Turnkey video-rate atomic force microscopy for nanometre resolution imaging of functional biomolecules and cellular surfaces

用于功能生物分子和细胞表面纳米分辨率成像的交钥匙视频原子力显微镜

• 批准号: BB/W019345/1
• 负责人: Bart Hoogenboom
• 金额: $ 52.56万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019345%2F1
• 关键词: Turnkey; video; rate; atomic; force

Microscopes are our windows to the cell. Their importance is illustrated by the recent awards of Nobel prizes for super-resolution fluorescence microscopy (2014) and for cryo-electron microscopy (2017). These Nobel prizes also highlight the main types of microscopy that are currently used in biology: fluorescence microscopy, which can provide information on live cells, but at relatively poor resolution (~100s of nm), and electron microscopy, which can provide high-resolution images of biological molecules and cells, but requires samples to be frozen. Using atomic force microscopy, we circumvent some of the limitations of other types of microscopy by effectively tracing the surface of a sample (biological or other) with a very sharp needle, enabling us to image functional biological molecules and living cells as ~1 nm resolution. Since we trace these samples line by line to build an entire image, this is a relatively slow type of microscopy.Here we apply for funding to purchase a so-called high-speed atomic force microscope, which incorporates multiple developments to speed image acquisition up from minutes to less than seconds per frame. We will install the equipment in a well-established user facility and anticipate wide use across different disciplines and institutions. For the short-to-medium term, first experiments we plan are related to protein-DNA interactions that determine DNA packing in the cell, commercial DNA-based nanomaterials developed to facilitate chemical reactions, reshaping of biological membranes, molecular interactions that define how living cells bind to substrates, disruption of bacterial surfaces by next-generation antibiotics, and the molecular structures that define cell shape and mechanics.

显微镜是我们观察细胞的窗口。它们的重要性通过最近的超分辨率荧光显微镜（2014年）和低温电子显微镜（2017年）诺贝尔奖的颁发得到了说明。这些诺贝尔奖还突出了目前在生物学中使用的主要显微镜类型：荧光显微镜，它可以提供活细胞的信息，但分辨率相对较低（约100 nm），电子显微镜，它可以提供生物分子和细胞的高分辨率图像，但需要冷冻样品。使用原子力显微镜，我们通过用非常锋利的针有效地追踪样品（生物或其他）的表面，从而规避了其他类型显微镜的一些限制，使我们能够以约1 nm的分辨率对功能性生物分子和活细胞进行成像。由于我们逐行追踪这些样品以构建整个图像，这是一种相对较慢的显微镜类型。在这里，我们申请资金购买所谓的高速原子力显微镜，该显微镜结合了多项开发，将图像采集速度从每帧几分钟提高到不到秒。我们将在完善的用户设施中安装设备，并预计在不同学科和机构中广泛使用。对于短期到中期，我们计划的第一个实验与蛋白质-DNA相互作用有关，这些相互作用决定了细胞中的DNA包装，开发商业化的基于DNA的纳米材料以促进化学反应，重塑生物膜，定义活细胞如何与基质结合的分子相互作用，下一代抗生素破坏细菌表面，以及定义细胞形状和力学的分子结构。