Liverpool BioAFM: an integrated optical and atomic force microscope for research across the life sciences

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Individual cells in a plant or an animal are exposed to changes in their environment (mechanical forces, biochemical signals, temperature, light variations...). Cells have to interpret this information to adapt and respond appropriately. To understand the molecular mechanisms leading to a particular response (e.g. cell death, differentiation, adhesion, changes in topology etc), biologists need means of manipulating the environment in a carefully controlled manner and measuring the effects, for instance on the levels and localisation of proteins inside cells or on the structural properties of the cell surface. Each individual cell might respond differently from its neighbour and at a different time so it is crucial to follow the events in real time and in each individual cell. This can be achieved using BioAFM imaging, an emerging technology, which combines Atomic Force Microscopy (AFM) with advanced optical imaging (e.g. confocal, TIRF). AFM is one of the foremost tools for imaging, measuring, and manipulating matter at the nanoscale. Information is gathered by "feeling" the surface with a mechanical probe. However, this technique does not provide information about the events inside cells, which can only be imaged using advanced optical imaging using fluorescently labelled molecules. Although BioAFM techniques have existed for over a decade in specialist centres, technological limitations restricted the ability to readily combine measurements from these complementary approaches. Only now that the technology has advanced and been refined through the development of user-friendly routines to capture and overlay information has it reached the point where it can be made available more widely to the life science community.We propose to purchase one of the first BioAFM microscopes in the UK and to install it in the Liverpool Centre for Cell Imaging (CCI). The CCI is an open access facility, so the microscope will be accessible to groups from several universities and companies. To illustrate the breadth of the science that will be served by this equipment, we briefly present below three of our exemplar projects:1. Novel surfaces for anti-microbial resistanceRepeated warnings from the WHO and the UK Government's Chief Medical Officer emphasise the serious global threat of increasing antimicrobial resistance. Microbial activity and biofilms on surfaces cost UK industry billions of pounds each year due to product contamination, energy losses and equipment damage. Infection control, via advanced anti-microbial surfaces, is a key strategy to combat resistance. Uniquely, the BioAFM will enable us to learn how harmful bacteria attach to and respond to surfaces, and develop materials engineered at the nanoscale that can combat infection.2. Developing water-efficient biofuel cropsThere is an urgent and pressing need to improve the ability of biofuel plants to grow productively and sustainably on marginal land that is unsuitable for major food crops. Using the new microscope, we will better understand the mechanisms used by drought-adapted desert plants to conserve water by opening their stomatal pores at night and closing them during the hot, dry light period. These principles will then be applied to generate biofuel crops with these properties. 3. Repair of joints during ageingRegeneration of cartilage, which acts as a flexible cushion between joints, depends on the ability of chondrocytes to produce and maintain the cartilaginous material. Failure in chondrocyte cell function is observed in old age and is associated with osteoarthritis, the most common type of arthritis in the UK, which afflicts around 1 million people every year. With the new microscope, we will follow for the first time, the effects of ageing on the ability of chondrocytes to respond to compression forces at the molecular level and begin to dissect the key pathways that trigger cartilage production.