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

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。这种技术也可以用于浸没在水中的表面上的工作，这与生物学非常相关，因为盐水是组成活细胞的微小机器周围的介质，因此也是人体。原子力显微镜在物理科学中的应用非常广泛。然而，它在生物学研究中的应用受到许多缺点的阻碍。该提案的目的是创建一个新一代的原子力显微镜系统，该系统将特别适合于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压痕和拉伸许多微米的细胞，以确定它们的机械性能。最后，它还可以与光学显微镜结合使用，便于检测荧光标记的蛋白质，从而在原子力显微镜探测细胞时报告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。与我们的研究人员/合作者，我们已经确定了五个主题，该系统的应用将是最富有成效的：（i）了解在脊髓形成过程中发生的力学变化。由组织施加的机械力的扰动是脊柱畸形如脊柱裂的根源。(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，因为肿瘤的特征是不受控制的细胞分裂。(iii)了解细胞分裂过程中发生的机械变化的信号是如何协调的。(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)研究在免疫反应的关键功能过程中，细菌和病毒感染细胞的细胞膜是如何形成孔的。我们计划购买的仪器将位于伦敦大学学院的一个用户设施中，该设施被所有主要研究密集型伦敦大学的研究人员广泛使用。这将确保共同研究者获得的专业知识将传递给更广泛的AFM用户社区，并使伦敦的生物科学受益。

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.