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

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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.