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

生物医学研究的关键工具之一是光学显微镜和更具体的荧光显微镜。荧光显微镜使科学家能够观察细胞、病毒等的标记部分。在分子水平上，它的行为可以被监控。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，从而开发出改进的治疗方法和可能的治愈方法。然而，这些光学显微镜的性能一直受到先前光学分辨率限制的阻碍，该限制是由19世纪德国物理学家恩斯特·阿贝（Ernst Abbe）定义的，因此以他的名字命名，阿贝定律。阿贝定律决定了光学显微镜的最大可实现分辨率是被用来观察它的光的波长（在许多情况下约为550nm）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）。这限制了光学显微镜可以观察生物系统和相互作用的分辨率到100 - 200纳米，随着科学家对生物学的理解的提高，这成为了进一步研究的一个令人沮丧的瓶颈，直到超分辨率或纳米技术的发展和应用。这一新兴领域在2016年因超分辨荧光显微镜的发展而获得诺贝尔化学奖。然而，虽然这些新技术确实允许光学显微镜分辨低至20纳米的图像，但这些技术相当复杂，并不能总是用于科学家可能希望使用的光学显微镜领域的每一种工具。其中一种工具是全内反射荧光显微镜（TIRF）。TIRF目前在细胞生物学中用于观察细胞表面发生的事件，这些事件可以在细胞行为中发挥重要作用，研究癌症的科学家（例如）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TIRF显微镜的空间分辨率提高一倍，使<100nm的特征能够成像。这将为全球科学界提供一种成像系统，可以使用红外射频（SR-TIRF）进行真正的超分辨率活细胞成像，从而进一步推进生物医学研究。

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.