Stimulated Emission Depletion Microscopy (STED) for imaging at high resolution in the Biosciences

为了了解蛋白质在细胞和组织中是如何组织的，以及它们如何相互作用，我们需要能够以高水平的细节看到细胞内部。不幸的是，由于光学显微镜的工作方式，我们通常可以看到的细节是有限的。在这个提案中，我们希望建立一种新型的显微镜，它克服了传统光学显微镜的一些缺点，并将使我们能够看到更多的细节，将模糊的图像变成更清晰的图像。通常情况下，我们只能分辨一个细胞中的两个物体，如果它们相距超过250纳米。这种新的显微镜将使我们能够分辨细胞中相距超过30到40纳米的两个物体，使我们可以看到的细节增加约10倍。这将使我们能够找到一些重要问题的答案，比如细胞骨架在不同类型的细胞中是如何组织的，它是如何在初级纤毛中采用专门的组织的。（细胞表面的小触角状结构），病毒如何在细胞内复制，膜蛋白如何在细胞表面组织，蛋白质如何在心肌细胞中组织，等等。这种新的显微镜将给我们的成像能力带来一个飞跃，我们将能够看到的新的细节水平将为广泛的生物和健康相关研究提供基础。

To find out how proteins are organised within cells and tissues, and how they interact with each other, we need to be able to see inside the cell with a high level of detail. Unfortunately, because of the way light microscopes work, the amount of detail we can normally see is limited. In this proposal, we want to set up a new type of microscope, that overcomes some of the shortcomings of traditional light microscopes, and will allow us to see much more detail, turning a blurry image into a much sharper one. Normally, we can only resolve two objects in a cell if they are more than 250 nanometers apart. This new microscope will allow us to resolve two objects in a cell that are more than ~30 to 40 nm apart, increasing the amount of detail we can see by about 10 fold. This will enable us to find the answers to important questions about how the cytoskeleton is organised in different types of cell, how it adopts a specialised organisation in primary cilia (small antennae like structures on the cell surface), how viruses replicate inside cells, how membrane proteins are organised on the surfaces of cells, how proteins are organised in heart muscle cells, and so on. This new microscope will bring a step change to our imaging capabilities, and the new level of detail that we'll be able to see will underpin a wide range of biological and health related research.