A Dragonfly multimodal fast imaging platform with SRRF-stream (Super-Resolution Radial Fluctuation) in the Liverpool Centre for Cell Imaging (CCI)

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Individual cells in a plant or an animal are exposed to changes in their environment (biochemical signals, temperature, mechanical forces, light variations...). Cells have to interpret this information to adapt and respond appropriately. However, cells in an organism do not function in isolation, but are part of a complex 3D environment. The ability to experimentally recreate such environments and visualise individual cells in intact organs or 3D cultures is therefore essential to study biological processes. Imaging technologies are developing fast to study individual cells in a multicellular environment 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 microscope (Dragonfly, commercialised by Andor) which allows:1) Fast imaging of the biological processes in real-time, 2) High quality imaging of 3D samples, fixed or living, without the need of specific sample preparation,3) Imaging at very small scales, beyond the optical resolution of traditional microscopes, without damaging the sample due to high levels of light4) Imaging in the near-infrared spectrum of light, which is more commonly used for in vivo imaging. This will enable to integrate measurements done with the same fluorescent probes from cells to tissues in small animal models.5) Imaging in physiological conditions (controlled temperature, humidity, CO2, O2).Dragonfly is a versatile microscope, thanks to its 3 different modes of illumination and is an ideal instrument for a multi-user facility. We will install it in the Liverpool Centre for Cell Imaging (CCI), an open access and shared facility with ~100 registered users from academia and industry. The microscope will serve a breadth of science across the BBSRC remit. We briefly present below two research topics, which will benefit from it:1. A better understanding of photosynthesis in bacteria and algae 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 Dragonfly for fast, high-throughput and high-resolution imaging of the necessary components.2. The unexpected role of oxygen sensing proteins in cell divisionOxygen is essential for life in multicellular organisms and animals have evolved mechanisms to cope with decreased oxygen concentration. Interestingly, oxygen-sensing proteins have recently been discovered, to also regulate essential processes during cell division. The molecular mechanisms that connect oxygen-sensing enzymes and cell division needs to be further investigated, using models of human tissues. Because cell division processes are fast and occur in defined areas of the cells, a microscope allowing high speed imaging and high resolution is required. Moreover, the pseudo-human tissues recreated in vitro for this study are fragile and require the gentle imaging conditions, with low light levels, to avoid light-induced damages. The Dragonfly will provide the required imaging conditions for this study and will enable the discovery of important mechanisms that control cell division in human tissues.