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组织的宏观水平与细胞和组织的分子甚至原子水平组织之间提供结构理解的无缝链接。由于这些构建模块是纳米级物体，因此将超分辨率显微镜应用于细胞生物学直观上是有意义的。材料科学也在纳米范围内发展，现在很容易生产纳米比例的合成粒子。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。由于细胞反应通常与特定的亚细胞细胞器和室室有关，因此研究颗粒的运输并在亚细胞器水平上高精度地确定其在细胞内的目的地也很重要。几个世纪以来，光学显微镜的一个基本限制是它的分辨率不足以解决支撑生物学的纳米尺度过程。尽管如此，通过许多有机标签的可用性和绿色荧光蛋白的发现，荧光显微镜几十年来一直是生物医学科学中许多基于体外的关键发现的基础。在上个世纪末，用光显微镜的“分辨率极限”被一种具有挑战性的技术——受激发射损耗（STED）显微镜所打破，该技术显示出高达~20纳米的分辨率。近年来，STED显微镜已经从一种专门的、高度专业化的方法，用于对有限的一组合适的样品进行超分辨率成像，发展成为一种广泛的、通用的荧光显微镜模式。STED显微镜现在可以在多色活细胞中进行，甚至可以在视频速率下进行。通过实现非常高的分辨率，STED显微镜为荧光显微镜开辟了一个应用领域，而荧光显微镜以前是电子显微镜的专有领域，其优势在于STED可以在纳米尺度上实时研究几乎任何活细胞细胞器的细胞内生理过程。高分辨率成像是理解基本细胞生物学的关键。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显微镜和一系列相关样品的测试。在显微镜投入使用后，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。科学家们在哈维尔校区的集中将有助于我们在未来十年巩固基础发现的努力。

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.