High throughput optical mesoscopy for quantitative screening of novel antimicrobial compounds

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Antimicrobial Resistance is one the biggest health challenges facing us today. Research is urgently required to support the rapid discovery and screening of novel antibiotics and to reduce the time and cost involved in bringing these to market. This project seeks to develop key underpinning imaging and analysis techniques to allow effective screening and study of novel antimicrobial candidates, specifically antimicrobial peptides (AMPs) which represent possibly the most promising class of antimicrobial agents. Existing assays such as minimum inhibitory concentration (MIC) suffer from poor reproducibility and cannot provide detailed information about the mode of action of AMPs and the diversity of bacterial responses. Current high resolution optical imaging approaches used at NPL (such as super-resolution microscopy) allow detailed visualisation of only a small population of cells, resulting in low throughput and poor measurement statistics.This project seeks to leverage high throughput imaging and associated image analysis techniques to solve these limitations and provide a powerful new capability for assessing antimicrobials. Prof McConnell has developed a new giant lens called the Mesolens which gives images of hundreds or thousands of mammalian cells in a single image with diffraction-limited resolution. We aim here to extend the capability of the Mesolens to support super-resolution imaging modes using total internal reflection fluorescence (TIRF) microscopy and structured illumination microscopy (SIM). These methods will improve the spatial resolution by a factor of approximately 2 in the lateral direction and 4 in the axial direction to resolve the mode of action of AMPs (such as membrane pore formation) in thousands of cells simultaneously and enable visualisation of AMP action on model membranes. Combining super-resolution mesoscopy with complimentary imaging and biophysical measurements at NPL will provide new insights into the mechanisms of AMP action and allow screening of candidate compounds with unprecedented sensitivity and reliability.