Super-resolution imaging

生物成像技术是利用显微镜对细胞和生物体进行研究的技术，是生命科学研究的关键技术之一。然而，生命运作的方式通常是在固定的样本中研究的，并通过将细胞和生物体简化为二维投影来研究。虽然这种方法已经导致并将在未来导致突破性的发现，但人们意识到，如果可能的话，应该在实时3D环境中研究“生命”。因此，获得活细胞成像，即通过显微镜随时间推移对活细胞或生物体进行研究，对于最先进的生命科学研究至关重要。活细胞成像变得越强大，它可以区分或“解决”的结构越小。可见光的衍射通常将成像系统分辨结构的能力限制在细胞直径的1/50至1/100的尺寸。为了进一步提高分辨率，在过去的几年中已经开发了一些“超分辨率”显微镜方法。然而，这些方法中的许多方法太慢，无法捕捉生命细胞和生物体的快速动态。一种快速的超分辨率方法最近已经商业化，称为结构照明显微镜（SIM）与晶格照明，这是更好地适合于活细胞的成像。我们申请了一个国家的最先进的活细胞成像仪器与晶格SIM，蔡司Elyra 7。它的活细胞超分辨率能力得到了检测固定细胞中更小结构（单分子）的能力的补充。我们将把这项技术整合到沃尔夫森生物成像设备中，这是布里斯托大学生命科学学院的中央显微镜设备。该设施为布里斯托大学的100多名工作人员和数十名外部合作者提供了大量成像技术的访问和技术支持。借助Elyra 7，我们将首次能够提供快速的超分辨率活细胞和单分子显微镜。据我们所知，这将是整个英国西南部的独特能力。Elyra 7将立即由支持该应用程序的布里斯托大学13名重度用户组成的核心小组使用。我们将实施一项详细的战略，以增加在大学，在大西南地区和我们的用户群在行业中的使用。Elyra 7将使大量的研究项目成为可能并得到加强。我们将应用Elyra 7来研究活细胞中的许多研究问题。这些包括亚细胞运输，分子和亚细胞结构（如细胞内的囊泡）的运动，以及需要快速适应我们体内不断变化的生理条件的各种细胞类型（如神经元和血细胞）的行为调节。我们还将使用显微镜来推进我们对合成生物学的理解。我们的一些合成生物学研究重点是设计可用于调节细胞行为的新蛋白质分子，例如作为疫苗的平台。我们的基础研究与转化开发密切相关，通过工业研究合作和一些大学分拆公司。超分辨率显微镜将加强这些合作中的研究，从而产生更大的经济影响。下一代科学家的培训是欧胜生物成像设施的一个重要方面，因为它支持大量的博士。培训方案。由Elyra 7实现的尖端活细胞成像方法的培训将被整合到我们的博士学位中。programmes.许多本科生，从大学和超越，也参加了基于显微镜的研究增强了Elyra 7。Elyra 7的超分辨率能力所带来的惊人图像将加强对中学生和更广泛公众的宣传。

Bioimaging, the investigation of cells and organisms by microscopy, is one of the key technologies for life science research. Often the way life functions has however been studied in fixed samples and by reducing cells and organisms to 2-dimensional projections. Although this approach has led and will also in the future lead to breakthrough discoveries there is a realisation that if possible "life" should be studied in a live 3D environment. Access to live cell imaging, the investigation of live cells or organisms by microscopy over time, thus is critical for state-of-the-art life science research. Live cell imaging becomes the more powerful the smaller the structures are that it can distinguish or 'resolve'. Diffraction of visible light generally limits the ability of imaging system to resolve structures to a size of 1/50th to 1/100th of a cell diameter. For a further gain in resolution a number of 'super-resolution' microscopy approaches have been developed over the last few years. Many of these approaches are however too slow to capture the rapid dynamics of life cells and organisms. A fast super-resolution approach has recently been made commercially available, called structured illumination microscopy (SIM) with lattice illumination, that is much better suited for the imaging of live cells.We apply for a state-of-the-art live cell imaging instrument with lattice SIM, the Zeiss Elyra7. Its live cell super-resolution capability is complemented by the ability to detect even smaller structures, single molecules, in fixed cells. We will integrate the technology into the Wolfson Bioimaging Facility, the centralised microscopy facility of the Faculty of Life Sciences at the University of Bristol. This facility provides access to and technical support for a large number of imaging technologies for 100s of members of staff at the University of Bristol and dozens of outside collaborators. With the Elyra7 we will, for the first time, be able to offer rapid super-resolution live cell and single molecule microscopy. To the best of our knowledge, these would be unique capabilities in the entire Southwest of the UK. The Elyra7 will be used immediately by a core group of 13 heavy users at the University of Bristol that support this application. We will implement a detailed strategy to grow use at the University, in the greater Southwest and in our user base in industry. The Elyra7 will thus enable and enhance a large number of research projects.We will apply the Elyra7 to study many research questions in live cells. These include subcellular trafficking, the movement of molecules and subcellular structures such as vesicles within a cell, and the regulation of the behaviour of various cell types that need to rapidly adapt to changing physiological conditions within our body, such as neurons and blood cells. We will also use the microscope to advance our understanding in Synthetic Biology. Some of our Synthetic Biology research focuses on the design of new protein molecules that can be used to regulate cell behaviour, for example as a platform for vaccines. Our basic research is closely linked to translational exploitation, through industrial research collaborations and a number of University spin out companies. Super-resolution microscopy will strengthen research in these collaborations and therefore generate a larger economic impact.Training of the next generation of scientists is an important aspect within the Wolfson Bioimaging Facility as it supports a large number of Ph.D. training programmes. Training in the cutting-edge live cell imaging approaches enabled by the Elyra7 will be integrated into our Ph.D. programmes. Many undergraduate students, from the University and beyond, also participate in microscopy-based research enhanced by the Elyra7. Striking images enabled by the super-resolution capabilities of the Elyra7 will strengthen outreach to secondary school students and the wider public.