Abstract: The influenza virus is a major cause of morbidity and mortality, and results in a significant health and economic burden worldwide. Despite this, many aspects of the virus are poorly understood. In particular, filamentous forms of the virus are generally overlooked in virus research, despite being highly clinically relevant. In addition, commonly used methods to study virus structure are often low through-put, or lack the resolution required to resolve virus particle features. In order to address this, we have developed a fluorescence super-resolution microscopy and rapid automated analysis pipeline to image many thousands of individual influenza virions at a time, gaining information on their size, morphology and protein distribution. Using this pipeline, we have investigated the size distribution of spherical influenza virions through clustering analysis of super-resolution images, the length distribution of influenza filaments by fitting splines to microscopy images of filaments, and the distribution of viral surface proteins and RNA. We observed broad phenotypic variability in filament size, and Fourier transform analysis of super resolution images demonstrated no generalized common spatial frequency patterning of proteins on the virion surface, suggesting a model of virus particle assembly where the release of progeny filaments from cells occurs in a stochastic way. This work provides novel insights into virus particle structure and organisation and represents a powerful technique that is easily extendable to the study of pleomorphism in other pathogens.

Biography: Nicole Robb is an Assistant Professor at the Warwick Medical School and a visiting lecturer at the University of Oxford. She completed a BSc in Microbiology at Imperial College London before doing a DPhil at the University of Oxford, where she specialised in the field of influenza virology. Following a post-doc in Physics at the University of Oxford she was awarded a Royal Society Dorothy Hodgkin Fellowship to set up her group and moved to the University of Warwick. Her current research interests include super-resolution microscopy, rapid pathogen detection and using biophysical techniques to study viral replication. She holds several patents on viral diagnostic technology and is co-founder of a spinout company that is working on rapid viral testing. She is also PhD Course Director of the University of Warwick’s Institute for Global Pandemic Planning (IGPP), a new interdisciplinary initiative formed to research solutions to mitigate the effects of pandemics.