A sharper light from gSTED microscopy on biological structure and molecular interactions

21世纪细胞生物学家的中心目标之一是在组织的宏观组织水平与细胞和组织的构建块的分子甚至原子水平的组织之间提供结构理解的无缝联系。由于这些构件是纳米级的物体，将超分辨率显微镜应用于细胞生物学是有意义的。材料科学也已经发展到纳米级，现在很容易就能生产出纳米级的人造颗粒。研究这些颗粒及其与同样小的细胞成分的相互作用在许多生物医学应用中也变得重要，从靶向治疗到骨修复。由于细胞反应往往与特定的亚细胞细胞器和隔间有关，因此研究颗粒的运输并准确地定位它们在细胞内的目的地也是很重要的，即使在亚细胞器水平上也是如此。几个世纪以来，光学显微镜的一个基本局限性是，分辨率不足以分辨支撑生物学的纳米级过程。尽管如此，由于许多有机标记的出现和绿色荧光蛋白的发现，荧光显微镜几十年来一直是许多基于体外的生物医学科学关键发现的基础。上个千年末，一种具有挑战性的技术--受激发射耗尽(STED)显微镜打破了光学显微镜的“分辨率极限”，它的分辨率可达~20 nm。近年来，STED显微镜已经从一种对有限类型的合适样品进行超分辨率成像的专有和高度专业化的方法发展到一种广泛的、通用的荧光显微镜模式。STED显微镜现在可以在多种颜色的活细胞中进行，甚至可以视频率进行。通过获得非常高的分辨率，STED显微镜开辟了荧光显微镜的应用领域，而荧光显微镜以前是电子显微镜的专有领域，其优势是STED允许实时研究活细胞几乎任何细胞器中纳米级的细胞内生理过程。高分辨率成像是了解基础细胞生物学的关键。我们已经形成了一个跨学科的合作伙伴关系，寻求在多学科环境中利用STED显微镜和在一系列相关样本中进行测试。在调试完显微镜后，我们的经验将帮助其他科学家和合作者应用这种方法来回答他们的科学问题。科学家们集中在哈威尔校区，将有助于我们在未来十年巩固基本发现的努力。

One of the central goals of 21st century cell biologists is to provide a seamless link of structural understanding between the macroscopic level of tissue organization to the molecular and even atomic level organization of the building blocks of cells and tissues. As these building blocks are nanoscale objects, applying superresolution microscopy to cell biology intuitively makes sense. The material sciences have also advanced in to the nanometre range as well, readily producing nowadays synthetic particles of nanometre proportions. Studying these particles and their interactions with equally small cellular components has also become important in many biomedical applications, from targeted therapeutics to bone repair. As cellular reactions are often associated with specific subcellular organelles and compartments it is also important to study the transport of particles and pinpoint their destination within the cell with high accuracy even at the sub-organelle level.For centuries a fundamental limitation of light microscopy was that is resolution was insufficient to resolve the nanoscale processes underpinning biology. Despite this, through the availability of many organic labels and the discovery of green fluorescent protein, fluorescence microscopy has been fundamental for decades to many of the in vitro-based key discoveries in the biomedical sciences. The 'resolution limit' of light microscopy was broken at the end of the last millennium using a challenging technique, stimulated emission depletion (STED) microscopy, which showed up to ~20 nm resolution. In recent years, STED microscopy has matured from an exclusive and highly specialised method for superresolution imaging of a limited set of suitable sample types, to a widespread, general purpose mode of fluorescence microscopy. STED microscopy can now been performed in multi-colour, live cells and even at video rate. By achieving very high resolution, STED microscopy has opened up a field of application for fluorescence microscopy that had been previously been an exclusive domain of electron microscopy, the advantage being that STED allows investigating intracellular physiological processes in the nanoscale in almost any organelle of a living cell, and in real time. High resolution imaging is critical to understand basic cell biology. We have formed an interdisciplinary partnership that seeks to exploit STED microscopy and testing in a range of relevant samples within a multidisciplinary environment. After commissioning the microscope, our experience will help other scientists and collaborators to apply this method to answer their scientific questions. The concentration of scientists at the Harwell Campus will help in our efforts to underpin fundamental discoveries in the next decade.