Imaging ONEWORLD - 'Using Affimers and finding patterns in super-resolution images' - Professor Michelle Peckham
Scientific Organisers: Stefanie Reichelt, Alex Sossick, Nick Barry, Alessandro Esposito and Kirti Prakash
The meeting will begin at 1pm BST.
As part of the 'Imaging ONEWORLD' series, the focus of these lectures is on microscopy and image analysis methods and how to apply these to your research. Almost all aspects of imaging such as sample preparation, labelling strategies, experimental workflows, ‘how-to’ image and analyse, as well as facilitating collaborations and inspiring new scientific ideas will be covered. Speakers will be available for questions and answers. The organisers, CRUK CI core facility staff, Gurdon Institute, MRC-LMB, MRC Cancer Unit and NPL will be able to continue the discussion and provide advice on your imaging projects.
Professor Michelle Peckham
Executive Honorary Secretary of the RMS
University of Leeds
Michelle is Professor of Cell Biology in the Faculty of Biological Sciences. She obtained a BA in Physiology of Organisms at the University of York, and a PhD in Physiology at University College London. She moved to King's College London, and started to use a specialised form of light microscopy (birefringence) to investigate muscle crossbridge orientation. She then worked at UCSF, San Francisco for a year, where she used fluorescence polarisation to investigate muscle crossbridges. She moved back to the UK, to the University of York, to work on insect flight muscle. In 1990 she was awarded a Royal Society University Research fellowship, based at King's College London, and began working on the cell and molecular biology of muscle development, and started to use live cell imaging to investigate muscle cell behaviour in cultured cells, and confocal microscopy to investigate their cytoskeleton. She collaborated with Graham Dunn to use Digitally Recorded Interference Microscopy with Automatic Phase Shifting (DRIMAPS) to investigate cell crawling behaviour. She moved to Leeds in 1997 as a Lecturer, and has continued to use a wide range of both light and electron microscopy approaches to investigate the molecular motors and the cytoskeleton, most recently using various super-resolution approaches.
Super-resolution microscopy is able to precisely locate specific molecules within cells and cellular structures, to within ~10 nm. Affimers are small (~10kDa, 2-3nm in size) non-antibody binding proteins that can be used instead of antibodies to improve super-resolution microscopy imaging. I will explain what Affimers are, their various uses and their specific application in and advantages for super-resolution imaging. Super-resolution imaging should not just be a pretty picture. We need to find ways to interpret high resolution images in a quantitative way. To achieve this, we have developed a novel method for extracting high-resolution ordered features from localization microscopy data by analysis of relative molecular positions in 2D or 3D. I will explain how this approach allows pattern recognition down to 1% protein detection efficiencies, in large and heterogeneous samples, and in 2D and 3D super-resolution datasets. We have used this method to infer ultrastructure of the nuclear pore, the cardiomyocyte Z-disk, DNA origami structures and the centriole. This versatile tool can be used to find pattern information from samples with unknown structures, and we expect this analysis to be applied to study regularity and symmetry in many other complexes.