Super-resolution fluorescence atomic force (SURFACE) microscopy

超分辨率荧光原子力（表面）显微镜

• 批准号: BB/I023518/1
• 负责人: Simon J. Foster
• 金额: $ 15.15万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI023518%2F1
• 关键词: Super; resolution; fluorescence; atomic; force

Bacteria are microscopic organisms of immense importance to humans. Given their diminutive size much of our understanding of how they grow and divide has been determined by a variety of microscopy approaches. Since the earliest microscopes many important discoveries have been made and continued technological developments have permitted new and exciting revelations. Bacterial cell biology and the ability to determine the subcellular localization of components have been largely driven by fluorescence microscopy, whereby individual components are labelled with fluorescent markers. However the limit of detection, coupled with the small dimensions of bacteria has limited the true potential of this approach. Very recently new techniques have allowed fluorescence microscopy to traverse this barrier to attain a level of resolution approaching single molecules. However due to the technical difficulties, lack of commercial availability and expense of some of these developments they have not as yet made a great impact. We have been using a complementary approach called atomic force microscopy, which gives very high resolution of surfaces. We have applied this to the study of bacterial cell walls (the site of action of important antibiotics such as penicillin). This has revealed many important new insights. The aim of the project is to build a new type of microscope coupling super resolution fluorescence and atomic force. We will use our existing microscopy set up as a scaffold to rapidly and efficiently develop the new machine, given the interdisciplinary expertise of the investigator team. The new machine will be tested and further developed within our current bacterial cell wall architecture and dynamics research area. This will provide an ideal framework to develop a new microscope, which can then be utilized and duplicated more widely to allow the rapid uptake of a novel and exciting approach.

细菌是对人类极其重要的微生物。由于它们的体积很小，我们对它们如何生长和分裂的理解主要取决于各种显微镜方法。自从最早的显微镜以来，人们已经有了许多重要的发现，不断的技术发展也带来了新的令人兴奋的发现。细菌细胞生物学和确定组分的亚细胞定位的能力在很大程度上由荧光显微镜驱动，其中单个组分用荧光标记物标记。然而，检测极限加上细菌的小尺寸限制了这种方法的真正潜力。最近的新技术已经允许荧光显微镜穿越这一障碍，以达到接近单分子的分辨率水平。然而，由于技术上的困难，缺乏商业可用性和其中一些发展的费用，他们还没有产生很大的影响。我们一直在使用一种称为原子力显微镜的补充方法，它可以提供非常高的表面分辨率。我们已经将其应用于细菌细胞壁（青霉素等重要抗生素的作用部位）的研究。这揭示了许多重要的新见解。该项目的目标是建立一种新型的超分辨荧光和原子力耦合显微镜。我们将使用我们现有的显微镜设置作为支架，快速有效地开发新机器，考虑到研究团队的跨学科专业知识。新机器将在我们目前的细菌细胞壁结构和动力学研究领域进行测试和进一步开发。这将提供一个理想的框架来开发一种新的显微镜，然后可以更广泛地利用和复制，以允许快速采用一种新的和令人兴奋的方法。

项目成果

Correlative Super-Resolution Optical and Atomic Force Microscopy Reveals Relationships Between Bacterial Cell Wall Architecture and Synthesis in Bacillus subtilis.

• DOI: 10.1021/acsnano.1c04375
• 发表时间: 2021-10-26
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Tank RKG;Lund VA;Kumar S;Turner RD;Lafage L;Pasquina Lemonche L;Bullough PA;Cadby A;Foster SJ;Hobbs JK
• 通讯作者: Hobbs JK