Super Resolution Imaging for Cell Biology and Neuroscience at UCL

显微镜一直是细胞和分子水平上基于发现的研究的主要工具之一，可以深入了解细胞的组织和动态，以及与感染和疾病相关的缺陷。在过去的25年里，日益复杂的显微镜，加上新的实验工具，如荧光蛋白质标签，改进的激光和照相机，以及活细胞成像的发展，使我们对细胞组织和功能的理解迅速进步。然而，高分辨率的成像受到光的特性限制在~250 nm，这意味着使用传统的光学显微镜无法获得分子和分子络合物水平的信息。因此，高分辨率的研究仅限于电子显微镜(EM)，它只能用于固定的和高度加工的样品，并且识别特定分子组分的能力有限。近年来，一些新的光学和计算方法被开发出来，以打破传统光学显微镜的衍射限制，有可能以显著更高的分辨率(约.提高2-20倍)。通过将同一样本中多个荧光标记的探针与活细胞成像相结合，这些所谓的超分辨率纳米技术能够连接当前的衍射限制显微镜和EM技术，以产生对正常细胞功能和疾病背后的细胞和分子过程的新的详细的分子和机制洞察。超分辨率成像(SRI)仪器现在已经商业化，使细胞生物医学研究人员开始将这些令人兴奋的新技术应用于细胞生物学、神经生物学、传染病和许多其他生物医学研究领域的许多问题。开发仍处于早期阶段，仍有许多取得进展的机会，特别是在活细胞成像、探针开发、软件开发、分析工具等方面。然而，现在需要SRI仪器和技术开发商与生物医学研究人员密切合作，建立技术并推动仪器和应用程序的开发。这一应用程序将把MRC资助的细胞生物学单位/分子细胞生物学实验室(CBU/LMCB)和Prion单位(MRC PU)以及分子医学病毒学(MRC CMMV)的MRC/UCL中心与UCL的生命科学、医学和脑科学学院的成员联系起来，伦敦纳米技术中心(LCN)、伦敦大学学院物理学家和化学家、计算机科学家、基于图像的生物信息学家和国家物理实验室特丁顿(NPL)为SRI建立和开发平台。该设施将集中在商业现成的SRI仪器上，这将提供一个基准，该方案的UCL和NPL成员将根据该基准开发优化用于生物医学研究的新系统和仪器。该设施还将为学生和博士后研究员提供在生物学、成像和仪器开发方面进行培训/工作的能力建设机会。总而言之，这是一个及时的机会之窗，可以在伦敦大学学院卓越的成像设施和世界领先的生物医学研究社区的基础上，创建一个应用于生物医学研究的尖端、互动的SRI研究和开发计划。

Microscopy has been one of the principle tools for discovery-based research at the cell and molecular level, allowing insights to the organisation and dynamics of cells, as well as the faults associated with infection and disease. Over the last 25 years increasingly sophisticated microscopes, together with new experimental tools such as fluorescent protein tags, improved lasers and cameras, and developments in live cell imaging, have led to rapid advances in our understanding of cell organisation and function. Nevertheless, imaging at high resolution is limited by the properties of light to ~250 nm, meaning that information at the level of molecules and molecular complexes cannot be obtained using conventional light microscopes. Thus, high-resolution studies have been restricted to electron microscopy (EM), which can only be used on fixed and highly processed samples, and has limited capacity to identify specific molecular components. In recent years several novel optical and computational approaches have been developed to break the diffraction-limit of conventional light microscopy, with the potential to obtain molecular/structural information at significantly higher resolution (approx. 2-20 fold improvement). By combining multiple fluorescently-labelled probes, within the same sample, together with live cell imaging, these so-called super-resolution nanoscopy techniques are able to bridge the current technologies of diffraction-limited microscopy and EM to generate new detailed molecular and mechanistic insights into the cellular and molecular processes that underlie normal cell function and disease.Super-resolution imaging (SRI) instruments are now becoming commercially available allowing cellular biomedical researchers to begin to apply these exciting new technologies to a multitude of questions in cell biology, neurobiology, infectious disease and many other fields of biomedical research. Development is still at an early stage and there remain many opportunities for progress, particularly in live cell imaging, probe development, software development, analytical tools, etc. However, SRI instruments and technical developers are needed now to work in close collaboration with biomedical researchers to establish the technology and push instrument and application development.This application will link MRC-funded scientists in the Cell Biology Unit/Laboratory for Molecular Cell Biology (CBU/LMCB) and Prion Unit (MRC PU), as well as the MRC/UCL Centre in Molecular Medical Virology (MRC CMMV), with members of UCL's Faculties of Life Sciences, Medicine and Brain Sciences, the London Centre for Nanotechnology (LCN), UCL physicists and chemists, computer scientists, image-based bio-informaticians and the National Physics Laboratory Teddington (NPL), to establish and develop platforms for SRI. The facility will be centred on commercial off-the-shelf SRI instruments that will provide a benchmark against which UCL and NPL members of the programme will develop new systems and instruments optimised for biomedical research. The facility will also provide capacity-building opportunities for students and postdoctoral fellows to train/work at the interface of biology, imaging and instrument development. Together this presents a timely window of opportunity in which to build on UCL's excellent imaging facilities and world leading biomedical research community to create a cutting edge, interactive SRI research and development programme for application to biomedical research.