SHeffield IMAging (SHIMA)

从使用第一台显微镜发现细菌，到用神经成像来确定大脑功能，成像一直是生物学和医学认识上的巨大飞跃的核心。所有的进步都完全依赖于新技术的发展。对于肉眼看不到的微小物体的研究尤其如此，因为显微镜是必不可少的。要弄清细胞的功能，无论它们是来自人类、植物、真菌还是细菌，都需要能够看到组成细胞的成分，确定它们的定位，并确定它们如何相互移动。特别是，这是由显微镜的发展推动的，加上使用探针来荧光标记细胞内的分子，从而能够对它们进行定位和跟踪，从而对细胞分裂、生长和分化等过程提供关键的见解。然而，传统的光学显微镜方法的分辨率限制在200纳米左右，这阻碍了对细胞内无数更小的结构和过程的纳米级可视化。直到最近，才发明了新的超分辨率(SR)显微镜方法，使其能够分辨到单分子。因此，我们正处于一个转折点，现有的方法和理解水平将迅速被新的方法所取代。在谢菲尔德，我们很快意识到了这一机会的潜力，我们走在了SR显微镜新方法开发的前沿。这包括建造我们自己的显微镜，将荧光显微镜与其他高分辨率方法结合起来，并将这些进展应用于重要的生物和生物医学问题。因此，作为与应用程序集成的开发计划的一部分，我们处于最佳利用新SR技术的理想位置。谢菲尔德大学(UoS)是一个久负盛名的中心，在生物医学和生物研究的许多方面拥有强大的实力，准备从新的SR技术中获益。我们建议建立一个世界级的SR成像中心，名为谢菲尔德成像(SHIMA)，基于我们目前的优势，拟议的项目和谢菲尔德大学的大笔战略投资。我们将为结构照明显微镜(SIM)和随机光学重建显微镜(STORM)购买商用SR系统，作为允许单分子分辨率和活细胞成像的补充系统。这些设备将安置在我们的光学显微镜设施(LMF)中，以便从一开始就允许多个用户访问。SR是一个新兴领域，因此将建立一个成像和开发中心，以推动技术进步，并将其转移到LMF。为了加强技术开发，我们将实施特定的种子项目，为SIM和STORM生产新的探针，增加技术的多功能性，并提高STORM的速度，使其能够进行3D活细胞成像。与商业伙伴合作，我们将确保我们的发展迅速为社会所用。一个跨学科的团队将管理SHIMA，确保在其直接应用的背景下进行最佳技术开发。两名SHIMA高级实验官员(SEO)将负责跨学科技术开发、新探头的生产和广泛的生物医学应用。SEO将负责培训内部和外部的用户，通过LMF在整个大学(和其他地方)创造一个充满活力的专家研究人员氛围，实现技术开发和应用之间的无缝连接。UOS对Shima的承诺不仅是通过对该项目的大量直接投资，还通过更广泛的成像计划，将谢菲尔德打造为SR成像的关键国际中心。

Imaging has been at the heart of huge leaps in understanding in biology and medicine, from the discovery of bacteria using the first microscopes to neuroimaging to determine brain function. All the advances have been totally dependent on the development of new technologies. This is particularly so for the study of objects too small to be seen by the naked eye, where microscopes are essential. Elucidating how cells function, whether they be from humans, plants, fungi or bacteria requires the ability to be able to see the components that make up the cell, determine their localisation and establish how they move relative to each other. In particular this has been driven by developments in microscopy coupled with the use of probes to fluorescently label molecules inside cells and so be able to localise and track them, giving key insights into processes such as cell division, growth and differentiation. However traditional optical microscopy approaches are limited in resolution to about 200 nm, which prevents the nanoscale visualisation of the myriad smaller structures and processes within the cell. Only very recently have new super-resolution (SR) microscopy approaches been invented to allow resolution down to single molecules. Thus we are at a tipping point, where existing methods and levels of understanding will become rapidly superseded by new approaches. In Sheffield we have been quick to recognise the potential of this opportunity and we are at the forefront of the development of new methods in SR microscopy. This includes building our own microscopes, coupling fluorescence microscopy with other high-resolution approaches and applying these advances to important biological and biomedical questions. We are thus ideally placed to optimally exploit the new SR technology as part of a programme of development integrated with application. The University of Sheffield (UoS) is a well-established centre with great strengths in many aspects of biomedical and biological research that are poised to reap the benefits from the new SR technologies. We propose to establish a world-class centre in SR imaging, entitled Sheffield Imaging (SHIMA), based on our current strengths, the proposed project and a large strategic investment by the University of Sheffield. We will purchase commercial SR systems for both Structured Illumination Microscopy (SIM) and Stochastic Optical Reconstruction Microscopy (STORM) as complementary systems allowing single molecule resolution and live cell imaging. The equipment will be housed in our Light Microscopy Facility (LMF) to allow multiple user access from the outset. SR is an emerging field and so an imaging, development centre will be established to drive forward technological advances, for transfer to the LMF. To enhance technology development we will conduct specific seed projects to produce novel probes for both SIM and STORM, to increase the versatility of the techniques and to increase the speed of STORM to allow live cell imaging in 3D. With commercial partners we will ensure the rapid translation of our developments for the community. An interdisciplinary team will manage SHIMA, ensuring optimal technology development within the context of its direct application. Two SHIMA senior experimental officers (SEOs) will be responsible for interdisciplinary technology development, production of novel probes and establishment of widespread biomedical applications. The SEOs will be responsible for training of users, both internal and external creating a vibrant atmosphere of expert researchers across the University (and beyond) with a seamless interface between technology development and application via the LMF. The UoS is committed to SHIMA not only by a substantial direct investment in the project but also via a wider initiative in imaging to establish Sheffield as a key international centre in SR imaging.