Elyra7 with Lattice SIM microscope in the Liverpool Centre for Cell Imaging (CCI), for fast imaging of living samples beyond the limit of diffraction

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Bacteria or individual cells in a plant or an animal are exposed to changes in their environment (biochemical signals, temperature, mechanical forces, light variations...), and have to interpret this information to adapt and respond appropriately. However, cells in an organism do not function in isolation - they are part of a complex 3D environment. The ability to experimentally recreate such environments and visualise individual cells, and even subcellular organisation in 3D is therefore essential to study biological processes. Imaging technologies are developing fast to study individual cells in a multicellular environment and over-time. The need to visualise cellular processes from the nanometre (to elucidate what happens inside cells) to the millimetre (to elucidate how cells are organised in a tissue) scale in the least invasive manner and in real time has triggered the development of new microscopes and imaging technologies. We propose to purchase a newly commercialised microscope (Elyra 7, from Zeiss), which allows:1) Imaging at very small scales (nanometre), beyond the optical resolution of traditional microscopes, without damaging the sample because of high levels of light;2) Fast imaging of the biological processes in real-time;3) High quality imaging of 3D samples, fixed or living, without the need of specific or complex sample preparation;4) Imaging in physiological conditions (controlled temperature, humidity, CO2, O2).The Elyra 7 is a versatile microscope, with multiple capabilities enabling very fast imaging of fine structures in 3D. We will install it in the Liverpool Centre for Cell Imaging (CCI), an open access and shared facility with ~100 registered users /year, from academia and industry. The microscope will serve a breadth of science across the BBSRC remit. For illustration, we here briefly present below two research topics, which will benefit from it:1. A better understanding of photosynthesis in bacteria to further engineer crops and boost their productivityPhotosynthesis is an essential biological process. During photosynthesis, phototrophs such as cyanobacteria, algae and higher plants convert solar light into chemical energy and generate oxygen necessary for animal life. A better understanding of photosynthesis is required to drive the future engineering of crop plants to increase yields. This is achieved by the elucidation of the organisation of the very efficient photosynthetic machinery in bacteria and in algae. Groups in Liverpool and York have developed molecular tools to visualise it in living bacteria or algae. They now need to be able to image the components very precisely (at nanometre range) and to measure their fast movements. They will use the Elyra 7 for fast, and high-resolution imaging of the necessary components.2. Imaging the processes of ageing in neuronsAgeing leads to alterations in the nervous system, which to various extent impair its functions such as capacity to learn and memory. We need to elucidate the mechanisms causing the alterations in neurons, and determine if they are irreversible or not. We will investigate how ageing affects the organisation of neuronal architecture, the intracellular organelle dynamics and the intracellular protein degradation to understand their contribution to subcellular ageing. We will require very precise imaging to detect fine changes in sub-cellular structures during ageing. We use fruitfly brains as model system, which are organised in 3D, so we will need to not only see the details, but to see them in the entire brain and in real-time. The Elyra 7 with Lattice SIM microscope will allow us to capture subcellular detail and live dynamics in the complexity of the whole brain and with reduced light-induced toxicity, all being necessary to progress our work on the cell biology of neuronal ageing.