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。最近获得的超分辨率荧光显微镜奖（2014 年）和冷冻电子显微镜奖（2017 年）诺贝尔奖说明了它们的重要性。这些诺贝尔奖还强调了目前生物学中使用的主要显微镜类型：荧光显微镜，可以提供活细胞的信息，但分辨率相对较低（约100纳米），以及电子显微镜，可以提供高分辨率生物分子和细胞的图像，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）的表面来规避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本以构建整个图像，因此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，该显微镜结合了多种开发来加速图像采集每帧从几分钟到不到几秒。我们将把这些设备安装在完善的用户设施中，并预计在不同学科和机构中广泛使用。中短期内，我们计划的第一个实验涉及蛋白质-DNA 相互作用（决定细胞中的 DNA 堆积）、开发用于促进化学反应的商业 DNA 纳米材料、生物膜的重塑、定义生命如何进行的分子相互作用。细胞与基质结合、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。