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

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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.