A super-resolution microscopy platform for imaging cells at multiple spatial scales

用于在多个空间尺度上成像细胞的超分辨率显微镜平台

• 批准号: BB/X019845/1
• 负责人: Juan Burrone
• 金额: $ 64.39万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX019845%2F1
• 关键词: super; resolution; microscopy; platform; imaging

Fluorescence imaging has allowed researchers to explore the localisation, dynamics and plasticity of structures within cells. Although revolutionary at the time, the resolution of classical fluorescence imaging microscopes is constrained by the diffraction limit of light. This means that the precise location and organisation of molecules in a cell cannot be accurately visualised. Instead, only a blurred description of their whereabouts was possible, without any possibility of uncovering how single molecules were arranged in space. The advent super-resolution microscopy has changed all this - it is now possible to resolve the location of individual molecules using fluorescence imaging and begin to explore a whole new world of molecular arrangements that takes place at the nanoscale. In fact, these new approaches have been instrumental in uncovering novel sub-cellular structures within cells, which will help better understand their function. To delve into this new world of the very small, it is important to have the right equipment that allows single-molecule imaging. In particular, it needs to be able to do so for molecules tagged with different fluorescent compounds to uncover multi-molecular structures, with high precision in 3D and over large fields of view to encompass as much information as possible. Finally, it is important to also have access to imaging tools that give low resolution maps of the molecules of interest. In this way, we aim to provide large-scale descriptions of the distribution of molecules in cells and neurons and subsequently home into some of these domains to characterise them at very high spatial resolution. Here, we propose to use this approach to answer basic cell biology questions in different systems, including in intact tissue slices, to establish the properties of synapses formed by neurons in the brain and understand the local structure of the cytoskeleton in cells grown in vitro.

荧光成像使研究人员能够探索细胞内结构的定位，动力学和可塑性。虽然在当时是革命性的，但经典荧光成像显微镜的分辨率受到光的衍射极限的限制。这意味着细胞中分子的精确位置和组织无法准确可视化。相反，只能模糊地描述它们的下落，而不可能揭示单个分子在空间中的排列方式。超分辨率显微镜的出现改变了这一切-现在可以使用荧光成像来解析单个分子的位置，并开始探索在纳米级发生的分子排列的全新世界。事实上，这些新方法有助于揭示细胞内新的亚细胞结构，这将有助于更好地了解它们的功能。为了深入研究这个非常小的新世界，拥有允许单分子成像的正确设备是很重要的。特别是，它需要能够这样做，以不同的荧光化合物标记的分子，以揭示多分子结构，在3D和大视场中具有高精度，以涵盖尽可能多的信息。最后，重要的是还可以使用成像工具，这些工具可以提供感兴趣分子的低分辨率图。通过这种方式，我们的目标是提供细胞和神经元中分子分布的大规模描述，并随后进入其中一些领域，以非常高的空间分辨率对它们进行定位。在这里，我们建议使用这种方法来回答不同系统中的基本细胞生物学问题，包括在完整的组织切片中，以建立由大脑中的神经元形成的突触的特性，并了解体外生长的细胞中细胞骨架的局部结构。