Super-resolution fluorescence atomic force (SURFACE) microscopy

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Bacteria are microscopic organisms of immense importance to humans. Given their diminutive size much of our understanding of how they grow and divide has been determined by a variety of microscopy approaches. Since the earliest microscopes many important discoveries have been made and continued technological developments have permitted new and exciting revelations. Bacterial cell biology and the ability to determine the subcellular localization of components have been largely driven by fluorescence microscopy, whereby individual components are labelled with fluorescent markers. However the limit of detection, coupled with the small dimensions of bacteria has limited the true potential of this approach. Very recently new techniques have allowed fluorescence microscopy to traverse this barrier to attain a level of resolution approaching single molecules. However due to the technical difficulties, lack of commercial availability and expense of some of these developments they have not as yet made a great impact. We have been using a complementary approach called atomic force microscopy, which gives very high resolution of surfaces. We have applied this to the study of bacterial cell walls (the site of action of important antibiotics such as penicillin). This has revealed many important new insights. The aim of the project is to build a new type of microscope coupling super resolution fluorescence and atomic force. We will use our existing microscopy set up as a scaffold to rapidly and efficiently develop the new machine, given the interdisciplinary expertise of the investigator team. The new machine will be tested and further developed within our current bacterial cell wall architecture and dynamics research area. This will provide an ideal framework to develop a new microscope, which can then be utilized and duplicated more widely to allow the rapid uptake of a novel and exciting approach.