High-speed High-throughput AFM For Cell And Developmental Biology

适用于细胞和发育生物学的高速高通量 AFM

• 批准号: BB/R000042/1
• 负责人: Guillaume Charras
• 金额: $ 22.04万
• 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: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR000042%2F1
• 关键词: speed; throughput; AFM; Cell; Developmental

Atomic force microscopy is a form of microscopy in which a tiny needle-like tip is scanned over a surface, thus feeling its contours in a manner similar to that used to read Braille. This technique also functions for work on surfaces immersed in water, which is of great relevance for biology as salty water is the medium that surrounds the minute machinery that makes up living cells, and thus also the human body.The use of Atomic force microscopy is wide-spread in the Physical sciences. However, its use for biology research has been hampered by a number of drawbacks. The aim of this proposal is to create a new-generation atomic force microscopy system that will be uniquely suited to research in cell and developmental biology. The new system will have greatly improved sensitivity and be capable of imaging biological cells and tissues within seconds, compared to minutes for conventional instruments. Importantly, it can go from atomic-resolution imaging of small surfaces to indenting and stretching cells over many microns to determine their mechanical properties. Finally, it can be combined with optical microscopy, which facilitates the detection of fluorescently labelled proteins that be used to report on other phenomena occurring in the cell as it is being probed by the atomic force microscope.These properties make the new-generation atomic force microscopy system highly suitable for addressing a wide range of scientific questions. With our investigators/ collaborators, we have identified five topics for which the application of this system will be most productive:(i) Understand the changes in mechanics that take place during the formation of the spinal chord. Perturbations in the mechanical forces exerted by tissues are at the root of spinal malformations such as spina bifida.(ii) Investigate the mechanical changes that occur during cell division. The answer to this fundamental question has potential implications for future cancer therapies, as tumours are characterised by uncontrolled cell divisions.(iii) Understand how the signals that orchestrate the mechanical changes that occur during cell division.(iv) Determine the physical forces that allow cancer cells to leave a primary tumour and establish secondary tumours in the body.(v) Investigate how pores are formed in the membranes of bacteria and virus infected cells during key functions of the immune response.The instrument we plan to purchase will be located in a user facility at UCL that is widely-used by researchers from all major research-intensive London universities. This will ensure that the expertise gained by the co-investigators will get passed on to the wider AFM user community and that it will benefit biosciences in London at large.

原子力显微镜是显微镜的一种形式，其中微小的针状尖端在表面上扫描，从而以类似于阅读盲文的方式感受其轮廓。这项技术还适用于浸入水中的表面，这与生物学密切相关，因为盐水是构成活细胞乃至人体的微小机械周围的介质。原子力显微镜在物理科学中广泛使用。然而，它在生物学研究中的应用受到了许多缺点的阻碍。该提案的目的是创建新一代原子力显微镜系统，该系统特别适合细胞和发育生物学研究。与传统仪器的几分钟相比，新系统的灵敏度将大大提高，并且能够在几秒钟内对生物细胞和组织进行成像。重要的是，它可以从小表面的原子分辨率成像到将细胞压入和拉伸数微米以确定其机械性能。最后，它可以与光学显微镜相结合，有助于检测荧光标记的蛋白质，这些蛋白质可用于报告原子力显微镜探测细胞中发生的其他现象。这些特性使新一代原子力显微镜系统非常适合解决广泛的科学问题。我们与我们的研究人员/合作者一起确定了应用该系统最有成效的五个主题：(i) 了解脊髓形成过程中发生的力学变化。组织施加的机械力的扰动是脊柱畸形（例如脊柱裂）的根源。(ii) 研究细胞分裂过程中发生的机械变化。这个基本问题的答案对未来的癌症治疗具有潜在的影响，因为肿瘤的特点是不受控制的细胞分裂。（iii）了解如何协调细胞分裂过程中发生的机械变化的信号。（iv）确定允许癌细胞离开原发性肿瘤并在体内建立继发性肿瘤的物理力。（v）研究在细胞的关键功能过程中，细菌和病毒感染细胞的膜上如何形成孔洞。 我们计划购买的仪器将位于伦敦大学学院的用户设施中，该设施被伦敦所有主要研究密集型大学的研究人员广泛使用。这将确保联合研究人员获得的专业知识能够传递给更广泛的 AFM 用户社区，并使整个伦敦的生物科学界受益。

项目成果

Extent of myosin penetration within the actin cortex regulates cell surface mechanics.

• DOI: 10.1038/s41467-021-26611-2
• 发表时间: 2021-11-11
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Truong Quang BA;Peters R;Cassani DAD;Chugh P;Clark AG;Agnew M;Charras G;Paluch EK
• 通讯作者: Paluch EK

Cortical cell stiffness is independent of substrate mechanics.

• DOI: 10.1038/s41563-020-0684-x
• 发表时间: 2020-09
• 期刊: Nature materials
• 影响因子: 41.2
• 作者: Rheinlaender J;Dimitracopoulos A;Wallmeyer B;Kronenberg NM;Chalut KJ;Gather MC;Betz T;Charras G;Franze K
• 通讯作者: Franze K

Cortical cell stiffness is independent of substrate mechanics

皮质细胞硬度与基底力学无关

• DOI: 10.1101/829614
• 发表时间: 2019
• 作者: Rheinlaender J