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，可以洞察细胞的组织和动力学，以及与感染和疾病相关的故障。在过去的25年里，越来越复杂的显微镜，加上新的实验工具，如荧光蛋白标签，改进的激光和相机，以及活细胞成像的发展，导致我们对细胞组织和功能的理解迅速进步。然而，在高分辨率下的成像受到光的性质的限制，约250 nm，这意味着在分子和分子复合物的水平上的信息不能使用传统的光学显微镜获得。因此，高分辨率的研究一直局限于电子显微镜（EM），它只能用于固定和高度加工的样品，并具有有限的能力，以确定特定的分子成分。近年来，已经开发了几种新的光学和计算方法，以打破传统光学显微镜的衍射极限，并有可能以更高的分辨率获得分子/结构信息。2-20倍数改善）。通过将多个荧光标记的探针组合在同一样品内，与活细胞成像一起，这些所谓的超分辨率纳米显微镜技术能够将当前的衍射极限显微镜技术和EM技术连接起来，以产生新的详细的分子和机制见解，从而了解正常细胞功能和疾病的细胞和分子过程。仪器现在变得商业化，使得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、传染病和许多其它生物医学研究领域中的许多问题。开发仍处于早期阶段，仍有许多进步的机会，特别是在活细胞成像、探针开发、软件开发、分析工具等方面。SRI仪器和技术开发人员现在需要与生物医学研究人员密切合作，以建立技术并推动仪器和应用开发。该应用程序将连接细胞生物学单元/分子细胞生物学实验室（CBU/LMCB）和朊病毒单位（MRC PU），以及MRC/UCL分子医学病毒学中心（MRC CMMV），与UCL的生命科学学院，医学和脑科学，伦敦纳米技术中心（LCN），UCL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泰丁顿（NPL）的成员，建立和发展社会责任投资平台。该设施将以商业现成的SRI仪器为中心，这些仪器将提供一个基准，该计划的UCL和NPL成员将开发针对生物医学研究优化的新系统和仪器。该设施还将为学生和博士后研究员提供能力建设机会，在生物学、成像和仪器开发的界面上进行培训/工作。总之，这提供了一个及时的机会窗口，在UCL的优秀成像设施和世界领先的生物医学研究社区的基础上，创建一个前沿的，互动的SRI研究和开发计划，应用于生物医学研究。

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.