A single cell, single molecule microscopy platform for antibiotics research

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The use of antibiotics to supress and treat bacterial infections is a cornerstone of modern medicine. However, bacteria are increasingly developing resistance towards multiple classes of antibiotics currently in clinical use. Some strains of life-threatening bacteria like tuberculosis are already resistant to all available antibiotics, making them effectively untreatable. The rapidly unfolding antibiotic resistance crisis has been declared a global health emergency by the World Health Organization, and research aiming to tackle antibiotic resistance a strategic priority both by UK Government and UKRI. To address this major threat to human health, we need to ramp up our efforts to screen and develop novel antibiotics that can be used against the multidrug resistant bacteria already in circulation, and also to develop approaches to re-sensitise them for already existing antibiotics. In the longer term, it is crucial to identify novel antibiotic targets and treatment strategies with an intrinsically reduced risk of resistance development. In recent years, bacteria have been found to have a highly complex and dynamic cellular internal organisation. As a result, many of the central properties of antibiotics such as their ability to trigger cell rupture can only be understood in the cellular framework. Following the cellular consequences of a novel antibacterial compound via microscopy is thus an extremely powerful tool to understand how antibiotics work (mode of action). Bacteria also exhibit large cell-to-cell differences across a single population that allows individual bacteria to evade and resist antibiotics. Due to the single cell nature of these phenomena, microscopic techniques are essential for understanding how antibiotics interact with bacterial cells and populations. The Newcastle University Centre for Bacterial Cell Biology (CBCB) is world leading in studying the structure and function of bacterial cells. However, our research focus is not limited to fundamental bacterial cellular biology. Through research on host-pathogen interactions, antibiotic mode of action, identification of novel antibiotic targets, and also through direct novel antibiotic screening projects, researchers at the CBCB are actively engaged in research that aims to translate the gained knowledge to novel antibiotic discoveries and therapies. In the very core of the success of CBCB has been a suite of high performance microscopes dedicated and optimised for work with live bacteria, including pathogenic ones. However, microscopy is still a rapidly developing field with new instrumentation enabling approaches that were previously not feasible. We have identified three complementary, cutting-edge techniques that we foresee to become particularly important for antibiotics research: (i) image-based screening for novel antibiotics, (ii) single cell imaging combined with on-chip drug treatment to understand how antibiotics kill bacteria and how bacteria resist antibiotics, (iii) single molecule microscopy that allows antibiotic action to be monitored directly on the level of individual proteins and complexes. We request funds for the purchase of a microscope system capable of all three techniques, which thus is both extremely powerful and provides excellent value for money. Advanced microscopy systems such as this one require a high level of expertise that often prevents effective adoption of these techniques by non-specialists. To deliver access to these techniques to a widest user base, Newcastle University is supporting this application with commitment to hire a PhD-level staff scientist dedicated to assisting users with experimental planning, image acquisition and image analysis, in addition to managing access and maintenance.