FastScan atomic force microscope for rapid imaging and property measurement of biological systems under natural conditions.

FastScan原子力显微镜，用于自然条件下生物系统的快速成像和特性测量。

• 批准号: BB/L014904/1
• 负责人: Jamie Hobbs
• 金额: $ 38.97万
• 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: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL014904%2F1
• 关键词: FastScan; atomic; force; microscope; rapid

Atomic force microscopy (AFM) is now starting to deliver its potential for biology, allowing us to follow biological dynamics at the molecular scale in their natural (liquid) environment, to monitor the forces that drive biological processes and locally measure mechanical properties from the sub-protein level to entire tissues. Over recent years in Sheffield we have pioneered the use of this technology to answer questions from protein organisation in photosynthetic membranes, to the architecture of bacterial cell walls and we are expanding into entirely new applications such as the measurement of local mechanical properties in the walls of individual plant cells that are important in enabling their function. However, a new generation of AFMs is now commercially available that provides capabilities that are vital for understanding many of the most pressing problems. Processes at molecular length scales are inherently fast, while AFM, because each pixel of an image is collected consecutively, has traditionally had limited imaging rate. The new fast scanning systems have tackled this problem and are able to follow processes with sub-second image rates. Similarly, single proteins typically operate with very small forces, at the picoNewton level, and the properties of structures such as bacterial cell walls vary subtly over tiny, nanometre, length scales in a way that is important for their function. The new AFMs are able to use smaller cantilevers that are inherently more sensitive to force and less susceptible to noise than those used in conventional microscopes, allowing us to really start to unravel the molecular scale properties of living systems. The purpose of this project is to purchase a fast scanning AFM system and to use it to tackle a diverse range of questions including biological solar energy harvesting, bacterial cell division, dynamic changes as plant epidermal cells respond to environmental changes, and membrane protein function. As well as these initial applications, the Principal Investigator has an excellent track record of building successful collaborations using AFM to tackle biological problems, and the new machine will be vital in further expanding the questions that can be broached. Beyond this, the new microscope will form a vital component of a new centre at the University of Sheffield that will bring together state-of-the-art techniques for AFM, super-resolution optics and cryo-electron microscopy, in combination with internationally leading biology, to really understand living systems from individual molecules to entire cells and tissue. This ambition will occur within an extensive portfolio of BBSRC funded research providing an underpinning facility, vital for our future success.

原子力显微镜(AFM)现在开始发挥其生物学潜力，使我们能够在自然(液体)环境中跟踪分子级别的生物动力学，监控驱动生物过程的力，并局部测量从亚蛋白质水平到整个组织的机械性能。近年来，在谢菲尔德，我们率先使用这项技术回答了从光合膜中的蛋白质组织到细菌细胞壁的结构等问题，我们正在扩展到全新的应用领域，例如测量单个植物细胞壁上的局部机械性能，这对实现它们的功能非常重要。然而，新一代原子力显微镜现在已经可以商业化，它提供了对理解许多最紧迫的问题至关重要的能力。分子长度尺度上的过程本质上是快速的，而原子力显微镜，因为图像的每个像素都是连续采集的，所以传统上成像速度有限。新的快速扫描系统已经解决了这个问题，能够以亚秒的图像速率跟踪过程。同样，单一蛋白质通常在皮牛顿水平上以非常小的力工作，而细菌细胞壁等结构的性质在微小、纳米、长度的尺度上略有不同，这对它们的功能很重要。新型原子力显微镜能够使用更小的悬臂，与传统显微镜中使用的悬臂相比，这些悬臂对力更敏感，对噪音更不敏感，使我们能够真正开始解开生命系统的分子尺度特性。该项目的目的是购买一台快速扫描AFM系统，并将其用于解决一系列问题，包括生物太阳能的获取、细菌细胞的分裂、植物表皮细胞对环境变化的动态变化以及膜蛋白功能。除了这些最初的应用，首席调查员在使用AFM建立成功的合作以解决生物问题方面有着良好的记录，新机器将在进一步扩大可以提出的问题方面发挥关键作用。除此之外，新的显微镜将成为谢菲尔德大学新中心的重要组成部分，该中心将把最先进的原子力显微镜、超分辨率光学和冷冻电子显微镜技术与国际领先的生物学相结合，真正了解从单个分子到整个细胞和组织的生命系统。这一雄心壮志将出现在BBSRC资助的广泛研究组合中，提供对我们未来成功至关重要的基础设施。

项目成果

Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains.

• DOI: 10.1016/j.fm.2016.06.003
• 发表时间: 2016-10
• 期刊: Food microbiology
• 影响因子: 5.3
• 作者: Janganan TK;Mullin N;Tzokov SB;Stringer S;Fagan RP;Hobbs JK;Moir A;Bullough PA
• 通讯作者: Bullough PA

Direct Single-Molecule Observation of Mode and Geometry of RecA-Mediated Homology Search

• DOI: 10.1021/acsnano.7b06208
• 发表时间: 2018-01-01
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Lee, Andrew J.;Endo, Masayuki;Walti, Christoph
• 通讯作者: Walti, Christoph

Direct Imaging of Protein Organization in an Intact Bacterial Organelle Using High-Resolution Atomic Force Microscopy.

使用高分辨率原子力显微镜在完整细菌细胞器中直接对蛋白质组织进行直接成像。

• DOI: 10.1021/acsnano.6b05647
• 发表时间: 2017-01-24
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Kumar S;Cartron ML;Mullin N;Qian P;Leggett GJ;Hunter CN;Hobbs JK
• 通讯作者: Hobbs JK

Cooperative RecA clustering: the key to efficient homology searching.

• DOI: 10.1093/nar/gkx769
• 发表时间: 2017-11-16
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Lee AJ;Sharma R;Hobbs JK;Wälti C
• 通讯作者: Wälti C

Demonstration of the role of cell wall homeostasis in Staphylococcus aureus growth and the action of bactericidal antibiotics.

• DOI: 10.1073/pnas.2106022118
• 发表时间: 2021-11-02
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Salamaga B;Kong L;Pasquina-Lemonche L;Lafage L;von Und Zur Muhlen M;Gibson JF;Grybchuk D;Tooke AK;Panchal V;Culp EJ;Tatham E;O'Kane ME;Catley TE;Renshaw SA;Wright GD;Plevka P;Bullough PA;Han A;Hobbs JK;Foster SJ
• 通讯作者: Foster SJ