Super-resolution imaging across the Biosciences

Labelling specific objects in samples such as cells and tissues with a fluorescent dye, and then imaging them using a fluorescent microscope, is a tool that has revolutionised our ability to discover how proteins, DNA, RNA and other cellular structures are organised within cells and tissues. However, until recently, the ability to resolve the fine detail was limited to about 0.25 microns (approximately 300x smaller than the width of a human hair). This is because the amount of fine detail or 'resolution' depends on the wavelength of light and the so-called numerical aperture of the objective lens used to image the sample, two properties that cannot be altered beyond a fixed limit. However, about 20 or so years ago, new approaches to overcome this resolution barrier, collectively known as super-resolution imaging, started to be developed, leading to the award of the Nobel Prize to key developers of this technology (Hell, Moerner and Betzig) in 2014. Super-resolution imaging is rapidly becoming a key tool for researchers to uncover the fine detail of how objects are organised within cells, revealing new detail that we did not previously know existed. In this research, we want to open up a specific type of super-resolution imaging called 'single molecule localisation microscopy' to a wide range of researchers, by implementing a commercial system that is easy to use, multifunctional and can image in much great detail, almost 20x better than we can currently achieve with a standard fluorescence microscope. This new microscope will reveal new insight into protein organisation within cells and tissues across a range of research areas, from cancer to heart disease and blindness.

We have developed several forms of super-resolution microscopy @Leeds through both commercial and homebuilt systems, enabling us to develop key expertise in super-resolution imaging. The BioImaging Facility in the Faculty of Biological Sciences has two commercial Zeiss confocal microscopes with FastAiryscan, that provide a resolutions of about 2 fold better than a standard confocal (~150nm) and a commercial 2D STED microscope that provides a resolution about 5 fold better (~50nm) in X and Y (but with lower resolution in Z). However, many bioscience research areas require even higher resolution to tease out the detailed organisation of proteins within samples, and understand how proteins interact in health and disease. To address this growing need, we plan to implement a commercial single molecule resolution microscope that will allow us to image at even higher resolutions, with 3D isotropic localisation precision (15 x 15 x 15 nm in X,Y and Z). This microscope will provide users with a cost effective, versatile and, importantly, an easy to use approach to this cutting edge technique, opening up new research possibilities across a range of Bioscience, from cancer biology to heart disease.