10030379Development of the worlds first Live Cell NanoscopeClosedCollaborative R&DInnovate UKLight microscopy has made several key advancements since the invention of the first compound light microscope in the late 16th century. Since then we have moved through several iterations from enhanced magnification, to improved resolution (diffraction limited), targeted fluorescence, high signal to noise confocal and most recently super resolution imaging. As recently as 2014 the Nobel Prize in Chemistry was awarded for 'the development of super-resolved fluorescence microscopy'. In 2015 we launched a revolutionary super resolved fluorescent microscope which not only broke the traditional limits of resolution (Abbe's limit) but also allowed for very fast and gentle imaging, therefore making it the most advanced tool for super resolved 'Live Cell Imaging.' However, this microscope was still limited to what is commonly referred to as the SIM/ISM limit of resolution for a light microscope, which is approximated as Resolution = (0.61\*?)/(2\*NA). So for an emission ? of 525nm and an objective with Numerical Aperture (NA) of 1.49, the best achievable resolution would be 107nm. Whilst this does offer a window into some of the smallest interactions which take place within cell biology, the increase in the amount of information that can be gained from resolving down at the nanoscopy scale (=50nm) is exponential. Currently the only tools available for fluorescence based nanoscopy (SMLM, STED, CLEM) all require the biological specimen to be fixed and therefore do not allow you to see the huge amount of sub cellular interactions which occur during critical biological events such as cell division, protein to protein interaction, cell interactions with viruses and bacteria, cell contraction, etc... What we are proposing to develop as part of this project is the worlds first fluorescent based nanoscope which enables live cell imaging. It should also be noted that the developed instrument would not only be suitable for the scientific research field; advanced light microscopes are no longer confined to scientific laboratories and are being used for a variety of clinical and industrial (pharmaceutical) applications, from medical screening to drug development. Such a high speed, live cell imaging nanoscope would open up new possibilities for screening applications in medical screening such as for the diagnosis of platelet granule disorder, high content screening (HCS) for drug development such as in the development of mRNA vaccines and gene sequencing for high density in-situ sequencing to spatially resolve RNA/DNA/Proteins for development of personalised medicines.