution of a regular TIRF microscope enabling features <100nm to be imaged. This will offer the global science community an imaging system which can do true super-resolved live cell imaging with TIRF (SR-TIRF) allowing further advancements within bio-medical research.

生物医学研究的关键工具之一是光学显微镜，更具体地说是荧光显微镜。荧光显微镜允许科学家查看细胞，病毒等的标记部分。这种荧光显微镜正在推动现代生物医学研究，以帮助我们了解疾病，疾病和感染，并反过来允许改进治疗方法和可能的治愈方法的发展。然而，这些光学显微镜的性能受到了以前的光学分辨率限制的阻碍，这是由一位德国物理学家在19世纪世纪定义的，恩斯特·阿贝，以他的名字命名，阿贝定律：阿贝定律确定光学显微镜的最大分辨率是由观察它的光的波长决定的（在许多情况下大约550 nm）除以用于成像的透镜的数值孔径的两倍（在现代显微镜中，最大NA = 1.4）。这将光学显微镜观察生物系统和相互作用的分辨率限制在> 200 nm，随着科学家对生物学的理解不断提高，这成为进一步研究的一个令人沮丧的瓶颈，直到超分辨或纳米显微镜技术和应用的发展。2016年诺贝尔化学奖的获奖原因是超分辨荧光显微镜的发展，这一新兴领域受到了关注。虽然这些新技术确实允许光学显微镜分辨低至20 nm的光，但是这些技术相当复杂，并且不能总是用于科学家可能希望使用的光学显微镜领域内的每一种工具。反射荧光（TIRF）显微镜。TIRF目前用于细胞生物学，以观察细胞表面发生的事件，这些事件对细胞的行为和研究癌症的科学家起着重要作用（例如）我们正试图通过使用TIRF显微镜观察细胞表面的事件来了解癌细胞的运动和扩散。我们建议在这个项目中开发的仪器将使常规仪器的空间分辨率增加一倍。TIRF显微镜能够对<100 nm的特征进行成像。这将为全球科学界提供一种成像系统，可以使用TIRF（SR-TIRF）进行真正的超分辨率活细胞成像，从而进一步推动生物医学研究的发展。